Recent advances in CAR-MSCs: the new engine of cellular immunotherapy evolution.

Introduction

The progress in chimeric antigen receptor (CAR) design has considerably accelerated the development of cell-based immunotherapies, playing a central role in biomedical studies. Cells engineered with CARs, such as CAR-T and CAR-natural killer (NK) cells, have exhibited impressive outcomes in addressing a range of blood-based and solid cancers [ 1 – 5 ]. The technology itself has advanced through several generations—from first to fifth—each aimed at improving treatment effectiveness and reducing risks [ 6 , 7 ]. Nevertheless, issues like immune reactions, adverse effects, and short-lasting responses remain substantial barriers [ 7 ]. These limitations have encouraged the exploration of new cell types to augment the capabilities of CAR-based treatments.

Mesenchymal stem cells (MSCs), constituting a type of multipotent stromal cell, can be sourced from diverse tissues such as bone marrow (BM), adipose tissue, and the umbilical cord (UC), followed by extensive in vitro expansion [ 8 ]. These cells are characterized by their low immunogenicity, capacity to facilitate tissue repair, and potent immunomodulatory functions within the microenvironment [ 9 ]. Their broad therapeutic applicability and an excellent safety profile are exemplified in over 1,300 clinical trials conducted globally, notably for conditions like graft-versus-host disease (GvHD) and osteoarthritis [ 10 , 11 ].

By integrating CAR technology with MSCs, researchers have developed CAR-MSCs, which combine the targeting specificity of CARs with the inherent advantages of MSCs. This fusion positions CAR-MSCs as a novel frontier in cell-based therapies [ 12 – 14 ]. CAR-MSCs can be engineered to target tumor-associated antigens while simultaneously modulating the tumor microenvironment (TME), potentially mitigating the systemic toxicity. Moreover, the integration of CAR technology with MSCs expands the spectrum of potential targets for cancer treatment and opens new avenues for applying CAR technology in the treatment of nonmalignant conditions, such as immune-mediated disorders, neurological diseases, and tissue repair and regeneration [ 15 – 17 ].

The development of CAR-MSCs marks a significant step forward in cellular immunotherapy, offering more effective and safer treatment strategies. This review systematically summarizes the evolution of CAR engineering in conjunction with the biological characteristics and therapeutic applications of MSCs. For the first time, we comprehensively discuss the engineering design, biological properties, and clinical applications of CAR-MSCs in the treatment of tumors and immune-related diseases. Furthermore, we also discussed the potential challenges and corresponding strategies that the future clinical application of CAR-MSCs might encounter. Through a critical evaluation of the literature, our aim is to highlight the significant potential of CAR-MSCs in the field of immunotherapy.

Current status of CAR engineering

The CAR is a complex synthetic receptor with a history spanning nearly three decades [ 18 ]. As a specific receptor, CAR enables immune cells to recognize antigens and elicit antigen-specific cytotoxic responses. The significant progress in immune cell therapies offers robust evidence supporting the strategic decision made decades ago to integrate CAR technology into T cells [ 19 ].

The CAR structure includes an extracellular domain composed of a hinge region and a single-chain variable fragment (scFv). The scFv, derived from antibody variable regions, binds tumor cell surface antigens independently of major histocompatibility complex (MHC). The hinge region connects the scFv to the transmembrane domain. The intracellular part of CAR contains signaling domains from T-cell receptors and costimulatory molecules. These domains are typically derived from CD28 family receptors or TNF receptor family members (e.g., 4-1BB). Together, they boost T-cell expansion and cytokine production by activating costimulatory pathways, enhancing the antitumor activity of T cells [ 20 – 22 ]. Signal transduction domains usually include the TCR/CD3ζ or FcεRIγ chain, both containing immunoreceptor tyrosine-based activation motif essential for T-cell signaling. CARs are capable of autonomously recognizing tumor-associated antigens (TAAs) without restriction by the MHC. These engineered receptors are designed to increase the ability of the immune system to recognize and eliminate cancer cells [ 23 , 24 ].

Since the 1980 s, CAR technology has undergone multiple iterations of refinement, significantly increasing the breadth of cellular immunotherapy (Fig. 1 ). The advancements in cellular immunotherapy centered on CARs exemplify the translation of fundamental research into clinical applications. First-generation CAR-T cells contain only a single intracellular signaling domain, such as CD3ζ or FεRIγ, without any costimulatory domain. These CAR-T cells resemble endogenous TCRs; however, their limited ability to generate interleukin-2 (IL-2) results in a diminished response during clinical trials. Consequently, supplementation with exogenous IL-2 is often necessary for first-generation CAR-T cells to ensure an efficacious response [ 25 ].

Fig. 1 Architecture of CARs and their successive iterations. First-generation CARs contain a single activation domain, such as CD3ζ, which triggers cell activation and scFv-specific cytotoxicity. Second- and third-generation CARs include one or two costimulatory signals, enhancing both costimulatory molecule activation and intracellular signaling. Fourth-generation CARs, or TRUCKs, combine previous features to target and eliminate tumor cells while simultaneously secreting cytokines like IL-12 through cytokine gene cassettes, boosting the local immune response. Abbreviations: CAR, chimeric antigen receptor; TNF, tumor necrosis factor; TRUCK, T cell redirected for universal cytokine-mediated killing

The limitations of first-generation CARs were addressed in second-generation CAR-T cells by incorporating costimulatory signaling domains (e.g., CD28 or 4-1BB). Studies have demonstrated that engineered costimulatory domains (e.g., ICOS and CD27) exhibit significant antitumor effects [ 21 , 22 ]. These enhancements improve T-cell proliferation, cytokine production, cytotoxicity, and in vivo persistence. The effects of different costimulatory domains vary to some extent. Compared with CD28, the 4-1BB costimulatory domain exhibits longer persistence and reduced cell exhaustion [ 26 ]. These advancements have substantially improved the therapeutic efficacy of CAR-T cells. Currently, most FDA-approved CAR-T-cell therapies utilize the second-generation CAR design.

Third-generation CARs incorporate two costimulatory signaling domains in conjunction with the CD3ζ chain [ 27 ]. This design aims to optimize T-cell activation, proliferation, persistence, and antitumor functionality via dual costimulatory signaling. However, preclinical studies of third-generation CAR-T cells have yielded inconsistent results; some investigations have demonstrated superior performance compared with that of second-generation CAR-T cells, whereas others have reported no significant difference in efficacy between the two [ 28 , 29 ]. The reasons for the disparate outcomes observed in third-generation CAR-T-cell therapies remain unclear and may be attributed to factors such as limited sample sizes, potential instability in product manufacturing, or variations in the immune microenvironment. Third-generation CAR-T-cell therapy targeting CD19 has been reported to be an effective and safe treatment for systemic IgM light chain amyloidosis (AL) associated with underlying marginal zone lymphoma [ 30 ]. However, the enhanced efficacy from dual costimulatory signals may also increase the risk of adverse events, especially severe cytokine release syndrome (CRS). Therefore, the selection and optimization of costimulatory molecules are crucial for preventing excessive signaling, which may lead to immune cell exhaustion. More clinical trials are needed to evaluate and monitor the efficacy and safety of these products, helping to unlock the full potential of third-generation CARs.

Fourth-generation CARs, or TRUCKs (T cells redirected for universal cytokine killing), recognize tumor cells and induce cytotoxicity through CARs while also releasing engineered immunomodulators (e.g., IL-12, IL-15, and IL-2) to modulate the TME. For example, to combat T-cell dysfunction, Zhao Y et al. engineered CAR-T cells to secrete IL-10 [ 31 ]. This modification improved CAR-T cell proliferation and effector functions, leading to complete regression of multiple cancers, such as colon cancer, breast cancer, melanoma, and pancreatic cancer, in mouse models. Moreover, TRUCKs can enhance CAR-T-cell functionality by expressing chemokine receptors or chemokines (e.g., CCL19 and CCL21), enabling directional migration to the TME [ 32 ]. These strategies have been validated in preclinical studies and shown promising efficacy and safety in clinical trials for various malignant tumors [ 33 , 34 ]. The ability to elicit a secondary antitumor immune response distinctly endows fourth-generation CARs with significant therapeutic potential for treating solid tumors, particularly in targeting antigen-negative cancer cells within the TME [ 35 ].

Furthermore, similar strategies involve engineering CAR-T cells with gene-edited noncytokine proteins, which can be categorized into secretory and nonsecretory types. Secretory proteins (such as Helicobacter pylori neutrophil-activating protein, drug-activating enzymes, scFv-based PD-1 blockers, and heparinase) typically target the immunosuppressive TME [ 36 – 38 ]. Nonsecretory proteins (such as c-Jun, PD-L1-specific chimeric conversion receptors, anti-B7-H3 chimeric cytokine receptors, anti-CD38 chimeric cytokine receptors, and TGFβR 4-1BB hybrid receptors) primarily enhance the functionality of CAR-T cells [ 39 – 42 ]. Both approaches have demonstrated potential in preclinical studies for treating hematological malignancies and solid tumors. Consequently, fourth-generation CAR-T-cell therapy represents a highly promising therapeutic strategy. In the future, it will be essential to further investigate and optimize the structure of TRUCKs and evaluate their efficacy through additional preclinical and clinical trials to achieve more efficient tumor cell clearance.

Research on fifth-generation CARs remains in the exploratory phase, with mainstream investigations and clinical applications focused primarily on the first four generations of CARs. Currently, fourth-generation CARs are undergoing active research and early clinical trials. The term ‘fifth generation’ may refer to further enhancements and innovative therapies derived from the fourth generation. Some scholars have reported the development of fifth-generation CARs by modifying second-generation CARs with the addition of the IL-2Rβ cytokine receptor in the intracellular domain. This change enables coordinated activation of the JAK-STAT signaling pathway between costimulatory signals (e.g., 4-1BB or CD28) and CD3ζ, promoting improved cell proliferation and persistence [ 43 ]. To enhance CAR-T cell efficiency, Yuti P et al. developed fifth-generation anti-BCMA CAR-T cells containing three costimulatory domains and capable of secreting anti-PD-L1 scFv blockade molecules. These cells demonstrated superior antitumor efficiency, enhanced proliferative capacity, and reduced T-cell exhaustion in response to multiple myeloma (MM) cells [ 44 ]. However, there is no widely accepted definition of the terminology for the fifth generation within the academic community. In summary, the available data suggest that the overall efficacy and safety profile of CAR-T-cell therapy is highly encouraging [ 45 , 46 ].

However, the comprehensive advancement of CAR-T-cell therapy has been significantly impeded by several challenges, including limited tissue penetration, antigenic evasion, tumor heterogeneity, an immunosuppressive TME, increased treatment expenses, immune effector cell-associated neurotoxicity syndrome (ICANS), and CRS [ 47 , 48 ]. Consequently, the industry has acknowledged that the full potential of CAR technology should not be confined solely to T-cell modification; it is equally imperative to leverage the antitumor properties of other immune cell types, including NK cells, macrophages, invariant natural killer T (iNKT) cells, γδT cells, regulatory T cells (Tregs) and MSCs [ 15 , 49 – 51 ]. In addition, evidence from both clinical and preclinical studies has built upon the success of CAR-T-cell therapy in oncology to extend its applications to various noncancerous diseases, including autoimmune disorders (e.g., systemic lupus erythematosus (SLE) and inflammatory myopathy), fibrotic conditions (e.g., cardiac disease, liver disease, chronic kidney disease, pulmonary disorders, and skeletal muscle diseases), and senescence-associated pathologies (e.g., liver fibrosis, solid tumors, atherosclerosis, and natural aging). Infectious pathologies such as HIV, hepatitis B/C, cytomegalovirus, and tuberculosis—initial targets of CAR therapy—are being re-evaluated in light of a deeper understanding of immune responses and the advent of enhanced therapeutic tools [ 52 ]. Compared with tumors, noncancerous diseases typically present a significantly smaller target burden, where partial clearance of diseased cells can yield therapeutic benefits and present a lower mutational load. In addition, the immunosuppressive environment characteristic of the TME is largely absent in most chronic diseases, allowing physiological targeting of diseased tissue by the immune system [ 52 ].

Biological properties of MSCs

MSCs represent a heterogeneous population of multipotent adult stem cells with the ability to self-renew and differentiate [ 53 ]. Initially identified as colony-forming unit fibroblasts in the 1970 s, MSCs were discovered within the BM and are morphologically defined as spindle-shaped, adherent nonhematopoietic stem cells (non-HSCs) [ 54 , 55 ]. Besides BM, these cells can be obtained from various tissues and organs such as adipose tissue, UC tissue, Wharton’s jelly, amniotic fluid, placenta, liver, heart, lung, adrenal glands, pancreas, and dental tissues. MSCs can also be derived from embryonic stem cells or induced pluripotent stem cells (iPSCs) (Fig. 2 ) [ 8 , 56 – 61 ]. To better define human MSCs, the Mesenchymal and Tissue Stem Cell Committee of the International Society for Cell Therapy (ISCT) has proposed that MSCs should express CD73, CD90, and CD105 in over 95% of cases, while lacking CD45, CD34, CD14 or CD11b, CD79a or CD19, and human leukocyte antigen DR (HLA-DR) surface molecules in less than 5% [ 62 ]. Under specific conditions, MSCs can differentiate into key cell lineages, including osteoblasts, chondrocytes, myocytes, and adipocytes. The differentiation direction, differentiation potential, and heterogeneity of MSCs vary according to their tissue origin. For example, MSCs derived from the BM, synovium, and periosteum present the highest alizarin red positivity rate for osteogenesis; amniotic MSCs demonstrate superior chondrogenic potential [ 63 ]; the number of BM-MSCs decreases sharply with age. Moreover, fetal MSCs possess greater proliferation capacity [ 64 ].

Emerging evidence has expanded the surface marker repertoire of MSCs, complementing well-established canonical markers with novel biomarkers, including SERPINF1, S100A9, LRRC75A, CMKLR1 (CCRL2), CD142, CD271 (LNGFR), TNFAIP6 (TSG6), CSPG4, CD106 (VCAM-1), CD317, and CD146 [ 65 ]. These refined surface marker signatures enable the stratification of functionally distinct MSC subpopulations, particularly those orchestrating niche-specific functions in HSC maintenance, neoangiogenesis, neurogenic niche modulation, and immune regulation, as evidenced by recent single-cell RNA sequencing and secretome profiling studies (Table 1 ). In a myocardial infarction model, CD73 + adipose-derived (AD) MSCs demonstrated superior cardiac recovery through enhanced angiogenesis. Compared with their CD73-negative counterparts, these cells showed significantly increased secretion of vascular endothelial growth factor (VEGF), stromal cell-derived factor-1α (SDF-1α), and hepatocyte growth factor (HGF) [ 66 ]. CD271 + AD-MSCs exhibited elevated angiogenic gene expression with concurrent downregulation of inflammatory genes, maintaining characteristic MSC surface markers while demonstrating enhanced neoangiogenic potential [ 67 ]. TNFAIP6 + murine MSCs displayed increased immunosuppressive activity and therapeutic efficacy in acute inflammation models [ 68 ]. Neuroglial antigen 2 (NG2)-expressing BM-MSCs presented increased proliferative capacity, preserved stemness, and improved functional recovery in hepatic fibrosis models [ 69 ]. CD106 + MSCs from UC sources demonstrated superior adhesive properties, angiogenic capacity, wound-healing potential, and immunomodulatory functions [ 70 ]. CD317 + MSCs exhibited increased differentiation potential alongside increased immunosuppressive activity [ 71 ]. CD146 + cell populations have greater cartilage-protective effects and Th17 cell suppression capabilities [ 72 ].

Table 1 Functional properties of biomarker-defined MSC subpopulations in preclinical models Surface Marker MSC Source Disease Model Functional Outcomes Mechanistic Insights Ref. CD73 Mice AD MI in rats Superior cardiac recovery via enhanced angiogenesis Upregulated VEGF, SDF-1α, and HGF secretion [ 66 ] CD271 Human AD Tissue regeneration Improved neoangiogenesis with an anti-inflammatory profile Elevated angiogenic gene expression with downregulated inflammatory genes; retains typical stem cell surface marker complement [ 67 ] TNFAIP6 Mice BM, placenta, and AD Acute inflammation in mice Augmented immunosuppressive activity Accelerated anti-inflammatory cytokine TNFAIP6 release [ 68 ] NG2 Mice BM Liver fibrotic/cirrhotic injury in mice Marked motion and proliferation; stronger inhibition of inflammatory infiltration and fibrosis; better balance of immune response Greater stemness potential with increasing Ki-67/SSEA-3 expression [ 69 ] CD106 Human UC Full-thickness wound mouse model Superior adhesive properties, angiogenic capacity, wound healing, and immunomodulation / [ 70 ] CD317 Human AD Acute inflammation in mice Enhanced differentiation potential and immunosuppressive activity Higher levels of TNFAIP secretion [ 71 ] CD146 Human UC Collagen-induced arthritis in mice Greater cartilage-protective effects More conspicuous Th17 cell inhibition resulting in a lower level of IL-17 A [ 72 ] MSCs, mesenchymal stem cells; AD, adipose-derived; BM, bone marrow; UC, umbilical cord; MI, myocardial infarction; VEGF, vascular endothelial growth factor; SDF-1α, stromal cell-derived factor-1α; HGF, hepatocyte growth factor; NG2, neuroglial antigen 2; SSEA-3: stage-specific embryonic antigen-3; Th17, helper 17; IL-17 A, interleukin-17 A; TNFAIP6, tumor necrosis factor alpha-induced protein 6; Ref., reference

Building on our previous research [ 73 – 76 ], we employed single-cell sequencing to delineate MSC heterogeneity. A CDCP1-expressing MSC subgroup showed enhanced migration and superior acute GvHD suppression compared to its CDCP1-negative counterpart. CDCP1, designated CD318, functions as a transmembrane glycoprotein predominantly expressed in tissue-specific progenitor populations, including HSCs, neural progenitor cells (NPCs), and MSCs [ 77 ]. Mechanistically, we hypothesized that CDCP1 enhances the homing of MSCs to GvHD-affected tissues via RhoA pathway activation, amplifying their immunomodulatory effects. Current animal studies are validating these properties to optimize the selection of clinical-grade MSCs for acute GvHD prophylaxis. In conclusion, the functional heterogeneity of MSCs is closely associated with their surface marker profiles. Moving forward, leveraging single-cell transcriptomics and proteomics will be key to precisely identifying, functionally enhancing through genetic engineering, and advanced sorting of MSC subpopulations, ultimately enabling the isolation of disease-specific MSCs. On this basis, CAR-engineered MSCs hold promise as potent tools for personalized medicine.

Bidirectional interactions between MSCs and CAR-engineered cells

Emerging evidence has revealed bidirectional interactions between MSCs and CAR-engineered immune effectors, particularly CAR-T cells (Table 2 ). The current literature reveals three distinct paradigms: MSC-mediated suppression of CAR-T-cell functionality, enhancement of CAR-T-cell activity, and neutral effects on cellular potency.

Table 2 Bidirectional interactions between MSCs and CAR-engineered cells: Outcomes, mechanisms, and therapeutic implications Interaction Type Key Findings Mechanism Model System Potential Clinical Application Ref. Suppression of CAR-T-Cell Activity BM-MSCs inhibit AML-targeted CAR-T-cell expansion via IDO-1 upregulation; reduce IFNγ/IL-2 secretion IDO-1, COX-2, PD-L1, PD-L2, and ICAM-1 mediated immunosuppression Cell coculture IDO-1 inhibitors enhance CAR-T-cell expansion [ 78 ] hUC blood-MSC-derived STC1 suppresses the efficacy of CAR-T cells in killing lymphoma cells STC1-induced TME modulation Cell coculture; xenograft mice STC1 blockade to enhance CAR-T efficacy [ 79 ] BM-MSCs protect MM cells against low-affinity, moderate-lytic CAR-T via cell-contact dependency CAR affinity-dependent resistance Cell coculture Optimizing CAR affinity/design for MM [ 80 ] BM-MSCs induce resistance to NK cells in MM; CD38-CAR engineering reverses protection CAR-dependent evasion of MSC-mediated resistance Cell coculture CAR engineering to bypass MSC-mediated resistance [ 81 ] Enhancement of CAR-T-Cell Function UC-MSC mitochondrial transfer reduces CAR-T-cell apoptosis and enhances cytotoxicity Increased mitochondrial activity of anti-apoptotic Bcl-2/BARD1 pathways Cell coculture MSC mitochondrial transfer potentiates the therapeutic effects of virus-free generated CAR-T cells [ 82 ] BM-MSCs with OAd/IL-12/PD-L1 inhibitors enhance HER.2 CAR-T-cell-mediated tumor killing in lung tumors Combination therapy boosted T-cell effector cytokines (IFN-γ, granzyme B, perforin) 3D tumor spheroids; NSCLC xenograft mice MSC-vectored combination therapies [ 83 ] IL-7/IL-12-secreting BM-MSCs boost CAR-T-cell expansion and Th1 polarization IL12 improved CAR-T cell proliferation, reduced AICD, and enhanced target cell lysis Colorectal cancer xenograft mice Genetically engineered MSCs for CAR-T adjuvants [ 84 ] Neutral Effects on CAR-T Cells BM-MSCs derived from B-ALL pediatric patients suppress conventional T cells but spare CD19-CAR-T-cell cytotoxicity CAR-T-cell insensitivity to B-ALL BM-MSC-mediated immunosuppression Cell coculture; acute colitis mice and B-ALL xenograft mice Decoding BMM-driven CAR-T-cell resistance mechanisms [ 85 ] MSCs promote hematopoietic recovery without impairing CAR-T-cell antitumor efficacy CXCL12/NF-E2-mediated BM reconstitution B-cell lymphoma mice MSCs for post-CAR-T-cell hematopoietic support [ 86 ] BM-MSCs, bone marrow-derived mesenchymal stromal cells; hUC blood-MSCs, human umbilical cord blood-derived mesenchymal stromal cells; UC-MSCs, umbilical cord-derived mesenchymal stromal cells; TME, tumor microenvironment; MM, multiple myeloma; OAd, oncolytic adenovirus; CAR-T, chimeric antigen receptor T cell; AML, acute myeloid leukemia; IDO-1, indoleamine 2,3-dioxygenase 1; COX-2, cyclooxygenase 2; PD-L1, programmed death-ligand 1; ICAM-1, intercellular adhesion molecule 1; IFNγ, interferon-gamma; IL, interleukin; STC1, stanniocalcin-1; NK, natural killer; CD, cluster of differentiation; Bcl-2, B-cell lymphoma 2; BARD1, BRCA1-associated RING domain protein 1; NSCLC, non-small cell lung cancer; AICD, activation induced cell death; B-ALL, B-cell acute lymphoblastic leukemia; BMM, bone marrow microenvironment; BCMA, B-cell maturation antigen; Th1, T helper 1 cell; CXCL12, C-X-C motif chemokine ligand 12; NF-E2, nuclear factor erythroid-derived 2; B-ALL, B-cell acute lymphoblastic leukemia; Ref., reference

MSC-mediated suppression of CAR-T-cell efficacy

Towers et al. demonstrated that BM-MSCs suppress acute myeloid leukemia (AML)-redirected switchable CAR-T-cell expansion through IDO-1-mediated mechanisms, concurrently attenuating IFNγ and IL-2 secretion [ 78 ]. Zhang et al. identified stanniocalcin-1 (STC1) as a critical MSC-secreted factor that modulates the TME to impair CAR-T-cell cytotoxicity against lymphoma in murine xenograft models [ 79 ]. Holthof and colleagues employed a diverse set of CAR-T cells exhibiting a range of antigen specificities—including BCMA, CD38, and CD138—along with varying affinities for target cells [ 80 ]. The results showed that MM patient-derived BM-MSCs significantly protected MM cells against lower-affinity, moderately lytic CAR-T cells. Similarly, MSCs protected MM cells against KHYG-1 NK cells by inducing resistance mechanisms. However, the immune resistance mediated by MSCs can be reduced by genetically modifying KHYG-1 cells to express the CD38 CAR [ 81 ]. These findings suggest that this immune escape can be addressed by inhibiting antiapoptotic molecules or enhancing immune killer cells.

MSC-mediated enhancement of CAR-T-cell function

Recently, Court et al. demonstrated that mitochondrial transfer from MSCs reduced CAR-T-cell apoptosis postelectroporation while enhancing cytotoxic activity, suggesting metabolic reprogramming as a mechanism for improved stress resistance [ 82 ]. McKenna et al. engineered MSCs to deliver oncolytic adenoviruses (OAds) coexpressing IL-12 and PD-L1 blockade, ​which enhanced​ CAR-T-cell antitumor activity against solid tumors by combining localized viral oncolysis with immune checkpoint inhibition [ 83 ]. Hombach et al. pioneered the secretion of IL-7/IL-12 by transgenic MSCs, which potentiated CAR-T-cell expansion in colorectal cancer models, achieving superior tumor control compared with unmodified MSC coadministration [ 84 ].

Effects of neutral MSCs on CAR-T-cell activity

Certain studies have demonstrated that MSCs mediate immunomodulation without direct interference with CAR-T-cell functionality. Zanetti et al. characterized BM-MSCs from pediatric B-ALL patients and confirmed their immunosuppressive effects on conventional T cells while preserving CD19-CAR-T-cell cytotoxicity [ 85 ]. Xia et al. identified key mechanisms by which MSCs support hematopoietic recovery following CAR-T-cell therapy. Notably, the murine model demonstrated that coadministration of MSCs does not impair CAR-T-cell antitumor efficacy [ 86 ].

Applications of MSCs

MSCs possess significant potential for clinical applications because of their unique therapeutic properties, which include low immunogenicity, inherent tumor tropism, ease of isolation, regenerative capabilities, and multidirectional differentiation potential [ 9 ]. Through direct cell contact and the release of cytokines, chemokines, and growth factors, MSCs help sustain immune balance across most tissues by influencing inflammation, blood vessel formation, and tissue fibrosis (Fig. 2 ) [ 87 ]. The distinctive characteristics and mechanisms offer exceptional therapeutic prospects, which has prompted the use of MSCs as a cell therapy technology in most research worldwide [ 88 ]. As of February 2025, ClinicalTrials.gov hosts over 1300 projects focused on MSCs.

Immune disorders

MSCs block the T-cell cycle, promote Th2/Treg polarization, inhibit B-cell proliferation and antibody secretion, suppress NK-cell proliferation, and drive macrophage polarization from the M1 phenotype to the M2 phenotype, thereby reducing inflammation, prolonging neutrophil survival, and inhibiting DC differentiation [ 89 – 91 ].

Hematopoietic stem cell transplantation (HSCT) holds an irreplaceable position in treating malignant hematological diseases [ 92 , 93 ], as it offers patients the possibility of a cure by eliminating malignant cells and reconstructing the hematopoietic and immune systems. However, GvHD is a major complication after HSCT that seriously threatens patient survival and quality of life [ 94 ]. MSCs show strong effectiveness in treating or preventing GvHD, with clinical studies confirming their ability to improve patient survival and quality of life [ 10 , 95 – 98 ]. Moreover, across multiple trials targeting immune-related conditions, most also demonstrated both safety and efficacy [ 99 – 102 ]. Notably, a recent study demonstrated that CAR-modified MSCs can significantly enhance the immunosuppressive effects at inflammatory sites of colon GvHD in murine models, mitigate the severity of GvHD, prolong survival duration, and concurrently preserve both the stem cell phenotype and safety profile of MSCs [ 103 ]. This discovery signifies a transformative advancement in the application of cell therapy for immune-related disorders, offering robust conceptual validation for designs and enhancements of CAR-MSCs.

Fig. 2 The origins and therapeutic biological properties of MSCs. Initially discovered in BM, MSCs have subsequently been obtained from a range of other biological sources such as adipose tissue, UC, dental pulp, amniotic fluid, and the placenta. These cells release a wide variety of biologically active substances—including growth factors, cytokines, and extracellular vesicles—that contribute significantly to processes like immune modulation, tissue regeneration, blood vessel formation, and neural protection. Abbreviations: MSC, mesenchymal stem cell; BM, bone marrow; UC, umbilical cord; TGF, transforming growth factor; IL, interleukin; CCL, chemokine (C-C motif) ligand; miR, microRNA; HGF, hepatocyte growth factor; VEGF, vascular endothelial growth factor; KGF, keratinocyte growth factor; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor; IGF, insulin-like growth factor; OPG, osteoprotegerin; BDNF, brain-derived neurotrophic factor; SDF, stromal cell-derived factor

Hematological disorders

Researchers have demonstrated that the combination of MSCs and HSCs can increase transplantation success rates [ 104 – 106 ]. Mechanistically, MSCs may promote the expansion of HSCs, thereby enhancing hematopoietic recovery after CAR-T-cell therapy through the upregulation of C-X-C motif chemokine ligand 12 and nuclear factor erythroid derived 2 [ 86 ]. The AKT signaling pathway—a key modulator of MSC biological activity—has also been shown to govern HSC proliferation and differentiation. MSC-derived microvesicles, which carry hematopoietic niche-associated mRNAs that enhance hematopoietic reconstitution through paracrine reprogramming, have been identified as critical mediators of HSC engraftment. Furthermore, MSCs suppress donor lymphocyte-mediated cytotoxicity and regulate post-transplantation inflammatory responses, thereby enhancing engraftment efficiency in allogeneic transplantation models [ 104 , 107 ]. Some studies have also preliminarily investigated the potential therapeutic benefits of MSCs in aplastic anemia and immune thrombocytopenia [ 108 , 109 ].

Tissue regeneration and repair

MSCs possess robust tissue repair capabilities and are utilized to address various types of tissue injuries, including those affecting bone [ 110 ] and cartilage [ 111 ], tendinopathy [ 112 ], and those affecting the liver [ 113 ] and the spinal cord [ 114 ]. Current trials have shown that MSCs are safe and effective for alleviating pain and enhancing cartilage thickness in osteoarthritis patients [ 11 ]. However, they have not been shown to significantly improve quality of life [ 115 ]. Additionally, MSCs show promise in cardiac regeneration by promoting cardiomyocyte repair and improving heart function [ 116 ]. Intracoronary infusion of MSCs after acute myocardial infarction has been proven to improve the left ventricular ejection fraction [ 117 ]. Although direct intramyocardial injection of MSCs is safe for treating chronic heart failure, it is ineffective in restoring cardiac function or alleviating symptoms [ 118 ]. The application of MSCs for skin wounds, limb ischemia, and tissue regeneration is expanding, with increasing evidence supporting their role in promoting angiogenesis and facilitating cell repair [ 119 , 120 ].

Neurological diseases

MSCs have shown potential for enhancing neurological repair functions, particularly in conditions like Parkinson’s and Alzheimer’s diseases [ 121 , 122 ]. They support neural regeneration by differentiating into neural cells and secreting neurotrophic factors that enhance nervous system function [ 123 ]. Moreover, MSCs have demonstrated beneficial effects in treating stroke (including ischemic stroke and acute cerebral hemorrhage), chronic motor deficits, and optic neuropathy, broadening their therapeutic potential [ 124 – 128 ]. Overall, clinical trials have confirmed the efficacy, feasibility, and safety of MSCs, laying the groundwork for future research to benefit more patients.

Cancer

The mechanisms by which MSCs exert their effects in cancer therapy are intricate and multifaceted [ 9 ]. The key mechanisms include the following: [ 1 ] Immunomodulatory capacity: MSCs modulate the immune components of the TME through direct cell–cell interactions and paracrine signaling via soluble factors like TNF-α and IFN-γ, inducing cancer cell apoptosis and reducing invasiveness [ 129 , 130 ] [ 2 ]. Induction of cancer cell apoptosis: Studies show that coculturing MSCs with specific cancer cell types induces apoptosis and inhibits proliferation. MSCs induce apoptosis in breast cancer cells in a dose-dependent manner [ 131 ], and BM-MSCs significantly trigger both early and late apoptosis in CD34 + leukemic stem cells [ 132 ] [ 3 ]. Homing effect and tumor tropism: MSCs exhibit homing and tumor tropism, migrating to primary tumors, metastatic sites, and areas of tissue damage or inflammation. This property highlights their potential as a targeted delivery system for anticancer agents. Engineered MSCs can produce higher levels of anticancer proteins at tumor sites, effectively inhibiting tumor growth and metastasis [ 133 ] [ 4 ]. Increased effectiveness of radiation therapy and chemotherapy: BM-MSCs, in conjunction with low-dose radiation, can significantly increase the proliferation of peritumoral astrocytes and exhibit notable antiangiogenic properties [ 134 ]. BM-derived MSCs pretreated with doxorubicin exhibit significant cytotoxic effects on breast cancer cells and gastric cancer cells [ 135 , 136 ] [ 5 ]. Paracrine mechanisms: MSCs can be engineered to secrete bioactive molecules that regulate tumor-related processes such as proliferation, survival, angiogenesis, and immunosuppression, thereby inhibiting tumor growth and exerting antitumor effects [ 137 , 138 ].

Fewer clinical trials have explored MSCs for cancer treatment compared to other diseases, with only a limited number completed and most still in recruitment or ongoing stages. Orae-Yazdani S et al. first assessed the safety of herpes simplex virus-thymidine kinase gene therapy using MSCs in recurrent glioblastoma (GBM) patients, confirming its safety [ 139 ]. In the future, multiarm clinical trials are needed to compare the efficacy of this MSC-based regimen with that of standard treatment, not only in GBM but also across other tumor types.

Other diseases

MSCs have also been utilized in the treatment of various other conditions, such as type 2 diabetes and its related complications [ 140 – 142 ], COVID-19/ARDS [ 143 , 144 ], idiopathic nephrotic syndrome [ 145 ], chronic low back pain [ 146 ], and radiation-induced hyposalivation [ 147 ], among others. In conclusion, almost all trials have shown that the treatment methods for MSCs are at least safe. As knowledge of MSC mechanisms advances, future studies will focus on their potential use in treating additional diseases.

MSCs as CAR carriers

MSCs are recognized for their capacity to regulate the immune system, combat inflammation, and promote immune tolerance. They continuously produce or overexpress various proteins and exosomes, enabling direct delivery of therapeutic substances or mediating gene transfer. Thus, MSCs hold promise for use in gene delivery and regenerative medicine [ 148 , 149 ]. Although MSCs have been extensively studied for the treatment of immune-related diseases and are generally considered safe, their therapeutic efficacy remains unsatisfactory. This may be attributed to inadequate targeting and tracking accuracy, as well as limited immunosuppressive capacity [ 150 ]. We propose addressing these challenges by optimizing targeting strategies and enhancing immunosuppressive mechanisms.

The CAR is a genetically engineered protein widely utilized in cell therapy. Its primary functions include (1) targeting and binding to specific antigens on the surface of target cells; (2) providing a stable binding interface to ensure sustained interaction between the CAR and the target; and (3) activating signal transduction pathways to trigger robust immune responses. CAR technology has achieved remarkable success in treating hematological malignancies, leading to FDA approval of CAR-T-cell therapy [ 47 ]. The intrinsic advantages of MSCs make them among the most promising candidates for developing CAR carriers. MSCs can be engineered to express CARs as molecular tools for identifying disease-associated markers.

In this way, compared with traditional CAR-T and CAR-NK cell therapies, CAR-MSCs demonstrate unique advantages in targeting precision, safety profiles, and application versatility. First, CAR-MSCs not only preserve the inherent immunomodulatory properties of native MSCs but also enhance the secretion of anti-inflammatory factors through engineered CAR signaling, thereby achieving a dual therapeutic effect of target localization and tissue repair [ 32 , 148 ]. In contrast, CAR-T therapy may trigger CRS due to excessive T-cell activation. Allogeneic CAR-T cells carry the risk of GvHD, as T cells may recognize host antigens [ 48 ]. Although CAR-NK cells are associated with a lower risk of CRS, they may still release inflammatory cytokines [ 151 ]. Notably, MSCs themselves possess immunosuppressive properties that reduce the likelihood of immune rejection, eliminating the need for HLA matching and significantly lowering the risk of GvHD. Second, CAR-T cell therapy involves patient-specific customization, making the process time-intensive and expensive. While CAR-NK cells can be used in an allogeneic setting, their large-scale expansion remains technically challenging. MSCs, on the other hand, are readily available from diverse sources, can be efficiently expanded ex vivo, and maintain stable functionality after cryopreservation—making them highly suitable for mass production as off-the-shelf therapeutic products. Furthermore, CAR-T and CAR-NK cells are predominantly applied in hematological malignancies. However, T cells and NK cells often exhibit poor infiltration and are readily suppressed by the immunosuppressive TME in solid tumors [ 24 , 151 ]. In contrast, MSCs naturally home to inflammatory and injured tissues and demonstrate superior tumor penetration. These core attributes render CAR-MSCs particularly well-suited for the treatment of autoimmune diseases and the stromal microenvironment of solid tumors. Current evidence suggests that CAR-MSCs hold the potential to become a transformative platform in the field of cell therapy.

Preclinical studies on CAR-MSCs

In GvHD

Recently, researchers at the Mayo Clinic have developed an innovative immunotherapy strategy that may lay the foundation for treating various immune diseases [ 12 , 103 ]. This novel approach involves integrating CARs with MSCs, leveraging synthetic biology to engineer MSCs into specialized cell types known as CAR-MSCs. The development of CAR-MSCs enhances their ability to home to target tissues, which is mediated by a tissue-specific antigen-binding domain and further supported by the immunosuppressive properties conferred through the incorporation of an immunosuppressive intracellular signaling domain. The bioengineering process of CAR-MSCs has been validated as both safe and effective in maintaining stem cell characteristics, achieving a transduction efficiency exceeding 80% for the CAR construct in human AD-MSCs.

Researchers have utilized CD28ζ technology to genetically modify CAR-MSCs, enabling them to target E-cadherin (Ecad) for the treatment of GvHD. Ecad serves as a critical target for activated T cells in immune-related diseases. The modified CAR-MSCs demonstrated specific immunosuppressive effects in both in vitro and in vivo experiments, outperforming traditional MSC treatments. CAR-MSC intervention significantly alleviated symptoms in mice, suppressed T-cell activity, and increased survival rates. These findings indicate that the design of specific CAR constructs can generate CAR-MSCs with enhanced immunosuppressive capabilities while maintaining nontoxicity.

In tumor intervention

CAR-MSCs combine the targeting specificity of CARs with the immunomodulatory and tissue repair capabilities of MSCs, aiming to enhance therapeutic efficacy while minimizing side effects. This innovative strategy offers promising new avenues for the treatment of malignant tumors, particularly solid tumors. Currently, several studies have explored the application of CAR-MSCs in various tumor models, including GBM, Ewing’s sarcoma (ES), AML, and breast cancer, yielding encouraging results.

Golinelli et al. developed bifunctional MSCs that simultaneously express TRAIL and anti-GD2 CAR (GD2 tCAR), which can selectively target and eliminate GD2-positive GBM cells [ 13 ]. TRAIL is a ligand capable of inducing apoptosis in malignant cells but is relatively safe for normal cells [ 152 ]. Tumors of neuroectodermal origin, such as GBM, sarcoma, and neuroblastoma, frequently overexpress GD2 [ 153 ]. In the ES model, bifunctional MSCs can effectively target and eliminate ES cells, particularly demonstrating significant efficacy in the lung metastasis model [ 14 ]. In the treatment of leukemia, Aliperta et al. modified the single-cell-picked clone 1 (SCP-1) MSC line to continuously secrete CD33/CD3 bispecific antibodies (bsAbs), thereby redirecting T-cell toxicity to CD33 + leukemia cells and enhancing tumor-specific killing effects [ 154 ]. The findings of these studies collectively demonstrate that the MSC-based platform can function as a modular biomanufacturing system for precision immunotherapy. Unlike the aforementioned research preparation techniques, Tang S et al. employed a non-viral method to engineer iPSCs with stable overexpression of NKG2D-CAR, which were subsequently differentiated into MSCs. In a murine model of lung cancer, these NKG2D-CAR-expressing iPSC-derived MSCs exhibited enhanced tumor tropism and robust proliferative capacity [ 155 ]. Despite the limited number of studies on CAR-MSC treatment, their significant potential in breast cancer therapy has been highlighted in the literature. CAR-MSCs represent a promising therapeutic option for breast cancer and merit further exploration [ 156 ].

The findings from these studies suggest that genetically engineered MSCs expressing CARs exhibit robust anticancer and immunosuppressive properties (Table 3 ), mediated by targeting precision and immunomodulatory capabilities via gene transfer mechanisms and cytokine secretion (Fig. 3 ).

Table 3 CAR-MSCs in preclinical trials MSC Origin CAR Structure Disease Optimization Ref. Human adipose tissue EcCAR containing CD28ζ GvHD Immunosuppression; tissue regeneration; trafficking [ 103 ] Human adipose tissue truncated anti-GD2 CAR with TRAIL GBM Targeting potential; anticancer activity [ 13 ] Human adipose tissue truncated anti-GD2 CAR with sTRAIL ES Tumor targeting; persistence [ 14 ] SCP-1 cell line from human bone marrow anti-CD33-anti-CD3 bsAb with CD137L AML T-cell redirection; facilitation of T-cell-mediated elimination of autologous leukemic cells; immune response [ 154 ] iPSCs NKG2D-CAR NSCLC Migration and adhesion; tumor-homing ability [ 155 ] MSCs, mesenchymal stem cells; EcCAR, E-cadherin chimeric antigen receptor; GvHD, graft-versus-host disease; sTRAIL, soluble TNF-related apoptosis-inducing ligand; GBM, glioblastoma; ES, Ewing sarcoma; SCP-1, single-cell-picked clone 1; AML, acute myeloid leukemia; iPSCs, induced pluripotent stem cells; NSCLC, non-small cell lung cancer; Ref., reference

Fig. 3 Schematic of CAR-MSC architecture and potential mechanisms of action in disease contexts. MSCs express CARs via gene transduction, thereby enhancing their tissue-targeting and immunoregulatory capabilities. Different ScFvs in the extracellular domain of CARs enable the targeting of distinct tissues, while varying intracellular signaling domains can result in differential functional enhancements, including immunomodulation, sustained proliferation, enhanced secretion, or cytotoxicity. Abbreviations: MSCs, mesenchymal stem cells; CAR, chimeric antigen receptor

Clinical studies on CAR-MSCs

Despite the promising therapeutic potential of CAR-MSCs, research on their antitumor efficacy and immunomodulatory properties remains in its nascent stages with a limited scope. Leveraging their promising results, Mayo Clinic’s Cellular Engineering Laboratory—the pioneer in CAR-MSC development—is designing a phase I trial to evaluate Ecad-CAR-MSCs with CD28ζ signaling in the treatment of steroid-refractory acute GvHD, with plans for subsequent multicenter studies in collaboration with industry partners [ 103 ]. However, detailed information regarding the survey (including inclusion/exclusion criteria, primary and secondary endpoints, and study duration) currently lacks supporting data.

To explore the use of MSCs in the treatment of GvHD, a series of randomized clinical trials led by our center (ChiCTR-IOR-15006330; ChiCTR-IIR-16007806; ChiCTR1900022292) has been conducted over a decade to produce three major achievements. In the first trial, we established that repeated UC-MSC infusions initiated at the 100-day mark post-transplantation markedly lowered the incidence and severity of cGvHD (27.4% vs. 49.0%; P = 0.02) [ 74 ]. The subsequent investigation demonstrated protective effects with early-phase infusion repetition from day + 45 + 81 posttransplant, resulting in a remarkable reduction in the cGvHD rate (5.4% vs. 17.4%; P = 0.03) [ 95 ]. Most recently, the third clinical trial extended this paradigm by evaluating sequential MSC infusions initiated within the first 3 months post-HSCT, representing the earliest intervention window among these studies [ 10 ]. Collectively, these phased clinical investigations substantiate that prophylactic MSC administration effectively reduces the incidence of cGVHD, with an excellent safety profile. CARs can be applied in future work on the basis of previous clinical trials by leveraging their antigen specificity and immune activation. Specifically, we aim to explore and compare the effectiveness of CAR-modified MSCs in further reducing GvHD at different time points.

CAR-MSCs may serve as an affordable “off-the-shelf” cell therapy platform applicable for the treatment of not only GvHD but also other immune-mediated diseases, such as Crohn’s disease and ulcerative colitis. As of August 2025, no clinical studies on CAR-MSC therapy have been registered on international clinical trial platforms, including ClinicalTrials.gov, WHO ICTRP, EudraCT, and ChiCTR. This gap highlights the urgent need for more clinical research to facilitate the translational application of CAR-MSCs.

Advances in CAR-MSC manufacturing technologies

The technology used to manufacture CAR-MSCs is critical for their clinical application and involves five main aspects: [ 1 ] optimization of the CAR structure; [ 2 ] stringent standards for MSC isolation; [ 3 ] precise gene editing; [ 4 ] GMP-compliant amplification; and [ 5 ] comprehensive quality assessment (including efficacy, identity testing, and sterility). While the production of CAR-MSCs can leverage existing technologies for CAR-T-cell therapy, parameter optimization must still be tailored to their unique characteristics.

First, compared with that of CAR-T cells, the design of the CAR structure for CAR-MSCs must consider the unique biological characteristics of MSCs. For example, MSCs exhibit low immunogenicity and broad regulatory functions; however, their antigen-specific recognition and cytotoxic capabilities are relatively limited. Consequently, molecular modules that increase migration and homing abilities are frequently incorporated into CAR designs.

Second, MSCs can be isolated from various tissues. However, owing to the limitations associated with BM collection, AD- or UC-MSCs may become more prevalent in the future. Collected samples are subjected to isolation and adherent culture in specialized MSC expansion medium under controlled conditions (5% CO 2 and 37 °C) to obtain a high-purity cell population.

Third, the core of CAR-MSC manufacturing involves introducing the CAR gene into MSCs. Common gene modification techniques include viral vector-mediated transduction and nonviral vector-mediated transfection. Viral vectors, such as lentiviruses, are widely used because of their high integration efficiency and stable expression. Lentiviral vectors have been successfully applied multiple times in CAR-MSC preparations [ 13 , 14 , 103 , 154 ], enabling efficient CAR gene transduction while preserving MSC proliferation and differentiation capabilities. However, it is crucial to consider the impact of the viral titer and cell condition on transduction efficiency, as well as the potential carcinogenic risks and high costs associated with this approach. Researchers are developing new delivery systems, such as adeno-associated virus (AAV) and integrase-deficient lentiviral vectors (IDLVs), to improve viral vector design. These systems provide greater transduction efficiency, a reduced immune response, lower production costs, and safe genomic integration [ 157 , 158 ]. Recently, Ye et al. developed an innovative gene delivery platform that combines Sleeping Beauty (SB) transposase mRNA with AAV-encoded SB transposons containing therapeutic transgenes [ 159 ]. The platform enables efficient CAR delivery to T cells while also achieving the successful transduction of NK cells, myeloid lineage cells, and iPSCs. Comparative analyses have revealed advantages over lentiviral vectors and electroporation-based delivery, including sustained transgene expression, enhanced activity, increased cell yields, and increased viability. This system is designed to address current limitations in cell engineering, particularly for MSC therapies that require persistent CAR expression. Although nonviral vectors (such as electroporation, nanoparticle, and transposon systems) have not been extensively reported for CAR-MSC preparation, they offer advantages such as low toxicity, high biocompatibility, and ease of scale-up production, warranting further exploration. Gene editing technologies, such as CRISPR, offer expanded opportunities for enhancing immunomodulatory functions or improving specific antigen recognition through the precise modification of the MSC genome [ 158 , 160 ].

Fourth, genetically modified MSCs need to be expanded in vitro to obtain a sufficient quantity. Although MSCs have a strong ability to expand, multiple passages may lead to phenotypic and functional changes. Studies suggest that MSCs can be cultured for up to 20 passages [ 103 ], with genetic modifications using MSCs between P8 and P13 being viable [ 14 ]. As reported, human MSCs have been immortalized through lentiviral transduction via the gene encoding human telomerase reverse transcriptase (hTERT) [ 154 ]. In addition, cell proliferation and functional integrity can be maintained by optimizing culture conditions via the addition of growth factors or cytokines.

Fifth, stringent quality control protocols are indispensable in CAR-MSC manufacturing, encompassing comprehensive assessments of CAR surface expression uniformity (via flow cytometry or immunofluorescence), cellular purity, functional potency (proliferation kinetics, apoptosis resistance, targeted migration efficiency), and microbiological safety (sterility testing, mycoplasma screening). The implementation of these analytical benchmarks guarantees that therapeutic-grade cell products meet clinical translation requirements.

Despite notable advancements in CAR-MSC manufacturing technology, critical challenges persist in achieving clinical-grade cell products. Overcoming technical barriers, including suboptimal gene delivery efficacy, off-target risks, process variability across GMP facilities, and the implementation of phase-appropriate quality control platforms, remains imperative for successful translation from preclinical development to investigational new drug (IND)-enabling studies.

Potential challenges and strategies for CAR-MSC applications

Despite the enthusiasm elicited by preclinical studies reporting the potential of CAR-MSCs, the clinical translation of CAR-MSC applications has proven to be more challenging than expected. Owing to the limitations of cell therapies such as CAR-T cells and CAR-NK cells [ 161 ], several factors might pose potential difficulties for future researchers to focus on for improving the development of CAR-MSCs (Fig. 4 ). These include the following: [ 1 ] Ensuring the targeting ability of CAR-MSCs to the greatest extent possible. This might involve chemokine receptor expression, genetic manipulation of MSCs, and modification of MSC membranes. Additionally, the variance in the CAR structure is also a significant factor influencing the targeting force [ 2 ]. Further augmenting the antitumor and immunosuppressive effects of CAR-MSCs [ 3 ]. Prolonging the durability of CAR-MSCs. Although MSCs are highly desirable for reducing inflammation and promoting tissue regeneration, the harsh microenvironment in disease states can compromise cell survival, and the limited amplification capacity and sustainability of MSCs restrict their broader application [ 4 ]. Comprehensively optimizing the process of CAR-MSC preparation to attain the maximum therapeutic effect from the perspective of MSC source selection, culture protocol, condition, cryopreservation, and thawing; diverse donor characteristics, such as age and sex; type of CAR vector, e.g., viral or nonviral; and other factors [ 5 ]. Selecting the method of CAR-MSC administration for different diseases, as well as standardizing the treatment dose and frequency. In addition, the future clinical application of CAR-MSCs also calls for strict regulatory and approval procedures to ensure their safety and efficacy.

Fig. 4 Potential challenges and strategic optimization for the clinical translation of CAR-MSCs. This multilayer pie chart provides a comprehensive overview of the potential challenges and anticipated strategies for addressing these challenges in the future development of CAR-MSCs. The middle layer highlights five major obstacles to CAR-MSC development and clinical translation, each denoted by a distinct color. The outermost layer, extending from each of the five segments, details specific solutions corresponding to these challenges. Abbreviations: MSCs, mesenchymal stem cells; CAR, chimeric antigen receptor; PCPs, positively charged patches

Increasing the targeted efficacy of CAR-MSCs

MSCs are valued for their tumor tropism. Studies have shown that MSCs respond to the selective recruitment of new cells induced by the TME during different stages of tumor growth, thereby promoting their migration to the tumor site after intravenous injection. This tumor tropism may involve three mechanisms, representing the main strategies for enhancing MSC tumor tropism identified in recent research. First, tumor cells secrete various chemokines and cytokines, such as monocyte chemotactic protein-1 (MCP-1), platelet-derived growth factor (PDGF), and VEGFα, into the TME. MSCs express receptors for these factors on their cell membranes, enabling them to be recruited to the tumor site [ 162 , 163 ]. Second, the metabolic characteristics of the TME serve as a powerful driving force for MSC migration to the tumor site. Hypoxia can upregulate MCP-1 expression by increasing nitric oxide and HIF-1α production, which enhances MSC tumor tropism [ 164 ]. Third, MSCs express adhesion molecules on their membranes, such as vascular cell adhesion molecule-1, intercellular cell adhesion molecule-1, and activated leukocyte cell adhesion molecules, which are also expressed by leukocytes, monocytes, and DCs. These findings indicate that MSCs might share similar migration mechanisms with these immune cells [ 164 ].

In addition, studies have suggested that MSCs expanded in vitro for prolonged periods exhibit a weaker tumor-homing ability than freshly isolated MSCs do, with lower expression levels of CXCR4, possibly partially attributed to the influence of in vitro expansion conditions [ 165 , 166 ]. Therefore, the migration and homing abilities of MSCs can be enhanced by pretreatment with growth factors (such as insulin-like growth factor 1 (IGF-1) and HGF) or cytokines (such as IL-3 and IL-6) to upregulate CXCR4 expression [ 167 , 168 ]. Inserting the CXCR4-encoding gene via viral vectors is an alternative approach [ 169 ]. In summary, optimizing the tumor tropism of MSCs requires consideration of not only their intrinsic characteristics (internal factors) but also tumor features (external factors). The former encompasses factors such as heterogeneity within the cell population and molecular expression patterns involved in migration, whereas the latter includes aspects such as tumor type, tumor surface marker patterns, tumor burden, oxygenation status, and inflammatory state [ 170 , 171 ].

Increasing the targeting ability of CAR-MSCs via CAR modification is crucial for obtaining functional CAR-MSCs. On the basis of strategies for optimizing CAR-T-cell targeting efficiency, the following potential approaches can be considered [ 1 ]. Dual (multiple) specific CARs: Designing CAR molecules to target two or more antigens simultaneously can mitigate antigen escape [ 172 ]. For example, the tandem CAR-T (tanCAR-T) strategy enhances targeting efficacy by linking recognition domains for antigens such as HER2 and IL13Rα2 [ 173 ] [ 2 ]. Boolean logic gates: Using AND, NOT, and OR logic gates improve CAR specificity [ 174 , 175 ]. For example, the AND gate requires the coexistence of two distinct antigens to activate CAR-T cells, reducing off-target risk [ 176 ]. The synNotch receptor recognizes tumor-associated factors and induces the expression of a CAR that can recognize a second TAA, activating carrier cells [ 177 ] [ 3 ]. Exploiting the TME: Engineering CAR-MSCs to express chemokine receptors upregulated in target cells enhances MSC infiltration [ 178 , 179 ] [ 4 ]. Masking CARs: Adding a masking peptide and protease-sensitive linker conceals the antigen-recognition domain until protease-mediated hydrolysis exposes it, leveraging high protease activity in the TME for improved targeting accuracy [ 180 ]. In addition, Volta et al. designed adaptor CAR-T cells (AdFITC-CAR) that specifically target fluorescein-labeled diabody adaptors binding to AML antigens. This system enhances targeting specificity and therapeutic efficacy against AML, demonstrating potential for antigen-versatile CAR-T-cell therapies [ 181 ]. Through continuous development of the CAR design, the improvement of signal transduction mechanisms, and the performance of rigorous clinical trials and validations, the targeting ability of CARs is expected to be significantly enhanced, offering a more promising and feasible outlook for the application of CAR-MSC therapy in the treatment of diseases.

Enhancing the anticancer or immunosuppressive efficacy of CAR-MSCs

While MSCs possess potential for anticancer therapy, their effects on tumors are dualistic. They can inhibit tumor growth through immune modulation and disruption of the TME. They might also facilitate tumor progression by potentially transforming into cancer-initiating cells and fostering a microenvironment that promotes proliferation and metastasis [ 9 ]. This underscores the complexity and challenges of using MSCs in cancer therapy. Future studies should therefore aim to clarify MSC mechanisms in cancer to establish safe and effective therapeutic strategies.

The following section outlines strategies to enhance the anticancer and immunomodulatory effects of CAR-MSCs [ 1 ]. Integration of domains or costimulatory molecules: The immunosuppressive capability of CAR-MSCs can be enhanced by incorporating the functional CD28ζ domain (which has been shown to outperform CD28 and CD3ζ alone) into the CAR [ 103 ]. This incorporation allows CAR-MSCs to transmit signals more efficiently and trigger an immune response upon recognition of target cells. Furthermore, in the design of CAR-MSCs, incorporating costimulatory molecules such as 4−1BBL can significantly enhance their cytotoxic effects on AML cells [ 154 ]. This strategy is linked to T-cell activation, thereby enhancing anticancer effects while minimizing damage to normal tissues. Clearly, the design of CARs and the optimization of corresponding CAR-MSCs warrant further investigation to achieve maximal efficacy across various diseases. Additionally, it remains unclear whether CAR-MSCs can interact with other immune cells to augment therapeutic outcomes [ 2 ]. Gene transmission and expression: Previous studies have shown that MSCs can be engineered to express TRAIL, thereby increasing their antitumor capacity [ 13 , 14 ]. This has prompted researchers to consider CAR-MSCs as carriers for gene delivery, enabling the continuous or direct transfer of therapeutic genes into target cells to optimize treatment efficacy. Leveraging naturally occurring mutations represents a promising strategy. Garcia J et al. identified a gene fusion, CARD11-PIK3R3, in cutaneous T-cell lymphoma that markedly enhances the activity of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activating protein-1 (AP-1) signaling pathways, increases IL-2 production, and contributes to counteracting T-cell dysfunction. This fusion preserves phenotype memory and enhances antitumor efficacy [ 182 ]. This discovery offers a novel perspective on how insights gained from the evolution of malignant cells can improve a wide range of cell therapies [ 3 ]. MSCs can secrete exosomes that contain immunoregulatory molecules capable of modulating the behavior of immune cells [ 183 – 185 ]. CAR-MSCs can harness this mechanism to deliver specific immunoregulatory factors, thereby suppressing immune responses or enhancing antitumor immunity [ 4 ]. MSCs have the inherent ability to home to damaged tissues or tumors, allowing them to migrate directly to the TME [ 186 ]. This property can be harnessed to increase the accumulation of CAR-MSCs at the tumor site, thereby enhancing their anticancer efficacy [ 5 ]. As previously mentioned, by incorporating CAR technology, CAR-MSCs can recognize and bind to specific markers on the surface of diseased cells, such as E-cad, which serves as a target for overactive T cells in immune-related diseases [ 103 ]. This targeting capability enables CAR-MSCs to precisely identify and locate tumor cells or diseased cells.

There is undoubtedly significant potential for CAR-MSCs in both treating cancer efficacy and regulating the immune system. However, research in this field remains in progress, and further studies are needed to investigate the roles of differently CAR-MSC designs in various cancer types and diseases.

Reinforcing the proliferative capacity and persistence of CAR-MSCs

Proliferation refers to the ability of CAR-MSCs to divide in vitro or in vivo and is influenced by culture conditions, gene editing efficiency, MSC source, and cell state. Persistence refers to the duration for which CAR-MSCs survive and function in vivo. Proliferation and persistence are correlated; strong proliferation can enhance in vivo impact and persistence. However, persistence also depends on the microenvironment, antigen stimulation, and immune rejection.

Tonic signaling—the spontaneous activation of chimeric antigen receptors (CARs) in the absence of tumor antigen stimulation—plays a critical role in determining CAR-T-cell therapeutic efficacy. Chen et al. identified that positively charged patches (PCPs) on the CAR antigen-binding domain promote receptor clustering and initiate tonic signaling [ 187 ]. For CAR constructs exhibiting high basal signaling activity (such as GD2-CAR or CSPG4-CAR), strategies like reducing PCP numbers or elevating ionic strength during in vitro expansion can effectively suppress spontaneous activation and mitigate T-cell exhaustion. Conversely, adding PCPs to CARs with low tonic signaling (e.g., CD19 CAR) enhances in vivo persistence. These findings offer a strategy for improving CAR-MSC durability and adaptability through PCP modulation. Preventing CAR protein degradation and promoting CAR recycling may increase CAR-MSC persistence. Li W et al. reported that tumor antigen engagement triggers rapid CAR ubiquitination, leading to CAR downmodulation [ 188 ]. By mutating the lysine in the CAR cytoplasmic domain (CAR KR ) to inhibit ubiquitination, lysosomal degradation of the CAR protein is prevented. These findings suggest that CAR KR -MSCs can reduce surface CAR loss, increasing long-term viability. The screening and identification of mature, safe candidate drugs that can be combined with CAR-MSCs to increase persistence and cytotoxicity represent promising strategies. A recent study demonstrated the antitumor efficacy of afatinib (AFA)-pretreated CAR-T cells in murine leukemia models [ 189 ]. Coculturing AFA with CAR-T cells during preparation revealed that AFA inhibits the TCR and PI3K/AKT/mTOR signaling pathways, reducing cell exhaustion and enhancing persistence and cytotoxicity. These findings provide valuable insights for future research on the development of pretreatment drugs to improve CAR-MSC antitumor cytotoxicity.

Unlike previous studies on CAR-T-cell regulation, Liu Y et al. combined a genetic code expansion system with CAR-T-cell therapy by introducing a stop codon into the CAR protein. The CAR protein expression and cytotoxicity of CAR-T/CAR-NK cells are controlled via an exogenous unnatural amino acid (BOCK). Specifically, BOCK activates CAR-mediated cytotoxicity; without BOCK, CAR activity is inhibited, keeping cells quiescent. Notably, BOCK deficiency reduces tonic signaling and CAR-T-cell exhaustion during in vitro preparation, increasing CAR-T-cell persistence and efficacy [ 190 ]. This reversible BOCK-modified CAR system offers an innovative approach for CAR-MSC development. This strategy improves both proliferation control and durability, optimizing treatment outcomes.

Researchers have explored the expansion of CAR-T cells via the use of engineered cytokines such as IL-15 and IL-7 and the transcription factor Forkhead box protein O1 (FOXO1) [ 191 ]. A recent study revealed that coexpressing IL-15 in GPC3-targeted CAR-T cells enhances their expansion, intratumoral survival, and antitumor activity in solid tumor patients [ 192 ]. Two recent studies identified FOXO1 as a factor that improves CAR-T-cell fitness and efficacy against solid tumors. Doan AE et al. demonstrated that FOXO1 overexpression induces a T-cell memory-associated gene expression program, conferring memory potential and metabolic fitness to CAR-T cells and enhancing in vivo persistence and tumor control [ 193 ]. Chan JD et al. revealed that FOXO1 overexpression promotes a stem-like phenotype in CAR-T cells, which is correlated with improved mitochondrial fitness, persistence, and therapeutic efficacy in vivo [ 194 ]. These findings suggest that genetically enhancing FOXO1 expression in CAR-MSCs is a promising strategy to improve proliferation, durability, and efficacy via favorable metabolic changes and high transformation potential. Furthermore, key transcription factors, such as TCF1, BACH2, MYB, and TOX, are critical in defining and maintaining the exhausted T-cell phenotype. Recent CAR-T studies have highlighted their importance in molecular interactions [ 195 ]. Specifically, bromodomain protein 4 (BRD4) inhibitors alleviate CAR-T-cell exhaustion and inhibit the terminal differentiation of exhausted T cells by downregulating BATF and upregulating EGR1 expression [ 196 ]. A deeper understanding of the molecular mechanisms underlying exhausted T cells could reveal innovative therapeutic strategies for CAR-MSCs.

Lee SS et al. recently demonstrated that MSCs briefly exposed to tropoelastin display an upregulation of proliferation-related genes and a concurrent downregulation of senescence-related genes. Importantly, the senoprotective benefits of tropoelastin persist throughout continuous and long-term MSC culture and significantly extend the replicative lifespan of MSCs [ 197 ]. The use of genetic engineering to modify the genome of MSCs is also a potent approach to increase the proliferation ability of CAR-MSCs or endow them with antiapoptotic characteristics. Jin HJ et al. reported that hUCB-MSCs with high expression of CD146 presented a greater growth rate and greater multidirectional differentiation, expression of stem cell markers, and telomerase activity [ 198 ], indicating that upregulation of the level of CD146 could contribute to promoting the proliferative capacity of CAR-MSCs. In addition, the overexpression of MIF rejuvenated aged MSCs through the activation of autophagy. The abrogation of miR-195 can activate telomere relengthening by increasing the expression levels of telomerase reverse transcriptase (Tert), thereby contributing to the rejuvenation of MSC senescence [ 199 ]. The overexpression of Erb-B2 receptor tyrosine kinase 4 (ERBB4) in senescent MSCs can reverse their aging phenotype and strengthen their resistance to oxidative stress [ 200 ]. The overexpression of Yes-associated protein (YAP) or forkhead box D1 (FOXD1) is also capable of rejuvenating hMSCs [ 201 ].

Strengthening the microenvironmental adaptability of CAR-MSCs enhances their durability. For example, low-oxygen culture inhibits MSC aging, preserves stem cell characteristics, and improves growth dynamics [ 202 ]. Studies have shown that supplementing media with fibroblast growth factor 2, PDGF-BB, ascorbic acid, and epidermal growth factor significantly increases MSC expansion in vitro [ 203 ]. Recent findings confirmed that the overexpression of sirtuins (SIRT1, SIRT3, SIRT6, and SIRT7) delays MSC functional decline and increases MSC differentiation capacity [ 204 – 207 ]. Improving the immunosuppressive environment, providing continuous antigenic stimulation, and combining therapies (e.g., anti-PD-1 blockade) may increase CAR-MSC persistence [ 151 ]. In conclusion, enhancing the proliferation capacity and durability of CAR-MSCs is a complex and challenging task that demands comprehensive and meticulous consideration from multiple perspectives.

Optimization of CAR-MSC production

Selection of MSC source

In CAR-MSC production, selection of the MSC source is a critical factor, with common sources including BM, UC tissue, adipose tissue, placental tissues, and others (Table 4 ). BM is one of the most frequently used MSC sources and is typically harvested from regions such as the iliac crest and sternum. However, BM-MSCs exhibit a limited proliferative capacity that decreases markedly with age, reducing both cell quantity and proliferative potential. The UC, an accessory tissue present at birth, is rich in MSCs. In terms of cellular abundance, the UC contains the highest concentration of MSCs among tissues. Moreover, UC-MSCs demonstrate a short doubling time, a strong colony-forming ability, and a multilineage differentiation potential that is retained even after 40 passages. These cells exhibit rapid proliferation and potent immunomodulatory capabilities. Adipose tissue, which is rich in both adipocytes and MSCs, can be harvested with minimal donor discomfort. AD-MSCs display robust in vitro expansion capabilities and low immunogenicity, making them highly suitable for CAR-MSC production. The amniotic membrane and placenta serve as alternative sources of MSCs, which exhibit superior immunomodulatory properties compared with those of BM- and AD-MSCs [ 208 ].

Table 4 Characteristics of MSCs derived from diverse sources Tissue Source Proliferation Differentiation Immunomodulation Availability and Ethicality BM-MSCs + Advantages of three-line and vascular cell differentiation Relatively weak immunomodulatory effects compared with AD-MSCs Invasive, risk of patient discomfort, low cell content; ethically controversial AD-MSCs ++ Adipogenic differentiation Advantages in regulating DCs Invasive, risk of patient discomfort, high cell content; ethically controversial UC-MSCs +++ More efficient osteogenic differentiation Stronger immunomodulatory effects and lower immunogenicity Noninvasive, painless, higher cell content; less ethically controversial PL-MSCs ++ Better performance in adipogenic and vascular cell differentiation Relatively weak inhibitory effect on T-cell proliferation Noninvasive, painless, rich cell content; less ethically BM-MSCs: bone marrow mesenchymal stem cells; AD-MSCs: adipose-derived mesenchymal stem cells; UC-MSCs: umbilical cord-derived mesenchymal stem cells; PL-MSCs: placenta-derived mesenchymal stem cells; DCs: dendritic cells; Ref., reference

Accessibility and safety are critical considerations that should not be ignored, as these properties vary among MSCs derived from different tissue sources. Obtaining BM-MSCs requires BM aspiration, which may cause harm. In contrast, AD- and UC-MSCs are easier to acquire with minimal donor damage. Certain sources, such as embryonic tissue, may raise ethical and legal concerns during selection. The phenotypic, morphological, and functional characteristics of MSCs can be modulated by various donor-specific variables, including age, gender, body mass index, and overall health condition [ 209 ]. Fetal MSCs exhibit superior properties compared to adult MSCs, including lower immunogenicity, stronger anti-inflammatory and proliferative properties, colony formation ability, osteogenic differentiation ability, higher osteogenic gene expression, and longer telomeres [ 210 ]. In brief, for CAR-MSC production, optimal sources should be evaluated on the basis of cellular traits, procurement feasibility and safety, and disease-specific needs. Stringent quality controls are essential to ensure consistency and therapeutic potential.

Freezing‒thawing of MSCs

MSCs are highly vulnerable to cryopreservation-induced apoptosis. During the freezing‒thawing process, a significant proportion of MSCs lose metabolic activity, and their immunosuppressive function and adhesion capacity are markedly impaired, posing a major technical challenge for obtaining sufficient active cells for therapeutic use [ 211 , 212 ]. Thus, modulating biochemical pathways involved in postthaw degradation, particularly apoptosis, could increase the safety and efficacy of CAR-MSC applications, facilitating their development into economically viable and sustainable therapeutic options. Several studies have shown that Rho-associated kinase (ROCK) inhibitors improve the viability and physiological functions of cryopreserved stem cells and T cells after thawing [ 213 , 214 ], suggesting valuable strategies for optimizing CAR-MSC engineering.

Several strategies have been proposed to reduce defects from the freeze‒thaw process in MSCs [ 215 , 216 ]. These include using rate-controlled freezing devices, optimizing cryoprotectant formulations, and preconditioning MSCs with cytokines (e.g., IFN-γ) before cryopreservation, which enhances their “fast-start” performance after thawing. Cell culture rescue is another approach to improve engineered MSC products. This involves allowing thawed cells to recover under optimal conditions for a defined period to restore metabolic activity. The recovery time typically ranges from 24 to 72 h, depending on the study [ 215 ]. In summary, the optimization of the freeze‒thaw process enables CAR-MSCs to maintain pluripotency, differentiation potential, viability, and cytotoxicity after long-term storage and subsequent in vivo infusion, thereby improving therapeutic efficacy. This advancement is crucial for both cell biology research and clinical applications, providing a solid foundation for future developments.

Nonviral CAR-MSCs May hold promising prospects

In CAR product preparation, the viral vector is a critical raw material. Primary T cells are easy to transduce, but robust CAR transduction in MSCs requires significant optimization. Sirpilla et al. demonstrated that adding protamine sulfate led to high and stable CAR expression in MSCs, resulting in the first successful generation of CAR-MSCs [ 12 ]. The viral vector used for CAR transduction can be mass produced and stored stably at −80 °C for several years [ 217 ]. As final CAR-related products cannot be sterilized by filtration, open processing must be minimized, and quality control tests should ensure the sterility, purity, efficacy, and safety of the vector. Furthermore, selecting viral vectors, monitoring long-term safety, and transitioning to multisite production present challenges for manufacturing CAR-MSCs.

Concerns about the safety, high cost, extended production time, and scalability of viral vectors have led to research into nonviral alternatives for gene delivery [ 218 ]. Compared with traditional viral CAR therapies, nonviral CAR-T therapeutics do not use viral vectors to deliver CAR genes but instead modify T cells through genetic engineering techniques, such as mRNA delivery, programmable nucleases (ZFNs, TALENs, and CRISPR/Cas9) or transposon systems (SB, piggyBac, and Tc Buster), to reduce potential immunogenicity and safety issues [ 219 ]. Moreover, multiple nonviral transfection strategies—such as electroporation, lipid-based nanoparticles, or cell-penetrating peptides—can be employed to deliver CAR-encoding nucleic acids into MSCs, utilizing approaches analogous to those applied in NK cell engineering [ 220 ].

The conclusions of the above studies on nonviral methods for the production of CAR-T/NK cells reflect a highly consistent trend: virus-free CAR immune cell therapies are preferable to traditional viral vector-based therapies. For example, Bexte et al. reported that, compared with those modified by lentiviral vectors, SB-modified NK cells presented high and stable CAR expression and more frequent vector integration into safe genomic harbors. SB-generated CAR-NK cells demonstrated greater cytotoxicity than nontransfected NK cells did [ 221 ]. Additionally, novel hyperactive TcBuster transposase-engineered CAR-NK and CAR-T cells, as reported by Skeate JG et al., presented a low integrated vector copy number, a safe insertion site profile, robust in vitro function, and improved survival in a Burkitt lymphoma xenograft model [ 222 ]. These studies provide valuable insights for the future development of nonviral CAR-MSC engineering to fully realize the therapeutic potential of MSC-based immunotherapy.

Standardization of the CAR-MSC administration protocol

Before CAR-MSCs are used clinically, a standardized protocol must be established for selecting the optimal administration route, determining the dosage, and defining the frequency. This will ensure efficient delivery to target areas and maximize therapeutic potential. The administration route is critical for MSCs to reach tumor tissues. Recent studies have reported methods such as intravenous infusion, intra-arterial injection, intraperitoneal administration, intracranial injection, intranasal delivery, and direct intratumoral injection.

MSCs can be delivered to tumors and metastatic sites via intravenous injection through the vascular system. However, most MSCs (15–30 μm) become trapped in the pulmonary capillary network due to the “first-pass effect” [ 223 , 224 ], limiting delivery efficiency. To overcome this, strategies include the preadministration of vasodilators to widen pulmonary capillaries, although this may reduce the blood supply by lowering blood pressure [ 225 ]. Alternatively, the cultivation of smaller MSCs or the use of MSC-derived exosomes or nanoparticles can enhance delivery [ 226 ]. Injection into specific veins improves organ targeting. For example, Li et al. reported greater homing selectivity with intraparenchymal hepatic vein injection than with intrahepatic or inferior vena cava injection in an acute liver injury model [ 227 ]. Intra-arterial injection of MSCs bypasses the first-pass effect, as most cells reach peripheral tissues before the lungs. This ensures high MSC bioavailability in peripheral organs and makes selective intra-arterial infusion a feasible strategy for targeting specific organs [ 228 ]. However, similar to intravenous administration, the large size of MSCs may cause arterial occlusion, highlighting treatment-related side effects due to their diameter. Owing to the extensive vascular network in the peritoneal cavity, MSCs administered intraperitoneally can access both the lymphatic and circulatory systems [ 229 ]. Kimura et al. reported that, compared with intravenous delivery, intraperitoneal administration of MSCs resulted in more focused localization within neuroblastoma tissue [ 230 ]. This may result from the lack of a first-pass effect in intraperitoneal administration. Furthermore, various routes of administration, such as intracranial (e.g., subarachnoid injection), intranasal, and intracavitary routes, have been explored, each with distinct characteristics. Intracranial administration offers an effective strategy for targeting intracranial tumors with MSCs by bypassing the blood‒brain barrier (BBB). Wang et al. demonstrated in a glioma mouse model that paclitaxel-loaded MSCs delivered via intracranial injection could migrate throughout the entire brain hemisphere [ 231 ]. Intranasal administration offers a noninvasive way to bypass the BBB, showing promise for brain targeting [ 232 ]. Studies have shown that treatment efficacy increases with increasing proximity of the injection site to the tumor. Repeated direct injection of MSCs into a tumor ensures better cell distribution in the TME and improves treatment outcomes [ 233 ].

Systematic regulations for MSC dosing and frequency remain insufficient. The typical infusion dose of MSCs ranges from 1 × 10 6 to 2 × 10 6 cells/kg, varying according to the study design. Infusion regimens may involve single or multiple doses based on the trial objectives. In summary, the administration route, dose, and frequency of CAR-MSCs are determined by the clinical trial design and therapeutic goals. Treatment plans should be tailored to individual patient conditions and study requirements. It is worthwhile to explore the genetic modification of CAR-MSCs, enabling them to express specific genes that increase their proliferation and persistence, potentially leading to a reduction in the required infusion dose in future applications.

Conclusion

The integration of CAR engineering and MSCs represents a novel impetus for cellular immunotherapy. Leveraging advancements in life sciences, the synergistic application of stem cell biology and immunotherapy strategies holds substantial promise for clinical translation. The use of CAR-MSC technology to enhance the endogenous stem cell reservoir and immune cell population may enable precise immune modulation while maintaining long-term systemic homeostasis. However, current CAR-MSC research is still in its infancy. The safety issue involves both the complexity of theoretical deduction and experimental verification, encompassing risks such as tumor formation, long-term survival and genetic stability, non-target tissue damage, and excessive immunosuppression. Breakthroughs in genetic engineering, biomaterial science, and single-cell analysis are expected to accelerate the development of CAR-MSC therapies. Through continued innovation and evidence-based optimization, this emerging therapeutic approach promises to address key challenges in precision medicine and open new avenues for the personalized treatment of malignancies and immune-mediated diseases.

Abbreviations

Associated virus

derived

Afatinib

Protein kinase B

Acute myocardial infarction

Acute myeloid leukemia

Activating protein-1

Acute respiratory distress syndrome

Blood–brain barrier

Bone marrow

Bcl-2-associated X protein

B-cell maturation antigen

Bromodomain protein 4

Bispecific antibodies

Chimeric antigen receptor

Chimeric antigen receptor-engineered mesenchymal stem cells

CUB domain-containing protein 1

Chronic limb-threatening ischemia

Coronavirus disease 2019

Clustered regularly interspaced short palindromic repeats

Cytokine release syndrome

C-X-C motif chemokine ligand 12

C-X-C chemokine receptor type 4

Dendritic cells

Eczema area and severity index

Epidermal growth factor

Estimated glomerular filtration rate

E-cadherin

Ewing sarcoma

Food and drug administration

Fibroblast growth factor

Forkhead box D1

Forkhead box protein O1

Glioblastoma

Good Manufacturing Practice

Graft-versus-host disease

Heart failure with reduced ejection fraction

Hepatocyte growth factor

Hypoxia-inducible transcription factor-1α

Human leukocyte antigen DR

Hematopoietic stem cells

Hematopoietic stem cell transplantation

Human telomerase reverse transcriptase

Intercellular cell adhesion molecule

Integrase-deficient lentiviral vectors

Indoleamine 2,3-dioxygenase

Interferon-γ

Insulin-like growth factor 1

Interleukin

Invariant natural killer T cells

Induced pluripotent stem cells

Proinflammatory macrophage phenotype

Anti-inflammatory macrophage phenotype

Myeloid cell leukemia sequence 1

Monocyte chemotactic protein-1

Major histocompatibility complex

Multiple myeloma

Mammalian target of rapamycin

Mesenchymal stem cells

Nuclear factor kappa-light chain enhancer of activated B cells

Nuclear factor erythroid 2

Neuroglial antigen 2

Natural killer

Neural progenitor cells

Oncolytic adenovirus

Overall survival

Perinatal arterial ischemic stroke

Paired box 5

Positively charged patches

Platelet-derived growth factor

Prostaglandin E2

Phosphoinositide 3-kinase

Reference

Rho-associated kinase

Sleeping beauty

Single-chain variable fragment

Single-cell-picked clone 1

Stromal cell-derived factor-1α

Stanniocalcin-1

Systemic lupus erythematosus

Tumor-associated antigen

T-cell receptor

Transforming growth factor-β

Tumor microenvironment

Tumor necrosis factor

TNF-related apoptosis-inducing ligand

T cells redirected for universal cytokine-mediated killing

Regulatory T cell

Umbilical cord

Vascular cell adhesion molecule

Vascular endothelial growth factor

Yes-associated protein

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Ying Chen, Jing Li and Yingying Ma contributed equally to this work.

Acknowledgements

Not applicable.

Author contributions

C.Z. and X.Z. defined the overarching research direction and provided critical guidance for this study; Y.C. drafted the manuscript; Y.C. and J.J.F. prepared the figures and tables; Y.C., J.L., Y.Y.M., Y.Y., and L.Y. revised the manuscript; all authors have read and approved the final version of the manuscript.

Funding

This study was supported by grants from the National Natural Science Foundation of China (82170212, 82370181, and 82341201), the National Key R&D Program of China (2023YFC2508905), the Chongqing Science and Health Collaborative Major Project on Medical Science and Technology Innovation (2024DBXM004), the Chongqing Medical Scientific Research project (Joint project of Chongqing Health Commission and Science and Technology Bureau) (2025CCXM002), the Clinical Research Special Project of the Second Affiliated Hospital, the Army Medical University (2024F010), and the Special Project for Talent Construction in Xinqiao Hospital of Army Medical University (2022XKRC001).

Data availability

No datasets were generated or analysed during the current study.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

All authors consent to publication.

Competing interests

The authors declare no competing interests.

References

1. Yao P Liu Y-G Huang G Hao L Wang R The development and application of chimeric antigen receptor natural killer (CAR-NK) cells for cancer therapy: current state, challenges and emerging therapeutic advances Exp Hematol Oncol 2024 13 1 118 39633491 10.1186/s40164-024-00583-7 PMC11616395 Yao P, Liu Y-G, Huang G, Hao L, Wang R. The development and application of chimeric antigen receptor natural killer (CAR-NK) cells for cancer therapy: current state, challenges and emerging therapeutic advances. Exp Hematol Oncol. 2024;13(1):118. 39633491 10.1186/s40164-024-00583-7 PMC11616395

2. Zhang C He J Liu L Wang J Wang S Liu L Novel CD19 chimeric antigen receptor T cells manufactured next-day for acute lymphoblastic leukemia Blood Cancer J 2022 12 6 96 35750687 10.1038/s41408-022-00688-4 PMC9232607 Zhang C, He J, Liu L, Wang J, Wang S, Liu L, et al. Novel CD19 chimeric antigen receptor T cells manufactured next-day for acute lymphoblastic leukemia. Blood Cancer J. 2022;12(6):96. 35750687 10.1038/s41408-022-00688-4 PMC9232607

3. Zhang C Wang X-Q Zhang R-L Liu F Wang Y Yan Z-L Donor-derived CD19 CAR-T cell therapy of relapse of CD19-positive B-ALL post allotransplant Leukemia 2021 35 6 1563 70 33077866 10.1038/s41375-020-01056-6 PMC8179843 Zhang C, Wang X-Q, Zhang R-L, Liu F, Wang Y, Yan Z-L, et al. Donor-derived CD19 CAR-T cell therapy of relapse of CD19-positive B-ALL post allotransplant. Leukemia. 2021;35(6):1563–70. 33077866 10.1038/s41375-020-01056-6 PMC8179843

4. Zhou D Zhu X Xiao Y Advances in CAR-T therapy for central nervous system tumors Biomark Res 2024 12 1 132 39506843 10.1186/s40364-024-00679-6 PMC11539471 Zhou D, Zhu X, Xiao Y. Advances in CAR-T therapy for central nervous system tumors. Biomark Res. 2024;12(1):132. 39506843 10.1186/s40364-024-00679-6 PMC11539471

5. Nasiri F Asaadi Y Mirzadeh F Abdolahi S Molaei S Gavgani SP Updates on CAR T cell therapy in multiple myeloma Biomark Res 2024 12 1 102 39261906 10.1186/s40364-024-00634-5 PMC11391811 Nasiri F, Asaadi Y, Mirzadeh F, Abdolahi S, Molaei S, Gavgani SP, et al. Updates on CAR T cell therapy in multiple myeloma. Biomark Res. 2024;12(1):102. 39261906 10.1186/s40364-024-00634-5 PMC11391811

6. Zhang C Liu J Zhong JF Zhang X Engineering CAR-T cells Biomark Res 2017 5 22 28652918 10.1186/s40364-017-0102-y PMC5482931 Zhang C, Liu J, Zhong JF, Zhang X. Engineering CAR-T cells. Biomark Res. 2017;5:22. 28652918 10.1186/s40364-017-0102-y PMC5482931

7. Huang R Li X He Y Zhu W Gao L Liu Y Recent advances in CAR-T cell engineering J Hematol Oncol 2020 13 1 86 32616000 10.1186/s13045-020-00910-5 PMC7333410 Huang R, Li X, He Y, Zhu W, Gao L, Liu Y, et al. Recent advances in CAR-T cell engineering. J Hematol Oncol. 2020;13(1):86. 32616000 10.1186/s13045-020-00910-5 PMC7333410

8. Heo JS Choi Y Kim H-S Kim HO Comparison of molecular profiles of human mesenchymal stem cells derived from bone marrow, umbilical cord blood, placenta and adipose tissue Int J Mol Med 2016 37 1 115 25 26719857 10.3892/ijmm.2015.2413 PMC4687432 Heo JS, Choi Y, Kim H-S, Kim HO. Comparison of molecular profiles of human mesenchymal stem cells derived from bone marrow, umbilical cord blood, placenta and adipose tissue. Int J Mol Med. 2016;37(1):115–25. 26719857 10.3892/ijmm.2015.2413 PMC4687432

9. Zhang Y Wang C Li JJ Revisiting the role of mesenchymal stromal cells in cancer initiation, metastasis and immunosuppression Exp Hematol Oncol 2024 13 1 64 38951845 10.1186/s40164-024-00532-4 PMC11218091 Zhang Y, Wang C, Li JJ. Revisiting the role of mesenchymal stromal cells in cancer initiation, metastasis and immunosuppression. Exp Hematol Oncol. 2024;13(1):64. 38951845 10.1186/s40164-024-00532-4 PMC11218091

10. Yao H Huang R Fu H Lin R Zhang Y Feng Y Sequential infusion of mesenchymal stem cell for Graft-Versus-Host disease prevention in haploidentical hematopoietic stem cell transplantation: an open-label, multicenter, randomized controlled clinical trial J Clin Oncol 2025 43 17 1997 2006 10.1200/JCO-24-02119 40233291 PMC12169861 Yao H, Huang R, Fu H, Lin R, Zhang Y, Feng Y, et al. Sequential infusion of mesenchymal stem cell for Graft-Versus-Host disease prevention in haploidentical hematopoietic stem cell transplantation: an open-label, multicenter, randomized controlled clinical trial. J Clin Oncol. 2025;43(17):1997–2006. 10.1200/JCO-24-02119 40233291 10.1200/JCO-24-02119 PMC12169861

11. Matas J García C Poblete D Vernal R Ortloff A Luque-Campos N A phase I dose-escalation clinical trial to assess the safety and efficacy of umbilical cord-derived mesenchymal stromal cells in knee osteoarthritis Stem Cells Transl Med 2024 13 3 193 203 38366909 10.1093/stcltm/szad088 PMC10940813 Matas J, García C, Poblete D, Vernal R, Ortloff A, Luque-Campos N, et al. A phase I dose-escalation clinical trial to assess the safety and efficacy of umbilical cord-derived mesenchymal stromal cells in knee osteoarthritis. Stem Cells Transl Med. 2024;13(3):193–203. 38366909 10.1093/stcltm/szad088 PMC10940813

12. Sirpilla O Sakemura RL Hefazi M Girsch JH Huynh T Cox MJ Chimeric antigen receptor engineering of mesenchymal stromal cells (CAR-MSC) enhance immunosuppression and outcomes in graft versus host disease (GvHD) preclinical models Blood 2022 140 Supplement 1 1579 80 Sirpilla O, Sakemura RL, Hefazi M, Girsch JH, Huynh T, Cox MJ, et al. Chimeric antigen receptor engineering of mesenchymal stromal cells (CAR-MSC) enhance immunosuppression and outcomes in graft versus host disease (GvHD) preclinical models. Blood. 2022;140(Supplement 1):1579–80.

13. Golinelli G Grisendi G Prapa M Bestagno M Spano C Rossignoli F Targeting GD2-positive glioblastoma by chimeric antigen receptor empowered mesenchymal progenitors Cancer Gene Ther 2020 27 7–8 558 70 30464207 10.1038/s41417-018-0062-x PMC7445885 Golinelli G, Grisendi G, Prapa M, Bestagno M, Spano C, Rossignoli F, et al. Targeting GD2-positive glioblastoma by chimeric antigen receptor empowered mesenchymal progenitors. Cancer Gene Ther. 2020;27(7–8):558–70. 30464207 10.1038/s41417-018-0062-x PMC7445885

14. Golinelli G Grisendi G Dall’Ora M Casari G Spano C Talami R Anti-GD2 CAR MSCs against metastatic ewing’s sarcoma Transl Oncol 2022 15 1 101240 34649148 10.1016/j.tranon.2021.101240 PMC8517927 Golinelli G, Grisendi G, Dall’Ora M, Casari G, Spano C, Talami R, et al. Anti-GD2 CAR MSCs against metastatic ewing’s sarcoma. Transl Oncol. 2022;15(1):101240. 34649148 10.1016/j.tranon.2021.101240 PMC8517927

15. Yan L Li J Zhang C The role of MSCs and CAR-MSCs in cellular immunotherapy Cell Commun Signal 2023 21 1 187 37528472 10.1186/s12964-023-01191-4 PMC10391838 Yan L, Li J, Zhang C. The role of MSCs and CAR-MSCs in cellular immunotherapy. Cell Commun Signal. 2023;21(1):187. 37528472 10.1186/s12964-023-01191-4 PMC10391838

16. Shaw TD Krasnodembskaya AD Schroeder GN Doherty DF Silva JD Tandel SM Human mesenchymal stromal cells inhibit Mycobacterium avium replication in clinically relevant models of lung infection Thorax 2024 79 8 778 87 38508718 10.1136/thorax-2023-220819 PMC11287638 Shaw TD, Krasnodembskaya AD, Schroeder GN, Doherty DF, Silva JD, Tandel SM, et al. Human mesenchymal stromal cells inhibit Mycobacterium avium replication in clinically relevant models of lung infection. Thorax. 2024;79(8):778–87. 38508718 10.1136/thorax-2023-220819 PMC11287638

17. Wang L Deng Z Li Y Wu Y Yao R Cao Y Ameliorative effects of mesenchymal stromal cells on senescence associated phenotypes in naturally aged rats J Transl Med 2024 22 1 722 39103873 10.1186/s12967-024-05486-z PMC11302158 Wang L, Deng Z, Li Y, Wu Y, Yao R, Cao Y, et al. Ameliorative effects of mesenchymal stromal cells on senescence associated phenotypes in naturally aged rats. J Transl Med. 2024;22(1):722. 39103873 10.1186/s12967-024-05486-z PMC11302158

18. Gross G Waks T Eshhar Z Expression of immunoglobulin-T-cell receptor chimeric molecules as functional receptors with antibody-type specificity Proc Natl Acad Sci U S A 1989 86 24 10024 8 2513569 10.1073/pnas.86.24.10024 PMC298636 Gross G, Waks T, Eshhar Z. Expression of immunoglobulin-T-cell receptor chimeric molecules as functional receptors with antibody-type specificity. Proc Natl Acad Sci U S A. 1989;86(24):10024–8. 2513569 10.1073/pnas.86.24.10024 PMC298636

19. Zhang C Wang X Yi H Wang Y Yan Z Zhou J Long-term survival with donor CD19 CAR-T cell treatment for relapsed patients after allogeneic hematopietic stem cell transplantation J Hematol Oncol 2024 17 1 103 39468581 10.1186/s13045-024-01626-6 PMC11520587 Zhang C, Wang X, Yi H, Wang Y, Yan Z, Zhou J, et al. Long-term survival with donor CD19 CAR-T cell treatment for relapsed patients after allogeneic hematopietic stem cell transplantation. J Hematol Oncol. 2024;17(1):103. 39468581 10.1186/s13045-024-01626-6 PMC11520587

20. Tan J Jia Y Zhou M Fu C Tuhin IJ Ye J Chimeric antigen receptors containing the OX40 signalling domain enhance the persistence of T cells even under repeated stimulation with multiple myeloma target cells J Hematol Oncol 2022 15 1 39 35365211 10.1186/s13045-022-01244-0 PMC8974082 Tan J, Jia Y, Zhou M, Fu C, Tuhin IJ, Ye J, et al. Chimeric antigen receptors containing the OX40 signalling domain enhance the persistence of T cells even under repeated stimulation with multiple myeloma target cells. J Hematol Oncol. 2022;15(1):39. 35365211 10.1186/s13045-022-01244-0 PMC8974082

21. Shen C-J Yang Y-X Han EQ Cao N Wang Y-F Wang Y Chimeric antigen receptor containing ICOS signaling domain mediates specific and efficient antitumor effect of T cells against EGFRvIII expressing glioma J Hematol Oncol 2013 6 33 23656794 10.1186/1756-8722-6-33 PMC3658918 Shen C-J, Yang Y-X, Han EQ, Cao N, Wang Y-F, Wang Y, et al. Chimeric antigen receptor containing ICOS signaling domain mediates specific and efficient antitumor effect of T cells against EGFRvIII expressing glioma. J Hematol Oncol. 2013;6:33. 23656794 10.1186/1756-8722-6-33 PMC3658918

22. Han Y Xie W Song D-G Powell DJ Control of triple-negative breast cancer using ex vivo self-enriched, costimulated NKG2D CAR T cells J Hematol Oncol 2018 11 1 92 29980239 10.1186/s13045-018-0635-z PMC6035420 Han Y, Xie W, Song D-G, Powell DJ. Control of triple-negative breast cancer using ex vivo self-enriched, costimulated NKG2D CAR T cells. J Hematol Oncol. 2018;11(1):92. 29980239 10.1186/s13045-018-0635-z PMC6035420

23. Liu J Zhong JF Zhang X Zhang C Allogeneic CD19-CAR-T cell infusion after allogeneic hematopoietic stem cell transplantation in B cell malignancies J Hematol Oncol 2017 10 1 35 28143567 10.1186/s13045-017-0405-3 PMC5282795 Liu J, Zhong JF, Zhang X, Zhang C. Allogeneic CD19-CAR-T cell infusion after allogeneic hematopoietic stem cell transplantation in B cell malignancies. J Hematol Oncol. 2017;10(1):35. 28143567 10.1186/s13045-017-0405-3 PMC5282795

24. Ai K Liu B Chen X Huang C Yang L Zhang W Optimizing CAR-T cell therapy for solid tumors: current challenges and potential strategies J Hematol Oncol 2024 17 1 105 39501358 10.1186/s13045-024-01625-7 PMC11539560 Ai K, Liu B, Chen X, Huang C, Yang L, Zhang W, et al. Optimizing CAR-T cell therapy for solid tumors: current challenges and potential strategies. J Hematol Oncol. 2024;17(1):105. 39501358 10.1186/s13045-024-01625-7 PMC11539560

25. Brocker T Chimeric Fv-zeta or Fv-epsilon receptors are not sufficient to induce activation or cytokine production in peripheral T cells Blood 2000 96 5 1999 2001 10961908 Brocker T. Chimeric Fv-zeta or Fv-epsilon receptors are not sufficient to induce activation or cytokine production in peripheral T cells. Blood. 2000;96(5):1999–2001. 10961908

26. Quintarelli C Orlando D Boffa I Guercio M Polito VA Petretto A Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma Oncoimmunology 2018 7 6 e1433518 29872565 10.1080/2162402X.2018.1433518 PMC5980417 Quintarelli C, Orlando D, Boffa I, Guercio M, Polito VA, Petretto A, et al. Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma. Oncoimmunology. 2018;7(6):e1433518. 29872565 10.1080/2162402X.2018.1433518 PMC5980417

27. Tumino N Weber G Besi F Del Bufalo F Bertaina V Paci P Polymorphonuclear myeloid-derived suppressor cells impair the anti-tumor efficacy of GD2.CAR T-cells in patients with neuroblastoma J Hematol Oncol 2021 14 1 191 34772439 10.1186/s13045-021-01193-0 PMC8588686 Tumino N, Weber G, Besi F, Del Bufalo F, Bertaina V, Paci P, et al. Polymorphonuclear myeloid-derived suppressor cells impair the anti-tumor efficacy of GD2.CAR T-cells in patients with neuroblastoma. J Hematol Oncol. 2021;14(1):191. 34772439 10.1186/s13045-021-01193-0 PMC8588686

28. Guedan S Posey AD Shaw C Wing A Da T Patel PR Enhancing CAR T cell persistence through ICOS and 4-1BB costimulation JCI Insight 2018 3 1 e96976 10.1172/jci.insight.96976 29321369 PMC5821198 Guedan S, Posey AD, Shaw C, Wing A, Da T, Patel PR, et al. Enhancing CAR T cell persistence through ICOS and 4-1BB costimulation. JCI Insight. 2018;3(1):e96976. 10.1172/jci.insight.96976 29321369 10.1172/jci.insight.96976 PMC5821198

29. Till BG Jensen MC Wang J Qian X Gopal AK Maloney DG CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results Blood 2012 119 17 3940 50 22308288 10.1182/blood-2011-10-387969 PMC3350361 Till BG, Jensen MC, Wang J, Qian X, Gopal AK, Maloney DG, et al. CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results. Blood. 2012;119(17):3940–50. 22308288 10.1182/blood-2011-10-387969 PMC3350361

30. Korell F Schönland S Schmitt A Jansen M Farid K Müller-Tidow C First third-generation CAR T cell application targeting CD19 for the treatment of systemic IgM AL amyloidosis with underlying marginal zone lymphoma Biomark Res 2023 11 1 91 37838756 10.1186/s40364-023-00532-2 PMC10576295 Korell F, Schönland S, Schmitt A, Jansen M, Farid K, Müller-Tidow C, et al. First third-generation CAR T cell application targeting CD19 for the treatment of systemic IgM AL amyloidosis with underlying marginal zone lymphoma. Biomark Res. 2023;11(1):91. 37838756 10.1186/s40364-023-00532-2 PMC10576295

31. Zhao Y Chen J Andreatta M Feng B Xie Y-Q Wenes M IL-10-expressing CAR T cells resist dysfunction and mediate durable clearance of solid tumors and metastases Nat Biotechnol 2024 42 11 1693 704 38168996 10.1038/s41587-023-02060-8 Zhao Y, Chen J, Andreatta M, Feng B, Xie Y-Q, Wenes M, et al. IL-10-expressing CAR T cells resist dysfunction and mediate durable clearance of solid tumors and metastases. Nat Biotechnol. 2024;42(11):1693–704. 38168996 10.1038/s41587-023-02060-8

32. Li X Chen T Li X Zhang H Li Y Zhang S Therapeutic targets of armored chimeric antigen receptor T cells navigating the tumor microenvironment Exp Hematol Oncol 2024 13 1 96 39350256 10.1186/s40164-024-00564-w PMC11440706 Li X, Chen T, Li X, Zhang H, Li Y, Zhang S, et al. Therapeutic targets of armored chimeric antigen receptor T cells navigating the tumor microenvironment. Exp Hematol Oncol. 2024;13(1):96. 39350256 10.1186/s40164-024-00564-w PMC11440706

33. Lei W Zhao A Liu H Yang C Wei C Guo S Safety and feasibility of anti-CD19 CAR T cells expressing inducible IL-7 and CCL19 in patients with relapsed or refractory large B-cell lymphoma Cell Discov 2024 10 1 5 38191529 10.1038/s41421-023-00625-0 PMC10774422 Lei W, Zhao A, Liu H, Yang C, Wei C, Guo S, et al. Safety and feasibility of anti-CD19 CAR T cells expressing inducible IL-7 and CCL19 in patients with relapsed or refractory large B-cell lymphoma. Cell Discov. 2024;10(1):5. 38191529 10.1038/s41421-023-00625-0 PMC10774422

34. Liu H Lei W Zhang C Yang C Wei J Guo Q CD19-specific CAR T cells that express a PD-1/CD28 chimeric switch-receptor are effective in patients with PD-L1-positive B-cell lymphoma Clin Cancer Res 2021 27 2 473 84 33028589 10.1158/1078-0432.CCR-20-1457 Liu H, Lei W, Zhang C, Yang C, Wei J, Guo Q, et al. CD19-specific CAR T cells that express a PD-1/CD28 chimeric switch-receptor are effective in patients with PD-L1-positive B-cell lymphoma. Clin Cancer Res. 2021;27(2):473–84. 33028589 10.1158/1078-0432.CCR-20-1457

35. Chmielewski M Kopecky C Hombach AA Abken H IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression Cancer Res 2011 71 17 5697 706 21742772 10.1158/0008-5472.CAN-11-0103 Chmielewski M, Kopecky C, Hombach AA, Abken H. IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression. Cancer Res. 2011;71(17):5697–706. 21742772 10.1158/0008-5472.CAN-11-0103

36. Jin C Ma J Ramachandran M Yu D Essand M CAR t cells expressing a bacterial virulence factor trigger potent bystander antitumour responses in solid cancers Nat Biomed Eng 2022 6 7 830 41 35379957 10.1038/s41551-022-00875-5 PMC9288934 Jin C, Ma J, Ramachandran M, Yu D, Essand M. CAR t cells expressing a bacterial virulence factor trigger potent bystander antitumour responses in solid cancers. Nat Biomed Eng. 2022;6(7):830–41. 35379957 10.1038/s41551-022-00875-5 PMC9288934

37. Gardner TJ Lee JP Bourne CM Wijewarnasuriya D Kinarivala N Kurtz KG Engineering CAR-T cells to activate small-molecule drugs in situ Nat Chem Biol 2022 18 2 216 25 34969970 10.1038/s41589-021-00932-1 PMC9152922 Gardner TJ, Lee JP, Bourne CM, Wijewarnasuriya D, Kinarivala N, Kurtz KG, et al. Engineering CAR-T cells to activate small-molecule drugs in situ. Nat Chem Biol. 2022;18(2):216–25. 34969970 10.1038/s41589-021-00932-1 PMC9152922

38. Rafiq S Yeku OO Jackson HJ Purdon TJ van Leeuwen DG Drakes DJ Targeted delivery of a PD-1-blocking ScFv by CAR-T cells enhances anti-tumor efficacy in vivo Nat Biotechnol 2018 36 9 847 56 30102295 10.1038/nbt.4195 PMC6126939 Rafiq S, Yeku OO, Jackson HJ, Purdon TJ, van Leeuwen DG, Drakes DJ, et al. Targeted delivery of a PD-1-blocking ScFv by CAR-T cells enhances anti-tumor efficacy in vivo . Nat Biotechnol. 2018;36(9):847–56. 30102295 10.1038/nbt.4195 PMC6126939

39. Heitzeneder S Bosse KR Zhu Z Zhelev D Majzner RG Radosevich MT GPC2-CAR T cells tuned for low antigen density mediate potent activity against neuroblastoma without toxicity Cancer Cell 2022 40 1 53 69.e9 10.1016/j.ccell.2021.12.005 34971569 PMC9092726 Heitzeneder S, Bosse KR, Zhu Z, Zhelev D, Majzner RG, Radosevich MT, et al. GPC2-CAR T cells tuned for low antigen density mediate potent activity against neuroblastoma without toxicity. Cancer Cell. 2022;40(1):53–69.e9. 10.1016/j.ccell.2021.12.005 34971569 10.1016/j.ccell.2021.12.005 PMC9092726

40. Qin L Cui Y Yuan T Chen D Zhao R Li S Co-expression of a PD-L1-specific chimeric switch receptor augments the efficacy and persistence of CAR T cells via the CD70-CD27 axis Nat Commun 2022 13 1 6051 36229619 10.1038/s41467-022-33793-w PMC9561169 Qin L, Cui Y, Yuan T, Chen D, Zhao R, Li S, et al. Co-expression of a PD-L1-specific chimeric switch receptor augments the efficacy and persistence of CAR T cells via the CD70-CD27 axis. Nat Commun. 2022;13(1):6051. 36229619 10.1038/s41467-022-33793-w PMC9561169

41. Hirabayashi K Du H Xu Y Shou P Zhou X Fucá G Dual targeting CAR-T cells with optimal costimulation and metabolic fitness enhance antitumor activity and prevent escape in solid tumors Nat Cancer 2021 2 9 904 18 34746799 10.1038/s43018-021-00244-2 PMC8570569 Hirabayashi K, Du H, Xu Y, Shou P, Zhou X, Fucá G, et al. Dual targeting CAR-T cells with optimal costimulation and metabolic fitness enhance antitumor activity and prevent escape in solid tumors. Nat Cancer. 2021;2(9):904–18. 34746799 10.1038/s43018-021-00244-2 PMC8570569

42. Katsarou A Sjöstrand M Naik J Mansilla-Soto J Kefala D Kladis G Combining a CAR and a chimeric costimulatory receptor enhances T cell sensitivity to low antigen density and promotes persistence Sci Transl Med 2021 13 623 eabh1962 34878825 10.1126/scitranslmed.abh1962 PMC9869449 Katsarou A, Sjöstrand M, Naik J, Mansilla-Soto J, Kefala D, Kladis G, et al. Combining a CAR and a chimeric costimulatory receptor enhances T cell sensitivity to low antigen density and promotes persistence. Sci Transl Med. 2021;13(623):eabh1962. 34878825 10.1126/scitranslmed.abh1962 PMC9869449

43. Kagoya Y Tanaka S Guo T Anczurowski M Wang C-H Saso K A novel chimeric antigen receptor containing a JAK-STAT signaling domain mediates superior antitumor effects Nat Med 2018 24 3 352 9 29400710 10.1038/nm.4478 PMC5839992 Kagoya Y, Tanaka S, Guo T, Anczurowski M, Wang C-H, Saso K, et al. A novel chimeric antigen receptor containing a JAK-STAT signaling domain mediates superior antitumor effects. Nat Med. 2018;24(3):352–9. 29400710 10.1038/nm.4478 PMC5839992

44. Yuti P Sawasdee N Natungnuy K Rujirachaivej P Luangwattananun P Sujjitjoon J Enhanced antitumor efficacy, proliferative capacity, and alleviation of T cell exhaustion by fifth-generation chimeric antigen receptor T cells targeting B cell maturation antigen in multiple myeloma Biomed Pharmacother 2023 168 115691 37844355 10.1016/j.biopha.2023.115691 Yuti P, Sawasdee N, Natungnuy K, Rujirachaivej P, Luangwattananun P, Sujjitjoon J, et al. Enhanced antitumor efficacy, proliferative capacity, and alleviation of T cell exhaustion by fifth-generation chimeric antigen receptor T cells targeting B cell maturation antigen in multiple myeloma. Biomed Pharmacother. 2023;168:115691. 37844355 10.1016/j.biopha.2023.115691

45. Zhao W-H Wang B-Y Chen L-J Fu W-J Xu J Liu J Four-year follow-up of LCAR-B38M in relapsed or refractory multiple myeloma: a phase 1, single-arm, open-label, multicenter study in China (LEGEND-2) J Hematol Oncol 2022 15 1 86 35794616 10.1186/s13045-022-01301-8 PMC9261106 Zhao W-H, Wang B-Y, Chen L-J, Fu W-J, Xu J, Liu J, et al. Four-year follow-up of LCAR-B38M in relapsed or refractory multiple myeloma: a phase 1, single-arm, open-label, multicenter study in China (LEGEND-2). J Hematol Oncol. 2022;15(1):86. 35794616 10.1186/s13045-022-01301-8 PMC9261106

46. Yang T Dong Y Zhang M Feng J Fu S Xiao P Prominent efficacy and good safety of sequential CD19 and CD22 CAR-T therapy in relapsed/refractory adult B-cell acute lymphoblastic leukemia Exp Hematol Oncol 2025 14 1 2 39754190 10.1186/s40164-024-00593-5 PMC11697943 Yang T, Dong Y, Zhang M, Feng J, Fu S, Xiao P, et al. Prominent efficacy and good safety of sequential CD19 and CD22 CAR-T therapy in relapsed/refractory adult B-cell acute lymphoblastic leukemia. Exp Hematol Oncol. 2025;14(1):2. 39754190 10.1186/s40164-024-00593-5 PMC11697943

47. Huang H Yu L Weng H Zhang W Wang Z Wang L Advances in CAR-T cell therapy for hematologic and solid malignancies: latest updates from 2024 ESMO congress J Hematol Oncol 2024 17 1 120 39639359 10.1186/s13045-024-01639-1 PMC11622581 Huang H, Yu L, Weng H, Zhang W, Wang Z, Wang L. Advances in CAR-T cell therapy for hematologic and solid malignancies: latest updates from 2024 ESMO congress. J Hematol Oncol. 2024;17(1):120. 39639359 10.1186/s13045-024-01639-1 PMC11622581

48. Yang Y Peng H Wang J Li F New insights into CAR T-cell hematological toxicities: manifestations, mechanisms, and effective management strategies Exp Hematol Oncol 2024 13 1 110 39521987 10.1186/s40164-024-00573-9 PMC11549815 Yang Y, Peng H, Wang J, Li F. New insights into CAR T-cell hematological toxicities: manifestations, mechanisms, and effective management strategies. Exp Hematol Oncol. 2024;13(1):110. 39521987 10.1186/s40164-024-00573-9 PMC11549815

49. Mazinani M Rahbarizadeh F New cell sources for CAR-based immunotherapy Biomark Res 2023 11 1 49 37147740 10.1186/s40364-023-00482-9 PMC10163725 Mazinani M, Rahbarizadeh F. New cell sources for CAR-based immunotherapy. Biomark Res. 2023;11(1):49. 37147740 10.1186/s40364-023-00482-9 PMC10163725

50. Zhang W Huang X In vivo gene editing and in situ generation of chimeric antigen receptor cells for next-generation cancer immunotherapy J Hematol Oncol 2024 17 1 110 39533415 10.1186/s13045-024-01633-7 PMC11559219 Zhang W, Huang X. In vivo gene editing and in situ generation of chimeric antigen receptor cells for next-generation cancer immunotherapy. J Hematol Oncol. 2024;17(1):110. 39533415 10.1186/s13045-024-01633-7 PMC11559219

51. Lu J Ma Y Li Q Xu Y Xue Y Xu S CAR macrophages: a promising novel immunotherapy for solid tumors and beyond Biomark Res 2024 12 1 86 39175095 10.1186/s40364-024-00637-2 PMC11342599 Lu J, Ma Y, Li Q, Xu Y, Xue Y, Xu S. CAR macrophages: a promising novel immunotherapy for solid tumors and beyond. Biomark Res. 2024;12(1):86. 39175095 10.1186/s40364-024-00637-2 PMC11342599

52. Baker DJ Arany Z Baur JA Epstein JA June CH CAR T therapy beyond cancer: the evolution of a living drug Nature 2023 619 7971 707 15 37495877 10.1038/s41586-023-06243-w PMC12522170 Baker DJ, Arany Z, Baur JA, Epstein JA, June CH. CAR T therapy beyond cancer: the evolution of a living drug. Nature. 2023;619(7971):707–15. 37495877 10.1038/s41586-023-06243-w PMC12522170

53. Uccelli A Moretta L Pistoia V Mesenchymal stem cells in health and disease Nat Rev Immunol 2008 8 9 726 36 19172693 10.1038/nri2395 Uccelli A, Moretta L, Pistoia V. Mesenchymal stem cells in health and disease. Nat Rev Immunol. 2008;8(9):726–36. 19172693 10.1038/nri2395

54. Friedenstein AJ Chailakhjan RK Lalykina KS The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells Cell Tissue Kinet 1970 3 4 393 403 5523063 10.1111/j.1365-2184.1970.tb00347.x Friedenstein AJ, Chailakhjan RK, Lalykina KS. The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet. 1970;3(4):393–403. 5523063 10.1111/j.1365-2184.1970.tb00347.x

55. Hoang DM Pham PT Bach TQ Ngo ATL Nguyen QT Phan TTK Stem cell-based therapy for human diseases Signal Transduct Target Ther 2022 7 1 272 35933430 10.1038/s41392-022-01134-4 PMC9357075 Hoang DM, Pham PT, Bach TQ, Ngo ATL, Nguyen QT, Phan TTK, et al. Stem cell-based therapy for human diseases. Signal Transduct Target Ther. 2022;7(1):272. 35933430 10.1038/s41392-022-01134-4 PMC9357075

56. Wu Z Su Y Li J Liu X Liu Y Zhao L Induced pluripotent stem cell-derived mesenchymal stem cells: whether they can become new stars of cell therapy Stem Cell Res Ther 2024 15 1 367 39415276 10.1186/s13287-024-03968-x PMC11484330 Wu Z, Su Y, Li J, Liu X, Liu Y, Zhao L, et al. Induced pluripotent stem cell-derived mesenchymal stem cells: whether they can become new stars of cell therapy. Stem Cell Res Ther. 2024;15(1):367. 39415276 10.1186/s13287-024-03968-x PMC11484330

57. Wang Q Yang Q Wang Z Tong H Ma L Zhang Y Comparative analysis of human mesenchymal stem cells from fetal-bone marrow, adipose tissue, and Warton’s jelly as sources of cell immunomodulatory therapy Hum Vaccin Immunother 2016 12 1 85 96 26186552 10.1080/21645515.2015.1030549 PMC4962749 Wang Q, Yang Q, Wang Z, Tong H, Ma L, Zhang Y, et al. Comparative analysis of human mesenchymal stem cells from fetal-bone marrow, adipose tissue, and Warton’s jelly as sources of cell immunomodulatory therapy. Hum Vaccin Immunother. 2016;12(1):85–96. 26186552 10.1080/21645515.2015.1030549 PMC4962749

58. Poomani MS Regurajan R Perumal R Ramachandran A Mariappan I Muthan K Differentiation of placenta-derived MSCs cultured in human platelet lysate: a xenofree supplement 3 Biotech 2024 14 4 116 38524240 10.1007/s13205-024-03966-z PMC10959853 Poomani MS, Regurajan R, Perumal R, Ramachandran A, Mariappan I, Muthan K, et al. Differentiation of placenta-derived MSCs cultured in human platelet lysate: a xenofree supplement. 3 Biotech. 2024;14(4):116. 38524240 10.1007/s13205-024-03966-z PMC10959853

59. Sharpe PT Dental mesenchymal stem cells Development 2016 143 13 2273 80 27381225 10.1242/dev.134189 Sharpe PT. Dental mesenchymal stem cells. Development. 2016;143(13):2273–80. 27381225 10.1242/dev.134189

60. Joerger-Messerli MS Marx C Oppliger B Mueller M Surbek DV Schoeberlein A Mesenchymal stem cells from Wharton’s jelly and amniotic fluid Best Pract Res Clin Obstet Gynaecol 2016 31 30 44 26482184 10.1016/j.bpobgyn.2015.07.006 Joerger-Messerli MS, Marx C, Oppliger B, Mueller M, Surbek DV, Schoeberlein A. Mesenchymal stem cells from Wharton’s jelly and amniotic fluid. Best Pract Res Clin Obstet Gynaecol. 2016;31:30–44. 26482184 10.1016/j.bpobgyn.2015.07.006

61. El Omar R Beroud J Stoltz J-F Menu P Velot E Decot V Umbilical cord mesenchymal stem cells: the new gold standard for mesenchymal stem cell-based therapies? Tissue Engineering Part B: Reviews 2014 20 5 523 44 24552279 10.1089/ten.TEB.2013.0664 El Omar R, Beroud J, Stoltz J-F, Menu P, Velot E, Decot V. Umbilical cord mesenchymal stem cells: the new gold standard for mesenchymal stem cell-based therapies? Tissue Engineering Part B: Reviews. 2014;20(5):523–44. 24552279 10.1089/ten.TEB.2013.0664

62. Dominici M Le Blanc K Mueller I Slaper-Cortenbach I Marini F Krause D Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement Cytotherapy 2006 8 4 315 7 16923606 10.1080/14653240600855905 Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy. 2006;8(4):315–7. 16923606 10.1080/14653240600855905

63. Topoluk N Hawkins R Tokish J Mercuri J Amniotic mesenchymal stromal cells exhibit preferential osteogenic and chondrogenic differentiation and enhanced matrix production compared with adipose mesenchymal stromal cells Am J Sports Med 2017 45 11 2637 46 28541092 10.1177/0363546517706138 PMC5832055 Topoluk N, Hawkins R, Tokish J, Mercuri J. Amniotic mesenchymal stromal cells exhibit preferential osteogenic and chondrogenic differentiation and enhanced matrix production compared with adipose mesenchymal stromal cells. Am J Sports Med. 2017;45(11):2637–46. 28541092 10.1177/0363546517706138 PMC5832055

64. Yoshimura H Muneta T Nimura A Yokoyama A Koga H Sekiya I Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle Cell Tissue Res 2007 327 3 449 62 17053900 10.1007/s00441-006-0308-z Yoshimura H, Muneta T, Nimura A, Yokoyama A, Koga H, Sekiya I. Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Res. 2007;327(3):449–62. 17053900 10.1007/s00441-006-0308-z

65. Chen S Liang B Xu J Unveiling heterogeneity in MSCs: exploring marker-based strategies for defining MSC subpopulations J Transl Med 2024 22 1 459 38750573 10.1186/s12967-024-05294-5 PMC11094970 Chen S, Liang B, Xu J. Unveiling heterogeneity in MSCs: exploring marker-based strategies for defining MSC subpopulations. J Transl Med. 2024;22(1):459. 38750573 10.1186/s12967-024-05294-5 PMC11094970

66. Li Q Hou H Li M Yu X Zuo H Gao J CD73 + mesenchymal stem cells ameliorate myocardial infarction by promoting angiogenesis Front Cell Dev Biol 2021 9 637239 34055772 10.3389/fcell.2021.637239 PMC8152667 Li Q, Hou H, Li M, Yu X, Zuo H, Gao J, et al. CD73 + mesenchymal stem cells ameliorate myocardial infarction by promoting angiogenesis. Front Cell Dev Biol. 2021;9:637239. 34055772 10.3389/fcell.2021.637239 PMC8152667

67. Smith RJP Faroni A Barrow JR Soul J Reid AJ The angiogenic potential of CD271 + human adipose tissue-derived mesenchymal stem cells Stem Cell Res Ther 2021 12 1 160 33653407 10.1186/s13287-021-02177-0 PMC7927269 Smith RJP, Faroni A, Barrow JR, Soul J, Reid AJ. The angiogenic potential of CD271 + human adipose tissue-derived mesenchymal stem cells. Stem Cell Res Ther. 2021;12(1):160. 33653407 10.1186/s13287-021-02177-0 PMC7927269

68. Li L Yang L Chen X Chen X Diao L Zeng Y TNFAIP6 defines the MSC subpopulation with enhanced immune suppression activities Stem Cell Res Ther 2022 13 1 479 36153571 10.1186/s13287-022-03176-5 PMC9509641 Li L, Yang L, Chen X, Chen X, Diao L, Zeng Y, et al. TNFAIP6 defines the MSC subpopulation with enhanced immune suppression activities. Stem Cell Res Ther. 2022;13(1):479. 36153571 10.1186/s13287-022-03176-5 PMC9509641

69. Hu D Lai J Chen Q Bai L New advances of NG2-expressing cell subset in marrow mesenchymal stem cells as novel therapeutic tools for liver fibrosis/cirrhosis Stem Cell Res Ther 2024 15 1 199 38971781 10.1186/s13287-024-03817-x PMC11227708 Hu D, Lai J, Chen Q, Bai L. New advances of NG2-expressing cell subset in marrow mesenchymal stem cells as novel therapeutic tools for liver fibrosis/cirrhosis. Stem Cell Res Ther. 2024;15(1):199. 38971781 10.1186/s13287-024-03817-x PMC11227708

70. Jia Y Wang A Zhao B Wang C Su R Zhang B An optimized method for obtaining clinical-grade specific cell subpopulations from human umbilical cord-derived mesenchymal stem cells Cell Prolif 2022 55 10 e13300 35768999 10.1111/cpr.13300 PMC9528761 Jia Y, Wang A, Zhao B, Wang C, Su R, Zhang B, et al. An optimized method for obtaining clinical-grade specific cell subpopulations from human umbilical cord-derived mesenchymal stem cells. Cell Prolif. 2022;55(10):e13300. 35768999 10.1111/cpr.13300 PMC9528761

71. Song J Ma Q Li Y Wang X Chen S Liang B CD317 + MSCs expanded with chemically defined media have enhanced immunological anti-inflammatory activities Stem Cell Res Ther 2024 15 1 2 38169422 10.1186/s13287-023-03618-8 PMC10763464 Song J, Ma Q, Li Y, Wang X, Chen S, Liang B, et al. CD317 + MSCs expanded with chemically defined media have enhanced immunological anti-inflammatory activities. Stem Cell Res Ther. 2024;15(1):2. 38169422 10.1186/s13287-023-03618-8 PMC10763464

72. Wu C-C Liu F-L Sytwu H-K Tsai C-Y Chang D-M CD146 + mesenchymal stem cells display greater therapeutic potential than CD146- cells for treating collagen-induced arthritis in mice Stem Cell Res Ther 2016 7 23 26841872 10.1186/s13287-016-0285-4 PMC4741021 Wu C-C, Liu F-L, Sytwu H-K, Tsai C-Y, Chang D-M. CD146 + mesenchymal stem cells display greater therapeutic potential than CD146- cells for treating collagen-induced arthritis in mice. Stem Cell Res Ther. 2016;7:23. 26841872 10.1186/s13287-016-0285-4 PMC4741021

73. Zhang C Chen X-H Zhang X Gao L Kong P-Y Peng X-g Human umbilical cord blood-derived stromal cells, a new resource in the suppression of acute graft-versus-host disease in haploidentical stem cell transplantation in sublethally irradiated mice J Biol Chem 2011 286 15 13723 32 21349838 10.1074/jbc.M110.144691 PMC3075716 Zhang C, Chen X-H, Zhang X, Gao L, Kong P-Y, Peng X-g, et al. Human umbilical cord blood-derived stromal cells, a new resource in the suppression of acute graft-versus-host disease in haploidentical stem cell transplantation in sublethally irradiated mice. J Biol Chem. 2011;286(15):13723–32. 21349838 10.1074/jbc.M110.144691 PMC3075716

74. Gao L Zhang Y Hu B Liu J Kong P Lou S Phase II multicenter, randomized, double-blind controlled study of efficacy and safety of umbilical cord-derived mesenchymal stromal cells in the prophylaxis of chronic graft-versus-host disease after HLA-haploidentical stem-cell transplantation J Clin Oncol 2016 34 24 2843 50 27400949 10.1200/JCO.2015.65.3642 Gao L, Zhang Y, Hu B, Liu J, Kong P, Lou S, et al. Phase II multicenter, randomized, double-blind controlled study of efficacy and safety of umbilical cord-derived mesenchymal stromal cells in the prophylaxis of chronic graft-versus-host disease after HLA-haploidentical stem-cell transplantation. J Clin Oncol. 2016;34(24):2843–50. 27400949 10.1200/JCO.2015.65.3642

75. Zhang C, Chen X-H, Zhang X, Gao L, Gao L, Kong P-Y, et al. Regulation of acute graft-versus-host disease by human umbilical cord blood derived stromal cells in haploidentical stem cell transplantation in mice through very late activation antigen-4. Clin Immunol. 2011;101. 10.1016/j.clim.2011.01.004. 10.1016/j.clim.2011.01.004 21310663

76. Zhang C Zhang X Chen X-H Hypothesis: human umbilical cord blood-derived stromal cells regulate the Foxp3 expression of regulatory T cells through the TGF-β1/Smad3 pathway Cell Biochem Biophys 2012 62 3 463 6 22161558 10.1007/s12013-011-9328-8 Zhang C, Zhang X, Chen X-H. Hypothesis: human umbilical cord blood-derived stromal cells regulate the Foxp3 expression of regulatory T cells through the TGF-β1/Smad3 pathway. Cell Biochem Biophys. 2012;62(3):463–6. 22161558 10.1007/s12013-011-9328-8

77. Gurrea-Rubio M Lin F Wicha MS Mao-Draayer Y Fox DA Ligands of CD6: roles in the pathogenesis and treatment of cancer Front Immunol 2024 15 1528478 39840036 10.3389/fimmu.2024.1528478 PMC11747410 Gurrea-Rubio M, Lin F, Wicha MS, Mao-Draayer Y, Fox DA. Ligands of CD6: roles in the pathogenesis and treatment of cancer. Front Immunol. 2024;15:1528478. 39840036 10.3389/fimmu.2024.1528478 PMC11747410

78. Towers R Trombello L Fusenig M Tunger A Baumann A-L Savoldelli R Bone marrow-derived mesenchymal stromal cells obstruct AML-targeting CD8 + clonal effector and CAR T-cell function while promoting a senescence-associated phenotype Cancer Immunol Immunother 2024 73 1 8 38231344 10.1007/s00262-023-03594-1 PMC10794426 Towers R, Trombello L, Fusenig M, Tunger A, Baumann A-L, Savoldelli R, et al. Bone marrow-derived mesenchymal stromal cells obstruct AML-targeting CD8 + clonal effector and CAR T-cell function while promoting a senescence-associated phenotype. Cancer Immunol Immunother. 2024;73(1):8. 38231344 10.1007/s00262-023-03594-1 PMC10794426

79. Zhang R Liu Q Zhou S He H Zhao M Ma W Mesenchymal stem cell suppresses the efficacy of CAR-T toward killing lymphoma cells by modulating the microenvironment through stanniocalcin-1 Elife 2023 12 e82934 10.7554/eLife.82934 36779699 PMC10019890 Zhang R, Liu Q, Zhou S, He H, Zhao M, Ma W. Mesenchymal stem cell suppresses the efficacy of CAR-T toward killing lymphoma cells by modulating the microenvironment through stanniocalcin-1. Elife. 2023;12:e82934. 10.7554/eLife.82934 36779699 10.7554/eLife.82934 PMC10019890

80. Holthof LC van der Schans JJ Katsarou A Poels R Gelderloos AT Drent E Bone marrow mesenchymal stromal cells can render multiple myeloma cells resistant to cytotoxic machinery of CAR T cells through inhibition of apoptosis Clin Cancer Res 2021 27 13 3793 803 33883175 10.1158/1078-0432.CCR-20-2188 Holthof LC, van der Schans JJ, Katsarou A, Poels R, Gelderloos AT, Drent E, et al. Bone marrow mesenchymal stromal cells can render multiple myeloma cells resistant to cytotoxic machinery of CAR T cells through inhibition of apoptosis. Clin Cancer Res. 2021;27(13):3793–803. 33883175 10.1158/1078-0432.CCR-20-2188

81. Holthof LC Stikvoort A van der Horst HJ Gelderloos AT Poels R Li F Bone marrow mesenchymal stromal cell-mediated resistance in multiple myeloma against NK cells can be overcome by introduction of CD38-CAR or TRAIL-variant Hemasphere 2021 5 5 e561 33898931 10.1097/HS9.0000000000000561 PMC8061681 Holthof LC, Stikvoort A, van der Horst HJ, Gelderloos AT, Poels R, Li F, et al. Bone marrow mesenchymal stromal cell-mediated resistance in multiple myeloma against NK cells can be overcome by introduction of CD38-CAR or TRAIL-variant. Hemasphere. 2021;5(5):e561. 33898931 10.1097/HS9.0000000000000561 PMC8061681

82. Court AC Parra-Crisóstomo E Castro-Córdova P Abdo L Aragão EAA Lorca R Survival advantage of native and engineered T cells is acquired by mitochondrial transfer from mesenchymal stem cells J Transl Med 2024 22 1 868 39334383 10.1186/s12967-024-05627-4 PMC11429672 Court AC, Parra-Crisóstomo E, Castro-Córdova P, Abdo L, Aragão EAA, Lorca R, et al. Survival advantage of native and engineered T cells is acquired by mitochondrial transfer from mesenchymal stem cells. J Transl Med. 2024;22(1):868. 39334383 10.1186/s12967-024-05627-4 PMC11429672

83. McKenna MK Englisch A Brenner B Smith T Hoyos V Suzuki M Mesenchymal stromal cell delivery of oncolytic immunotherapy improves CAR-T cell antitumor activity Mol Ther 2021 29 5 1808 20 33571680 10.1016/j.ymthe.2021.02.004 PMC8116608 McKenna MK, Englisch A, Brenner B, Smith T, Hoyos V, Suzuki M, et al. Mesenchymal stromal cell delivery of oncolytic immunotherapy improves CAR-T cell antitumor activity. Mol Ther. 2021;29(5):1808–20. 33571680 10.1016/j.ymthe.2021.02.004 PMC8116608

84. Hombach AA Geumann U Günther C Hermann FG Abken H IL7-IL12 engineered mesenchymal stem cells (MSCs) improve a CAR T cell attack against colorectal cancer cells Cells 2020 9 4 873 10.3390/cells9040873 32260097 PMC7226757 Hombach AA, Geumann U, Günther C, Hermann FG, Abken H. IL7-IL12 engineered mesenchymal stem cells (MSCs) improve a CAR T cell attack against colorectal cancer cells. Cells. 2020;9(4):873. 10.3390/cells9040873 32260097 10.3390/cells9040873 PMC7226757

85. Zanetti SR Romecin PA Vinyoles M Juan M Fuster JL Cámos M Bone marrow MSC from pediatric patients with B-ALL highly immunosuppress T-cell responses but do not compromise CD19-CAR T-cell activity J Immunother Cancer 2020 8 2 e001419 10.1136/jitc-2020-001419 32868394 PMC7462245 Zanetti SR, Romecin PA, Vinyoles M, Juan M, Fuster JL, Cámos M, et al. Bone marrow MSC from pediatric patients with B-ALL highly immunosuppress T-cell responses but do not compromise CD19-CAR T-cell activity. J Immunother Cancer. 2020;8(2):e001419. 10.1136/jitc-2020-001419 32868394 10.1136/jitc-2020-001419 PMC7462245

86. Xia Y Wang L Shen X Xu Y Xu W Li J Mesenchymal stem cell infusion for enhancing hematopoietic recovery and addressing cytopenias in CAR-T cell therapy Stem Cell Res Ther 2024 15 1 333 39334276 10.1186/s13287-024-03941-8 PMC11437967 Xia Y, Wang L, Shen X, Xu Y, Xu W, Li J, et al. Mesenchymal stem cell infusion for enhancing hematopoietic recovery and addressing cytopenias in CAR-T cell therapy. Stem Cell Res Ther. 2024;15(1):333. 39334276 10.1186/s13287-024-03941-8 PMC11437967

87. Hazrati A Malekpour K Khorramdelazad H Rajaei S Hashemi SM Therapeutic and immunomodulatory potentials of mesenchymal stromal/stem cells and immune checkpoints related molecules Biomark Res 2024 12 1 35 38515166 10.1186/s40364-024-00580-2 PMC10958918 Hazrati A, Malekpour K, Khorramdelazad H, Rajaei S, Hashemi SM. Therapeutic and immunomodulatory potentials of mesenchymal stromal/stem cells and immune checkpoints related molecules. Biomark Res. 2024;12(1):35. 38515166 10.1186/s40364-024-00580-2 PMC10958918

88. Zhou J Shi Y Mesenchymal stem/stromal cells (MSCs): origin, immune regulation, and clinical applications Cell Mol Immunol 2023 20 6 555 7 37225837 10.1038/s41423-023-01034-9 PMC10229593 Zhou J, Shi Y. Mesenchymal stem/stromal cells (MSCs): origin, immune regulation, and clinical applications. Cell Mol Immunol. 2023;20(6):555–7. 37225837 10.1038/s41423-023-01034-9 PMC10229593

89. Zhou Y Cheng L Liu L Li X NK cells are never alone: crosstalk and communication in tumour microenvironments Mol Cancer 2023 22 1 34 36797782 10.1186/s12943-023-01737-7 PMC9933398 Zhou Y, Cheng L, Liu L, Li X. NK cells are never alone: crosstalk and communication in tumour microenvironments. Mol Cancer. 2023;22(1):34. 36797782 10.1186/s12943-023-01737-7 PMC9933398

90. Salami F Tavassoli A Mehrzad J Parham A Immunomodulatory effects of mesenchymal stem cells on leukocytes with emphasis on neutrophils Immunobiology 2018 223 12 786 91 30119931 10.1016/j.imbio.2018.08.002 Salami F, Tavassoli A, Mehrzad J, Parham A. Immunomodulatory effects of mesenchymal stem cells on leukocytes with emphasis on neutrophils. Immunobiology. 2018;223(12):786–91. 30119931 10.1016/j.imbio.2018.08.002

91. Hezam K Wang C Fu E Zhou M Liu Y Wang H Superior protective effects of PGE2 priming mesenchymal stem cells against LPS-induced acute lung injury (ALI) through macrophage immunomodulation Stem Cell Res Ther 2023 14 1 48 36949464 10.1186/s13287-023-03277-9 PMC10032272 Hezam K, Wang C, Fu E, Zhou M, Liu Y, Wang H, et al. Superior protective effects of PGE2 priming mesenchymal stem cells against LPS-induced acute lung injury (ALI) through macrophage immunomodulation. Stem Cell Res Ther. 2023;14(1):48. 36949464 10.1186/s13287-023-03277-9 PMC10032272

92. Wang Y Chang Y-J Chen J Han M Hu J Hu J Consensus on the monitoring, treatment, and prevention of leukaemia relapse after allogeneic haematopoietic stem cell transplantation in China: 2024 update Cancer Lett 2024 605 217264 39332587 10.1016/j.canlet.2024.217264 Wang Y, Chang Y-J, Chen J, Han M, Hu J, Hu J, et al. Consensus on the monitoring, treatment, and prevention of leukaemia relapse after allogeneic haematopoietic stem cell transplantation in China: 2024 update. Cancer Lett. 2024;605:217264. 39332587 10.1016/j.canlet.2024.217264

93. Zhang X-H Chen J Han M-Z Huang H Jiang E-L Jiang M The consensus from the Chinese society of hematology on indications, conditioning regimens and donor selection for allogeneic hematopoietic stem cell transplantation: 2021 update J Hematol Oncol 2021 14 1 145 34526099 10.1186/s13045-021-01159-2 PMC8441240 Zhang X-H, Chen J, Han M-Z, Huang H, Jiang E-L, Jiang M, et al. The consensus from the Chinese society of hematology on indications, conditioning regimens and donor selection for allogeneic hematopoietic stem cell transplantation: 2021 update. J Hematol Oncol. 2021;14(1):145. 34526099 10.1186/s13045-021-01159-2 PMC8441240

94. Huang R Wang X Zhang X Unity brings strength: combination of CAR-T cell therapy and HSCT Cancer Lett 2022 549 215721 35537573 10.1016/j.canlet.2022.215721 Huang R, Wang X, Zhang X. Unity brings strength: combination of CAR-T cell therapy and HSCT. Cancer Lett. 2022;549:215721. 35537573 10.1016/j.canlet.2022.215721

95. Huang R Chen T Wang S Wang J Su Y Liu J Mesenchymal stem cells for prophylaxis of chronic graft-vs-host disease after haploidentical hematopoietic stem cell transplant: an open-label randomized clinical trial JAMA Oncol 2024 10 2 220 6 38153755 10.1001/jamaoncol.2023.5757 PMC10870190 Huang R, Chen T, Wang S, Wang J, Su Y, Liu J, et al. Mesenchymal stem cells for prophylaxis of chronic graft-vs-host disease after haploidentical hematopoietic stem cell transplant: an open-label randomized clinical trial. JAMA Oncol. 2024;10(2):220–6. 38153755 10.1001/jamaoncol.2023.5757 PMC10870190

96. Zhao K Lin R Fan Z Chen X Wang Y Huang F Mesenchymal stromal cells plus basiliximab, calcineurin inhibitor as treatment of steroid-resistant acute graft-versus-host disease: a multicenter, randomized, phase 3, open-label trial J Hematol Oncol 2022 15 1 22 35255929 10.1186/s13045-022-01240-4 PMC8900437 Zhao K, Lin R, Fan Z, Chen X, Wang Y, Huang F, et al. Mesenchymal stromal cells plus basiliximab, calcineurin inhibitor as treatment of steroid-resistant acute graft-versus-host disease: a multicenter, randomized, phase 3, open-label trial. J Hematol Oncol. 2022;15(1):22. 35255929 10.1186/s13045-022-01240-4 PMC8900437

97. Kelly K Bloor AJC Griffin JE Radia R Yeung DT Rasko JEJ Two-year safety outcomes of iPS cell-derived mesenchymal stromal cells in acute steroid-resistant graft-versus-host disease Nat Med 2024 30 6 1556 8 38778211 10.1038/s41591-024-02990-z PMC11186752 Kelly K, Bloor AJC, Griffin JE, Radia R, Yeung DT, Rasko JEJ. Two-year safety outcomes of iPS cell-derived mesenchymal stromal cells in acute steroid-resistant graft-versus-host disease. Nat Med. 2024;30(6):1556–8. 38778211 10.1038/s41591-024-02990-z PMC11186752

98. Jiang E Qian K Wang L Yang D Shao Y Hu L Efficacy and safety of human umbilical cord-derived mesenchymal stem cells versus placebo added to second-line therapy in patients with steroid-refractory acute graft-versus-host disease: a multicentre, randomized, double-blind, phase 2 trial BMC Med 2024 22 1 555 39587570 10.1186/s12916-024-03782-5 PMC11590523 Jiang E, Qian K, Wang L, Yang D, Shao Y, Hu L, et al. Efficacy and safety of human umbilical cord-derived mesenchymal stem cells versus placebo added to second-line therapy in patients with steroid-refractory acute graft-versus-host disease: a multicentre, randomized, double-blind, phase 2 trial. BMC Med. 2024;22(1):555. 39587570 10.1186/s12916-024-03782-5 PMC11590523

99. Farge D Biard L Weil B Girault V Lansiaux P Munia I Allogeneic umbilical cord-derived mesenchymal stromal cells as treatment for systemic lupus erythematosus: a single-centre, open-label, dose-escalation, phase 1 study Lancet Rheumatol 2025 7 4 e261 73 39706212 10.1016/S2665-9913(24)00298-4 Farge D, Biard L, Weil B, Girault V, Lansiaux P, Munia I, et al. Allogeneic umbilical cord-derived mesenchymal stromal cells as treatment for systemic lupus erythematosus: a single-centre, open-label, dose-escalation, phase 1 study. Lancet Rheumatol. 2025;7(4):e261–73. 39706212 10.1016/S2665-9913(24)00298-4

100. Harris VK Stark J Williams A Roche M Malin M Kumar A Efficacy of intrathecal mesenchymal stem cell-neural progenitor therapy in progressive MS: results from a phase II, randomized, placebo-controlled clinical trial Stem Cell Res Ther 2024 15 1 151 38783390 10.1186/s13287-024-03765-6 PMC11119709 Harris VK, Stark J, Williams A, Roche M, Malin M, Kumar A, et al. Efficacy of intrathecal mesenchymal stem cell-neural progenitor therapy in progressive MS: results from a phase II, randomized, placebo-controlled clinical trial. Stem Cell Res Ther. 2024;15(1):151. 38783390 10.1186/s13287-024-03765-6 PMC11119709

101. Seo H-M Lew B-L Lee YW Son SW Park CO Park YL Phase 1/2 trials of human bone marrow-derived clonal mesenchymal stem cells for treatment of adults with moderate to severe atopic dermatitis J Allergy Clin Immunol 2024 154 4 965 73 38944393 10.1016/j.jaci.2024.06.013 Seo H-M, Lew B-L, Lee YW, Son SW, Park CO, Park YL, et al. Phase 1/2 trials of human bone marrow-derived clonal mesenchymal stem cells for treatment of adults with moderate to severe atopic dermatitis. J Allergy Clin Immunol. 2024;154(4):965–73. 38944393 10.1016/j.jaci.2024.06.013

102. Reenaers C Gillard RP Coimbra C Gillard RM Meunier P Lechanteur C Clinical and MRI evolution after local injection of bone marrow-derived mesenchymal stem cells in perianal fistulae in Crohn’s disease: results from a prospective monocentric study J Crohns Colitis 2023 17 5 728 37 36733215 10.1093/ecco-jcc/jjac192 Reenaers C, Gillard RP, Coimbra C, Gillard RM, Meunier P, Lechanteur C, et al. Clinical and MRI evolution after local injection of bone marrow-derived mesenchymal stem cells in perianal fistulae in Crohn’s disease: results from a prospective monocentric study. J Crohns Colitis. 2023;17(5):728–37. 36733215 10.1093/ecco-jcc/jjac192

103. Sirpilla O Sakemura RL Hefazi M Huynh TN Can I Girsch JH Mesenchymal stromal cells with chimaeric antigen receptors for enhanced immunosuppression Nat Biomed Eng 2024 8 4 443 60 38561490 10.1038/s41551-024-01195-6 PMC12080371 Sirpilla O, Sakemura RL, Hefazi M, Huynh TN, Can I, Girsch JH, et al. Mesenchymal stromal cells with chimaeric antigen receptors for enhanced immunosuppression. Nat Biomed Eng. 2024;8(4):443–60. 38561490 10.1038/s41551-024-01195-6 PMC12080371

104. Liu S-S Zhang C Zhang X Chen X-H Human umbilical cord blood-derived stromal cells: a new source of stromal cells in hematopoietic stem cell transplantation Crit Rev Oncol Hematol 2014 90 2 93 8 24411588 10.1016/j.critrevonc.2013.12.002 Liu S-S, Zhang C, Zhang X, Chen X-H. Human umbilical cord blood-derived stromal cells: a new source of stromal cells in hematopoietic stem cell transplantation. Crit Rev Oncol Hematol. 2014;90(2):93–8. 24411588 10.1016/j.critrevonc.2013.12.002

105. Zhang C Zhang X Chen X-H Granulocyte-colony stimulating factor-mobilized mesenchymal stem cells: a new resource for rapid engraftment in hematopoietic stem cell transplantation Med Hypotheses 2011 76 2 241 3 21050671 10.1016/j.mehy.2010.10.008 Zhang C, Zhang X, Chen X-H. Granulocyte-colony stimulating factor-mobilized mesenchymal stem cells: a new resource for rapid engraftment in hematopoietic stem cell transplantation. Med Hypotheses. 2011;76(2):241–3. 21050671 10.1016/j.mehy.2010.10.008

106. Tai T-S Chen Y-H Yao C-L Lin J-H Yang Y-S Shi J-W Cellular sentinels: empowering survival and immune defense in hematopoietic stem cell transplantation through mesenchymal stem cells and T lymphocytes BMC Med 2025 23 1 164 40102849 10.1186/s12916-025-03987-2 PMC11921582 Tai T-S, Chen Y-H, Yao C-L, Lin J-H, Yang Y-S, Shi J-W, et al. Cellular sentinels: empowering survival and immune defense in hematopoietic stem cell transplantation through mesenchymal stem cells and T lymphocytes. BMC Med. 2025;23(1):164. 40102849 10.1186/s12916-025-03987-2 PMC11921582

107. Lin T Yang Y Chen X A review of the application of mesenchymal stem cells in the field of hematopoietic stem cell transplantation Eur J Med Res 2023 28 1 268 37550742 10.1186/s40001-023-01244-x PMC10405442 Lin T, Yang Y, Chen X. A review of the application of mesenchymal stem cells in the field of hematopoietic stem cell transplantation. Eur J Med Res. 2023;28(1):268. 37550742 10.1186/s40001-023-01244-x PMC10405442

108. Lan Y Liu F Chang L Liu L Zhang Y Yi M Combination of umbilical cord mesenchymal stem cells and standard immunosuppressive regimen for pediatric patients with severe aplastic anemia BMC Pediatr 2021 21 1 102 33639900 10.1186/s12887-021-02562-x PMC7912947 Lan Y, Liu F, Chang L, Liu L, Zhang Y, Yi M, et al. Combination of umbilical cord mesenchymal stem cells and standard immunosuppressive regimen for pediatric patients with severe aplastic anemia. BMC Pediatr. 2021;21(1):102. 33639900 10.1186/s12887-021-02562-x PMC7912947

109. Chen Y Xu Y Chi Y Sun T Gao Y Dou X Efficacy and safety of human umbilical cord-derived mesenchymal stem cells in the treatment of refractory immune thrombocytopenia: a prospective, single arm, phase I trial Signal Transduct Target Ther 2024 9 1 102 38653983 10.1038/s41392-024-01793-5 PMC11039759 Chen Y, Xu Y, Chi Y, Sun T, Gao Y, Dou X, et al. Efficacy and safety of human umbilical cord-derived mesenchymal stem cells in the treatment of refractory immune thrombocytopenia: a prospective, single arm, phase I trial. Signal Transduct Target Ther. 2024;9(1):102. 38653983 10.1038/s41392-024-01793-5 PMC11039759

110. Xu W Yang Y Li N Hua J Interaction between mesenchymal stem cells and immune cells during bone injury repair Int J Mol Sci 2023 24 19 10.3390/ijms241914484 PMC10572976 37833933 Xu W, Yang Y, Li N, Hua J. Interaction between mesenchymal stem cells and immune cells during bone injury repair. Int J Mol Sci. 2023;24:19. 10.3390/ijms241914484 PMC10572976 37833933

111. Wu C Huang Z Chen J Li N Cai Y Chen J Efficiently directing differentiation and homing of mesenchymal stem cells to boost cartilage repair in osteoarthritis via a nanoparticle and peptide dual-engineering strategy Biomaterials 2025 312 122720 39084098 10.1016/j.biomaterials.2024.122720 Wu C, Huang Z, Chen J, Li N, Cai Y, Chen J, et al. Efficiently directing differentiation and homing of mesenchymal stem cells to boost cartilage repair in osteoarthritis via a nanoparticle and peptide dual-engineering strategy. Biomaterials. 2025;312:122720. 39084098 10.1016/j.biomaterials.2024.122720

112. Zhang Y Ju W Zhang H Mengyun L Shen W Chen X Mechanisms and therapeutic prospects of mesenchymal stem cells-derived exosomes for tendinopathy Stem Cell Res Ther 2023 14 1 307 37880763 10.1186/s13287-023-03431-3 PMC10601253 Zhang Y, Ju W, Zhang H, Mengyun L, Shen W, Chen X. Mechanisms and therapeutic prospects of mesenchymal stem cells-derived exosomes for tendinopathy. Stem Cell Res Ther. 2023;14(1):307. 37880763 10.1186/s13287-023-03431-3 PMC10601253

113. Yadav P Singh SK Rajput S Allawadhi P Khurana A Weiskirchen R Therapeutic potential of stem cells in regeneration of liver in chronic liver diseases: current perspectives and future challenges Pharmacol Ther 2024 253 108563 38013053 10.1016/j.pharmthera.2023.108563 Yadav P, Singh SK, Rajput S, Allawadhi P, Khurana A, Weiskirchen R, et al. Therapeutic potential of stem cells in regeneration of liver in chronic liver diseases: current perspectives and future challenges. Pharmacol Ther. 2024;253:108563. 38013053 10.1016/j.pharmthera.2023.108563

114. Yao S Pang M Wang Y Wang X Lin Y Lv Y Mesenchymal stem cell attenuates spinal cord injury by inhibiting mitochondrial quality control-associated neuronal ferroptosis Redox Biol 2023 67 102871 37699320 10.1016/j.redox.2023.102871 PMC10506061 Yao S, Pang M, Wang Y, Wang X, Lin Y, Lv Y, et al. Mesenchymal stem cell attenuates spinal cord injury by inhibiting mitochondrial quality control-associated neuronal ferroptosis. Redox Biol. 2023;67:102871. 37699320 10.1016/j.redox.2023.102871 PMC10506061

115. Sajadi S Khadembashiri MA Raissi G Khadembashiri MM Mansouri K Hadizadeh-Kharazi H The role of adipose-derived stem cells in knee osteoarthritis treatment: insights from a triple-blind clinical study Stem Cell Res Ther 2025 16 1 242 40369655 10.1186/s13287-025-04233-5 PMC12079929 Sajadi S, Khadembashiri MA, Raissi G, Khadembashiri MM, Mansouri K, Hadizadeh-Kharazi H, et al. The role of adipose-derived stem cells in knee osteoarthritis treatment: insights from a triple-blind clinical study. Stem Cell Res Ther. 2025;16(1):242. 40369655 10.1186/s13287-025-04233-5 PMC12079929

116. Yan W Xia Y Zhao H Xu X Ma X Tao L Stem cell-based therapy in cardiac repair after myocardial infarction: promise, challenges, and future directions J Mol Cell Cardiol 2024 188 1 14 10.1016/j.yjmcc.2023.12.009 38246086 Yan W, Xia Y, Zhao H, Xu X, Ma X, Tao L. Stem cell-based therapy in cardiac repair after myocardial infarction: promise, challenges, and future directions. J Mol Cell Cardiol. 2024;188:1–14. 10.1016/j.yjmcc.2023.12.009 38246086 10.1016/j.yjmcc.2023.12.009

117. Attar A Farjoud Kouhanjani M Hessami K Vosough M Kojuri J Ramzi M Effect of once versus twice intracoronary injection of allogeneic-derived mesenchymal stromal cells after acute myocardial infarction: BOOSTER-TAHA7 randomized clinical trial Stem Cell Res Ther 2023 14 1 264 37740221 10.1186/s13287-023-03495-1 PMC10517503 Attar A, Farjoud Kouhanjani M, Hessami K, Vosough M, Kojuri J, Ramzi M, et al. Effect of once versus twice intracoronary injection of allogeneic-derived mesenchymal stromal cells after acute myocardial infarction: BOOSTER-TAHA7 randomized clinical trial. Stem Cell Res Ther. 2023;14(1):264. 37740221 10.1186/s13287-023-03495-1 PMC10517503

118. Qayyum AA van Klarenbosch B Frljak S Cerar A Poglajen G Traxler-Weidenauer D Effect of allogeneic adipose tissue-derived mesenchymal stromal cell treatment in chronic ischaemic heart failure with reduced ejection fraction - the SCIENCE trial Eur J Heart Fail 2023 25 4 576 87 36644821 10.1002/ejhf.2772 Qayyum AA, van Klarenbosch B, Frljak S, Cerar A, Poglajen G, Traxler-Weidenauer D, et al. Effect of allogeneic adipose tissue-derived mesenchymal stromal cell treatment in chronic ischaemic heart failure with reduced ejection fraction - the SCIENCE trial. Eur J Heart Fail. 2023;25(4):576–87. 36644821 10.1002/ejhf.2772

119. Ding J-Y Chen M-J Wu L-F Shu G-F Fang S-J Li Z-Y Mesenchymal stem cell-derived extracellular vesicles in skin wound healing: roles, opportunities and challenges Mil Med Res 2023 10 1 36 37587531 10.1186/s40779-023-00472-w PMC10433599 Ding J-Y, Chen M-J, Wu L-F, Shu G-F, Fang S-J, Li Z-Y, et al. Mesenchymal stem cell-derived extracellular vesicles in skin wound healing: roles, opportunities and challenges. Mil Med Res. 2023;10(1):36. 37587531 10.1186/s40779-023-00472-w PMC10433599

120. Shirbaghaee Z Heidari Keshel S Rasouli M Valizadeh M Hashemi Nazari SS Hassani M Report of a phase 1 clinical trial for safety assessment of human placental mesenchymal stem cells therapy in patients with critical limb ischemia (CLI) Stem Cell Res Ther 2023 14 1 174 37408043 10.1186/s13287-023-03390-9 PMC10324209 Shirbaghaee Z, Heidari Keshel S, Rasouli M, Valizadeh M, Hashemi Nazari SS, Hassani M, et al. Report of a phase 1 clinical trial for safety assessment of human placental mesenchymal stem cells therapy in patients with critical limb ischemia (CLI). Stem Cell Res Ther. 2023;14(1):174. 37408043 10.1186/s13287-023-03390-9 PMC10324209

121. Rash BG Ramdas KN Agafonova N Naioti E McClain-Moss L Zainul Z Allogeneic mesenchymal stem cell therapy with laromestrocel in mild Alzheimer’s disease: a randomized controlled phase 2a trial Nat Med 2025 31 4 1257 66 40065171 10.1038/s41591-025-03559-0 PMC12003194 Rash BG, Ramdas KN, Agafonova N, Naioti E, McClain-Moss L, Zainul Z, et al. Allogeneic mesenchymal stem cell therapy with laromestrocel in mild Alzheimer’s disease: a randomized controlled phase 2a trial. Nat Med. 2025;31(4):1257–66. 40065171 10.1038/s41591-025-03559-0 PMC12003194

122. Zhuo Y Li W-S Lu W Li X Ge L-T Huang Y TGF-β1 mediates hypoxia-preconditioned olfactory mucosa mesenchymal stem cells improved neural functional recovery in parkinson’s disease models and patients Mil Med Res 2024 11 1 48 39034405 10.1186/s40779-024-00550-7 PMC11265117 Zhuo Y, Li W-S, Lu W, Li X, Ge L-T, Huang Y, et al. TGF-β1 mediates hypoxia-preconditioned olfactory mucosa mesenchymal stem cells improved neural functional recovery in parkinson’s disease models and patients. Mil Med Res. 2024;11(1):48. 39034405 10.1186/s40779-024-00550-7 PMC11265117

123. Giovannelli L Bari E Jommi C Tartara F Armocida D Garbossa D Mesenchymal stem cell secretome and extracellular vesicles for neurodegenerative diseases: risk-benefit profile and next steps for the market access Bioact Mater 2023 29 16 35 37456581 10.1016/j.bioactmat.2023.06.013 PMC10338239 Giovannelli L, Bari E, Jommi C, Tartara F, Armocida D, Garbossa D, et al. Mesenchymal stem cell secretome and extracellular vesicles for neurodegenerative diseases: risk-benefit profile and next steps for the market access. Bioact Mater. 2023;29:16–35. 37456581 10.1016/j.bioactmat.2023.06.013 PMC10338239

124. Lee J Chang WH Chung J-W Kim SJ Kim S-K Lee JS Efficacy of intravenous mesenchymal stem cells for motor recovery after ischemic stroke: a neuroimaging study Stroke 2022 53 1 20 8 34583525 10.1161/STROKEAHA.121.034505 Lee J, Chang WH, Chung J-W, Kim SJ, Kim S-K, Lee JS, et al. Efficacy of intravenous mesenchymal stem cells for motor recovery after ischemic stroke: a neuroimaging study. Stroke. 2022;53(1):20–8. 34583525 10.1161/STROKEAHA.121.034505

125. Baak LM Wagenaar N van der Aa NE Groenendaal F Dudink J Tataranno ML Feasibility and safety of intranasally administered mesenchymal stromal cells after perinatal arterial ischaemic stroke in the Netherlands (PASSIoN): a first-in-human, open-label intervention study Lancet Neurol 2022 21 6 528 36 35568047 10.1016/S1474-4422(22)00117-X Baak LM, Wagenaar N, van der Aa NE, Groenendaal F, Dudink J, Tataranno ML, et al. Feasibility and safety of intranasally administered mesenchymal stromal cells after perinatal arterial ischaemic stroke in the Netherlands (PASSIoN): a first-in-human, open-label intervention study. Lancet Neurol. 2022;21(6):528–36. 35568047 10.1016/S1474-4422(22)00117-X

126. Durand NC Kim HG Patel VN Turnbull MT Siegel JL Hodge DO Mesenchymal stem cell therapy in acute intracerebral hemorrhage: a dose-escalation safety and tolerability trial Neurocrit Care 2024 41 1 59 69 38114796 10.1007/s12028-023-01897-w PMC11335835 Durand NC, Kim HG, Patel VN, Turnbull MT, Siegel JL, Hodge DO, et al. Mesenchymal stem cell therapy in acute intracerebral hemorrhage: a dose-escalation safety and tolerability trial. Neurocrit Care. 2024;41(1):59–69. 38114796 10.1007/s12028-023-01897-w PMC11335835

127. Okonkwo DO McAllister P Achrol AS Karasawa Y Kawabori M Cramer SC Mesenchymal stromal cell implants for chronic motor deficits after traumatic brain injury: post hoc analysis of a randomized trial Neurology 2024 103 7 e209797 39231380 10.1212/WNL.0000000000209797 PMC11373674 Okonkwo DO, McAllister P, Achrol AS, Karasawa Y, Kawabori M, Cramer SC, et al. Mesenchymal stromal cell implants for chronic motor deficits after traumatic brain injury: post hoc analysis of a randomized trial. Neurology. 2024;103(7):e209797. 39231380 10.1212/WNL.0000000000209797 PMC11373674

128. Pastor JC Pastor-Idoate S López-Paniagua M Para M Blazquez F Murgui E Intravitreal allogeneic mesenchymal stem cells: a non-randomized phase II clinical trial for acute non-arteritic optic neuropathy Stem Cell Res Ther 2023 14 1 261 37735668 10.1186/s13287-023-03500-7 PMC10512539 Pastor JC, Pastor-Idoate S, López-Paniagua M, Para M, Blazquez F, Murgui E, et al. Intravitreal allogeneic mesenchymal stem cells: a non-randomized phase II clinical trial for acute non-arteritic optic neuropathy. Stem Cell Res Ther. 2023;14(1):261. 37735668 10.1186/s13287-023-03500-7 PMC10512539

129. François S Usunier B Forgue-Lafitte M-E L’Homme B Benderitter M Douay L Mesenchymal stem cell administration attenuates colon cancer progression by modulating the immune component within the colorectal tumor microenvironment Stem Cells Transl Med 2019 8 3 285 300 30451398 10.1002/sctm.18-0117 PMC6392393 François S, Usunier B, Forgue-Lafitte M-E, L’Homme B, Benderitter M, Douay L, et al. Mesenchymal stem cell administration attenuates colon cancer progression by modulating the immune component within the colorectal tumor microenvironment. Stem Cells Transl Med. 2019;8(3):285–300. 30451398 10.1002/sctm.18-0117 PMC6392393

130. Vanegas N-DP Vernot J-P Loss of quiescence and self-renewal capacity of hematopoietic stem cell in an in vitro leukemic niche Exp Hematol Oncol 2017 6 2 28078190 10.1186/s40164-016-0062-1 PMC5223333 Vanegas N-DP, Vernot J-P. Loss of quiescence and self-renewal capacity of hematopoietic stem cell in an in vitro leukemic niche. Exp Hematol Oncol. 2017;6:2. 28078190 10.1186/s40164-016-0062-1 PMC5223333

131. Mirabdollahi M Haghjooyjavanmard S Sadeghi-Aliabadi H An anticancer effect of umbilical cord-derived mesenchymal stem cell secretome on the breast cancer cell line Cell Tissue Bank 2019 20 3 423 34 31338647 10.1007/s10561-019-09781-8 Mirabdollahi M, Haghjooyjavanmard S, Sadeghi-Aliabadi H. An anticancer effect of umbilical cord-derived mesenchymal stem cell secretome on the breast cancer cell line. Cell Tissue Bank. 2019;20(3):423–34. 31338647 10.1007/s10561-019-09781-8

132. Farahzadi R Fathi E Vandghanooni S Valipour B Cytokines secreted from bone marrow-derived mesenchymal stem cells promote apoptosis of CD34 + leukemic stem cells as anti-cancer therapy Regen Ther 2024 26 646 53 39281104 10.1016/j.reth.2024.08.008 PMC11401101 Farahzadi R, Fathi E, Vandghanooni S, Valipour B. Cytokines secreted from bone marrow-derived mesenchymal stem cells promote apoptosis of CD34 + leukemic stem cells as anti-cancer therapy. Regen Ther. 2024;26:646–53. 39281104 10.1016/j.reth.2024.08.008 PMC11401101

133. Fakiruddin KS Ghazalli N Lim MN Zakaria Z Abdullah S Mesenchymal stem cell expressing TRAIL as targeted therapy against sensitised tumour Int J Mol Sci 2018 19 8 2188 30060445 10.3390/ijms19082188 PMC6121609 Fakiruddin KS, Ghazalli N, Lim MN, Zakaria Z, Abdullah S. Mesenchymal stem cell expressing TRAIL as targeted therapy against sensitised tumour. Int J Mol Sci. 2018;19(8):2188. 30060445 10.3390/ijms19082188 PMC6121609

134. Stefani FR Eberstål S Vergani S Kristiansen TA Bengzon J Low-dose irradiated mesenchymal stromal cells break tumor defensive properties in vivo Int J Cancer 2018 143 9 2200 12 29752716 10.1002/ijc.31599 PMC6220775 Stefani FR, Eberstål S, Vergani S, Kristiansen TA, Bengzon J. Low-dose irradiated mesenchymal stromal cells break tumor defensive properties in vivo. Int J Cancer. 2018;143(9):2200–12. 29752716 10.1002/ijc.31599 PMC6220775

135. Zhou Y Li Y Wang H Sun H Su J Fan Y Mesenchymal stem cells target gastric cancer and deliver epirubicin via tunneling nanotubes for enhanced chemotherapy Curr Stem Cell Res Ther 2024 19 10 1402 13 38173205 10.2174/011574888X287102240101060146 Zhou Y, Li Y, Wang H, Sun H, Su J, Fan Y, et al. Mesenchymal stem cells target gastric cancer and deliver epirubicin via tunneling nanotubes for enhanced chemotherapy. Curr Stem Cell Res Ther. 2024;19(10):1402–13. 38173205 10.2174/011574888X287102240101060146

136. Kalimuthu S Zhu L Oh JM Gangadaran P Lee HW Baek SH Migration of mesenchymal stem cells to tumor xenograft models and in vitro drug delivery by doxorubicin Int J Med Sci 2018 15 10 1051 61 30013447 10.7150/ijms.25760 PMC6036160 Kalimuthu S, Zhu L, Oh JM, Gangadaran P, Lee HW, Baek SH, et al. Migration of mesenchymal stem cells to tumor xenograft models and in vitro drug delivery by doxorubicin. Int J Med Sci. 2018;15(10):1051–61. 30013447 10.7150/ijms.25760 PMC6036160

137. Gil-Chinchilla JI Zapata AG Moraleda JM García-Bernal D Bioengineered mesenchymal stem/stromal cells in anti-cancer therapy: current trends and future prospects Biomolecules 2024 14 7 734 39062449 10.3390/biom14070734 PMC11275142 Gil-Chinchilla JI, Zapata AG, Moraleda JM, García-Bernal D. Bioengineered mesenchymal stem/stromal cells in anti-cancer therapy: current trends and future prospects. Biomolecules. 2024;14(7):734. 39062449 10.3390/biom14070734 PMC11275142

138. Liu W Wang X Feng X Yu J Liu X Jia X Oncolytic adenovirus-mediated intratumoral expression of TRAIL and CD40L enhances immunotherapy by modulating the tumor microenvironment in immunocompetent mouse models Cancer Lett 2022 535 215661 35325845 10.1016/j.canlet.2022.215661 Liu W, Wang X, Feng X, Yu J, Liu X, Jia X, et al. Oncolytic adenovirus-mediated intratumoral expression of TRAIL and CD40L enhances immunotherapy by modulating the tumor microenvironment in immunocompetent mouse models. Cancer Lett. 2022;535:215661. 35325845 10.1016/j.canlet.2022.215661

139. Oraee-Yazdani S Tavanaei R Rostami F Hajarizadeh A Mehrabadi M Akhlaghpasand M Suicide gene therapy using allogeneic adipose tissue-derived mesenchymal stem cell gene delivery vehicles in recurrent glioblastoma multiforme: a first-in-human, dose-escalation, phase I clinical trial J Transl Med 2023 21 1 350 37245011 10.1186/s12967-023-04213-4 PMC10223860 Oraee-Yazdani S, Tavanaei R, Rostami F, Hajarizadeh A, Mehrabadi M, Akhlaghpasand M, et al. Suicide gene therapy using allogeneic adipose tissue-derived mesenchymal stem cell gene delivery vehicles in recurrent glioblastoma multiforme: a first-in-human, dose-escalation, phase I clinical trial. J Transl Med. 2023;21(1):350. 37245011 10.1186/s12967-023-04213-4 PMC10223860

140. Wu Z Huang S Li S Cai J Huang L Wu W Bone marrow mesenchymal stem cell and mononuclear cell combination therapy in patients with type 2 diabetes mellitus: a randomized controlled study with 8-year follow-up Stem Cell Res Ther 2024 15 1 339 39350270 10.1186/s13287-024-03907-w PMC11443831 Wu Z, Huang S, Li S, Cai J, Huang L, Wu W, et al. Bone marrow mesenchymal stem cell and mononuclear cell combination therapy in patients with type 2 diabetes mellitus: a randomized controlled study with 8-year follow-up. Stem Cell Res Ther. 2024;15(1):339. 39350270 10.1186/s13287-024-03907-w PMC11443831

141. Perico N Remuzzi G Griffin MD Cockwell P Maxwell AP Casiraghi F Safety and preliminary efficacy of mesenchymal stromal cell (ORBCEL-M) therapy in diabetic kidney disease: a randomized clinical trial (NEPHSTROM) J Am Soc Nephrol 2023 34 10 1733 51 37560967 10.1681/ASN.0000000000000189 PMC10561817 Perico N, Remuzzi G, Griffin MD, Cockwell P, Maxwell AP, Casiraghi F, et al. Safety and preliminary efficacy of mesenchymal stromal cell (ORBCEL-M) therapy in diabetic kidney disease: a randomized clinical trial (NEPHSTROM). J Am Soc Nephrol. 2023;34(10):1733–51. 37560967 10.1681/ASN.0000000000000189 PMC10561817

142. Arango-Rodríguez ML Mateus LC Sossa CL Becerra-Bayona SM Solarte-David VA Ochoa Vera ME A novel therapeutic management for diabetes patients with chronic limb-threatening ischemia: comparison of autologous bone marrow mononuclear cells versus allogenic wharton jelly-derived mesenchymal stem cells Stem Cell Res Ther 2023 14 1 221 37626416 10.1186/s13287-023-03427-z PMC10464344 Arango-Rodríguez ML, Mateus LC, Sossa CL, Becerra-Bayona SM, Solarte-David VA, Ochoa Vera ME, et al. A novel therapeutic management for diabetes patients with chronic limb-threatening ischemia: comparison of autologous bone marrow mononuclear cells versus allogenic wharton jelly-derived mesenchymal stem cells. Stem Cell Res Ther. 2023;14(1):221. 37626416 10.1186/s13287-023-03427-z PMC10464344

143. Yuan M-Q Song L Wang Z-R Zhang Z-Y Shi M He J Long-term outcomes of mesenchymal stem cell therapy in severe COVID-19 patients: 3-year follow-up of a randomized, double-blind, placebo-controlled trial Stem Cell Res Ther 2025 16 1 94 40001244 10.1186/s13287-025-04148-1 PMC11863646 Yuan M-Q, Song L, Wang Z-R, Zhang Z-Y, Shi M, He J, et al. Long-term outcomes of mesenchymal stem cell therapy in severe COVID-19 patients: 3-year follow-up of a randomized, double-blind, placebo-controlled trial. Stem Cell Res Ther. 2025;16(1):94. 40001244 10.1186/s13287-025-04148-1 PMC11863646

144. Bowdish ME Barkauskas CE Overbey JR Gottlieb RL Osman K Duggal A A randomized trial of mesenchymal stromal cells for moderate to severe acute respiratory distress syndrome from COVID-19 Am J Respir Crit Care Med 2023 207 3 261 70 36099435 10.1164/rccm.202201-0157OC PMC9896641 Bowdish ME, Barkauskas CE, Overbey JR, Gottlieb RL, Osman K, Duggal A, et al. A randomized trial of mesenchymal stromal cells for moderate to severe acute respiratory distress syndrome from COVID-19. Am J Respir Crit Care Med. 2023;207(3):261–70. 36099435 10.1164/rccm.202201-0157OC PMC9896641

145. Vivarelli M Colucci M Algeri M Zotta F Emma F L’Erario I A phase I study of autologous mesenchymal stromal cells for severe steroid-dependent nephrotic syndrome JCI Insight 2023 8 18 e169424 10.1172/jci.insight.169424 37561590 PMC10561718 Vivarelli M, Colucci M, Algeri M, Zotta F, Emma F, L’Erario I, et al. A phase I study of autologous mesenchymal stromal cells for severe steroid-dependent nephrotic syndrome. JCI Insight. 2023;8(18):e169424. 10.1172/jci.insight.169424 37561590 10.1172/jci.insight.169424 PMC10561718

146. Pers Y-M Soler-Rich R Vadalà G Ferreira R Duflos C Picot M-C Allogenic bone marrow-derived mesenchymal stromal cell-based therapy for patients with chronic low back pain: a prospective, multicentre, randomised placebo controlled trial (RESPINE study) Ann Rheum Dis 2024 83 11 1572 83 39393844 10.1136/ard-2024-225771 PMC11503111 Pers Y-M, Soler-Rich R, Vadalà G, Ferreira R, Duflos C, Picot M-C, et al. Allogenic bone marrow-derived mesenchymal stromal cell-based therapy for patients with chronic low back pain: a prospective, multicentre, randomised placebo controlled trial (RESPINE study). Ann Rheum Dis. 2024;83(11):1572–83. 39393844 10.1136/ard-2024-225771 PMC11503111

147. Fenger Carlander A-L Jakobsen KK Todsen T Paaske N Østergaard Madsen AK Bendtsen SK Long-term effectiveness and safety of mesenchymal stromal cell therapy for radiation-induced hyposalivation in head and neck cancer survivors: a randomized phase II trial Clin Cancer Res 2025 31 5 824 31 39751638 10.1158/1078-0432.CCR-24-2663 Fenger Carlander A-L, Jakobsen KK, Todsen T, Paaske N, Østergaard Madsen AK, Bendtsen SK, et al. Long-term effectiveness and safety of mesenchymal stromal cell therapy for radiation-induced hyposalivation in head and neck cancer survivors: a randomized phase II trial. Clin Cancer Res. 2025;31(5):824–31. 39751638 10.1158/1078-0432.CCR-24-2663

148. Attia N Mashal M Puras G Pedraz JL Mesenchymal stem cells as a gene delivery tool: promise, problems, and prospects Pharmaceutics 2021 13 6 843 10.3390/pharmaceutics13060843 34200425 PMC8229096 Attia N, Mashal M, Puras G, Pedraz JL. Mesenchymal stem cells as a gene delivery tool: promise, problems, and prospects. Pharmaceutics. 2021;13(6):843. 10.3390/pharmaceutics13060843 34200425 10.3390/pharmaceutics13060843 PMC8229096

149. Qian L Zhang Z Zhang R Zheng X Xiao B Zhang X Activated STING-containing R-EVs from iPSC -derived MSCs promote antitumor immunity Cancer Lett 2024 597 217081 38909776 10.1016/j.canlet.2024.217081 Qian L, Zhang Z, Zhang R, Zheng X, Xiao B, Zhang X, et al. Activated STING-containing R-EVs from iPSC -derived MSCs promote antitumor immunity. Cancer Lett. 2024;597:217081. 38909776 10.1016/j.canlet.2024.217081

150. Levy O Kuai R Siren EMJ Bhere D Milton Y Nissar N Shattering barriers toward clinically meaningful MSC therapies Sci Adv 2020 6 30 eaba6884 32832666 10.1126/sciadv.aba6884 PMC7439491 Levy O, Kuai R, Siren EMJ, Bhere D, Milton Y, Nissar N, et al. Shattering barriers toward clinically meaningful MSC therapies. Sci Adv. 2020;6(30):eaba6884. 32832666 10.1126/sciadv.aba6884 PMC7439491

151. Clubb JD Gao TA Chen YY Synthetic biology in the engineering of CAR-T and CAR-NK cell therapies: facts and hopes Clin Cancer Res 2023 29 8 1390 402 36454122 10.1158/1078-0432.CCR-22-1491 PMC10106357 Clubb JD, Gao TA, Chen YY. Synthetic biology in the engineering of CAR-T and CAR-NK cell therapies: facts and hopes. Clin Cancer Res. 2023;29(8):1390–402. 36454122 10.1158/1078-0432.CCR-22-1491 PMC10106357

152. Li K Chen C Gao R Yu X Huang Y Chen Z Inhibition of BCL11B induces downregulation of PTK7 and results in growth retardation and apoptosis in T-cell acute lymphoblastic leukemia Biomark Res 2021 9 1 17 33663588 10.1186/s40364-021-00270-3 PMC7934371 Li K, Chen C, Gao R, Yu X, Huang Y, Chen Z, et al. Inhibition of BCL11B induces downregulation of PTK7 and results in growth retardation and apoptosis in T-cell acute lymphoblastic leukemia. Biomark Res. 2021;9(1):17. 33663588 10.1186/s40364-021-00270-3 PMC7934371

153. Philippova J Shevchenko J Sennikov S GD2-targeting therapy: a comparative analysis of approaches and promising directions Front Immunol 2024 15 1371345 38558810 10.3389/fimmu.2024.1371345 PMC10979305 Philippova J, Shevchenko J, Sennikov S. GD2-targeting therapy: a comparative analysis of approaches and promising directions. Front Immunol. 2024;15:1371345. 38558810 10.3389/fimmu.2024.1371345 PMC10979305

154. Aliperta R Cartellieri M Feldmann A Arndt C Koristka S Michalk I Bispecific antibody releasing-mesenchymal stromal cell machinery for retargeting T cells towards acute myeloid leukemia blasts Blood Cancer J 2015 5 9 e348 26383821 10.1038/bcj.2015.73 PMC4648523 Aliperta R, Cartellieri M, Feldmann A, Arndt C, Koristka S, Michalk I, et al. Bispecific antibody releasing-mesenchymal stromal cell machinery for retargeting T cells towards acute myeloid leukemia blasts. Blood Cancer J. 2015;5(9):e348. 26383821 10.1038/bcj.2015.73 PMC4648523

155. Tang S Zhang Y Wang P Tang Q Liu Y Lu F NKG2D-CAR-targeted iPSC-derived MSCs efficiently target solid tumors expressing NKG2D ligand iScience 2025 28 5 112343 40276759 10.1016/j.isci.2025.112343 PMC12020857 Tang S, Zhang Y, Wang P, Tang Q, Liu Y, Lu F, et al. NKG2D-CAR-targeted iPSC-derived MSCs efficiently target solid tumors expressing NKG2D ligand. iScience. 2025;28(5):112343. 40276759 10.1016/j.isci.2025.112343 PMC12020857

156. Niu Z Wu J Zhao Q Zhang J Zhang P Yang Y CAR-based immunotherapy for breast cancer: peculiarities, ongoing investigations, and future strategies Front Immunol 2024 15 1385571 38680498 10.3389/fimmu.2024.1385571 PMC11045891 Niu Z, Wu J, Zhao Q, Zhang J, Zhang P, Yang Y. CAR-based immunotherapy for breast cancer: peculiarities, ongoing investigations, and future strategies. Front Immunol. 2024;15:1385571. 38680498 10.3389/fimmu.2024.1385571 PMC11045891

157. Nawaz W Huang B Xu S Li Y Zhu L Yiqiao H AAV-mediated in vivo CAR gene therapy for targeting human T-cell leukemia Blood Cancer J 2021 11 6 119 34162832 10.1038/s41408-021-00508-1 PMC8222347 Nawaz W, Huang B, Xu S, Li Y, Zhu L, Yiqiao H, et al. AAV-mediated in vivo CAR gene therapy for targeting human T-cell leukemia. Blood Cancer J. 2021;11(6):119. 34162832 10.1038/s41408-021-00508-1 PMC8222347

158. Dong W Kantor B Lentiviral vectors for delivery of gene-editing systems based on CRISPR/Cas: current state and perspectives Viruses 2021 13 7 1288 10.3390/v13071288 34372494 PMC8310029 Dong W, Kantor B. Lentiviral vectors for delivery of gene-editing systems based on CRISPR/Cas: current state and perspectives. Viruses. 2021;13(7):1288. 10.3390/v13071288 34372494 10.3390/v13071288 PMC8310029

159. Ye L Lam SZ Yang L Suzuki K Zou Y Lin Q AAV-mediated delivery of a sleeping beauty transposon and an mRNA-encoded transposase for the engineering of therapeutic immune cells Nat Biomed Eng 2024 8 2 132 48 37430157 10.1038/s41551-023-01058-6 PMC11320892 Ye L, Lam SZ, Yang L, Suzuki K, Zou Y, Lin Q, et al. AAV-mediated delivery of a sleeping beauty transposon and an mRNA-encoded transposase for the engineering of therapeutic immune cells. Nat Biomed Eng. 2024;8(2):132–48. 37430157 10.1038/s41551-023-01058-6 PMC11320892

160. Lei T Wang Y Zhang Y Yang Y Cao J Huang J Leveraging CRISPR gene editing technology to optimize the efficacy, safety and accessibility of CAR T-cell therapy Leukemia 2024 38 12 2517 43 39455854 10.1038/s41375-024-02444-y PMC11588664 Lei T, Wang Y, Zhang Y, Yang Y, Cao J, Huang J, et al. Leveraging CRISPR gene editing technology to optimize the efficacy, safety and accessibility of CAR T-cell therapy. Leukemia. 2024;38(12):2517–43. 39455854 10.1038/s41375-024-02444-y PMC11588664

161. Huang R Wang X Yan H Tan X Ma Y Wang M Safety and efficacy of CD33-targeted CAR-NK cell therapy for relapsed/refractory AML: preclinical evaluation and phase I trial Exp Hematol Oncol 2025 14 1 1 39748428 10.1186/s40164-024-00592-6 PMC11694373 Huang R, Wang X, Yan H, Tan X, Ma Y, Wang M, et al. Safety and efficacy of CD33-targeted CAR-NK cell therapy for relapsed/refractory AML: preclinical evaluation and phase I trial. Exp Hematol Oncol. 2025;14(1):1. 39748428 10.1186/s40164-024-00592-6 PMC11694373

162. Dwyer RM Potter-Beirne SM Harrington KA Lowery AJ Hennessy E Murphy JM Monocyte chemotactic protein-1 secreted by primary breast tumors stimulates migration of mesenchymal stem cells Clin Cancer Res 2007 13 17 5020 7 17785552 10.1158/1078-0432.CCR-07-0731 Dwyer RM, Potter-Beirne SM, Harrington KA, Lowery AJ, Hennessy E, Murphy JM, et al. Monocyte chemotactic protein-1 secreted by primary breast tumors stimulates migration of mesenchymal stem cells. Clin Cancer Res. 2007;13(17):5020–7. 17785552 10.1158/1078-0432.CCR-07-0731

163. Ball SG Shuttleworth CA Kielty CM Vascular endothelial growth factor can signal through platelet-derived growth factor receptors J Cell Biol 2007 177 3 489 500 17470632 10.1083/jcb.200608093 PMC2064818 Ball SG, Shuttleworth CA, Kielty CM. Vascular endothelial growth factor can signal through platelet-derived growth factor receptors. J Cell Biol. 2007;177(3):489–500. 17470632 10.1083/jcb.200608093 PMC2064818

164. Spaeth E Klopp A Dembinski J Andreeff M Marini F Inflammation and tumor microenvironments: defining the migratory itinerary of mesenchymal stem cells Gene Ther 2008 15 10 730 8 18401438 10.1038/gt.2008.39 Spaeth E, Klopp A, Dembinski J, Andreeff M, Marini F. Inflammation and tumor microenvironments: defining the migratory itinerary of mesenchymal stem cells. Gene Ther. 2008;15(10):730–8. 18401438 10.1038/gt.2008.39

165. Wynn RF Hart CA Corradi-Perini C O’Neill L Evans CA Wraith JE A small proportion of mesenchymal stem cells strongly expresses functionally active CXCR4 receptor capable of promoting migration to bone marrow Blood 2004 104 9 2643 5 15251986 10.1182/blood-2004-02-0526 Wynn RF, Hart CA, Corradi-Perini C, O’Neill L, Evans CA, Wraith JE, et al. A small proportion of mesenchymal stem cells strongly expresses functionally active CXCR4 receptor capable of promoting migration to bone marrow. Blood. 2004;104(9):2643–5. 15251986 10.1182/blood-2004-02-0526

166. Rombouts WJC Ploemacher RE Primary murine MSC show highly efficient homing to the bone marrow but lose homing ability following culture Leukemia 2003 17 1 160 70 12529674 10.1038/sj.leu.2402763 Rombouts WJC, Ploemacher RE. Primary murine MSC show highly efficient homing to the bone marrow but lose homing ability following culture. Leukemia. 2003;17(1):160–70. 12529674 10.1038/sj.leu.2402763

167. Xinaris C Morigi M Benedetti V Imberti B Fabricio AS Squarcina E A novel strategy to enhance mesenchymal stem cell migration capacity and promote tissue repair in an injury specific fashion Cell Transplant 2013 22 3 423 36 22889699 10.3727/096368912X653246 Xinaris C, Morigi M, Benedetti V, Imberti B, Fabricio AS, Squarcina E, et al. A novel strategy to enhance mesenchymal stem cell migration capacity and promote tissue repair in an injury specific fashion. Cell Transplant. 2013;22(3):423–36. 22889699 10.3727/096368912X653246

168. Chen W Li M Cheng H Yan Z Cao J Pan B Overexpression of the mesenchymal stem cell Cxcr4 gene in irradiated mice increases the homing capacity of these cells Cell Biochem Biophys 2013 67 3 1181 91 23712865 10.1007/s12013-013-9632-6 Chen W, Li M, Cheng H, Yan Z, Cao J, Pan B, et al. Overexpression of the mesenchymal stem cell Cxcr4 gene in irradiated mice increases the homing capacity of these cells. Cell Biochem Biophys. 2013;67(3):1181–91. 23712865 10.1007/s12013-013-9632-6

169. Zheng X-B He X-W Zhang L-J Qin H-B Lin X-T Liu X-H Bone marrow-derived CXCR4-overexpressing MSCs display increased homing to intestine and ameliorate colitis-associated tumorigenesis in mice Gastroenterol Rep 2019 7 2 127 38 10.1093/gastro/goy017 PMC6454852 30976426 Zheng X-B, He X-W, Zhang L-J, Qin H-B, Lin X-T, Liu X-H, et al. Bone marrow-derived CXCR4-overexpressing MSCs display increased homing to intestine and ameliorate colitis-associated tumorigenesis in mice. Gastroenterol Rep. 2019;7(2):127–38. 10.1093/gastro/goy017 PMC6454852 30976426

170. Najar M Raicevic G Fayyad-Kazan H Bron D Toungouz M Lagneaux L Mesenchymal stromal cells and immunomodulation: a gathering of regulatory immune cells Cytotherapy 2016 18 2 160 71 26794710 10.1016/j.jcyt.2015.10.011 Najar M, Raicevic G, Fayyad-Kazan H, Bron D, Toungouz M, Lagneaux L. Mesenchymal stromal cells and immunomodulation: a gathering of regulatory immune cells. Cytotherapy. 2016;18(2):160–71. 26794710 10.1016/j.jcyt.2015.10.011

171. Ma X Chen J Liu J Xu B Liang X Yang X IL-8/CXCR2 mediates tropism of human bone marrow-derived mesenchymal stem cells toward CD133+ /CD44 + colon cancer stem cells J Cell Physiol 2021 236 4 3114 28 33078417 10.1002/jcp.30080 Ma X, Chen J, Liu J, Xu B, Liang X, Yang X, et al. IL-8/CXCR2 mediates tropism of human bone marrow-derived mesenchymal stem cells toward CD133+ /CD44 + colon cancer stem cells. J Cell Physiol. 2021;236(4):3114–28. 33078417 10.1002/jcp.30080

172. Chen M Liu X Peng N Zhang T Mou J He H Construction of CD19 targeted dual- and enhanced dual-antibodies and their efficiency in the treatment of B cell malignancy Exp Hematol Oncol 2023 12 1 64 37488603 10.1186/s40164-023-00423-0 PMC10367426 Chen M, Liu X, Peng N, Zhang T, Mou J, He H, et al. Construction of CD19 targeted dual- and enhanced dual-antibodies and their efficiency in the treatment of B cell malignancy. Exp Hematol Oncol. 2023;12(1):64. 37488603 10.1186/s40164-023-00423-0 PMC10367426

173. Hegde M Mukherjee M Grada Z Pignata A Landi D Navai SA Tandem CAR t cells targeting HER2 and IL13Rα2 mitigate tumor antigen escape J Clin Invest 2019 129 8 3464 31264975 10.1172/JCI131246 PMC6668664 Hegde M, Mukherjee M, Grada Z, Pignata A, Landi D, Navai SA, et al. Tandem CAR t cells targeting HER2 and IL13Rα2 mitigate tumor antigen escape. J Clin Invest. 2019;129(8):3464. 31264975 10.1172/JCI131246 PMC6668664

174. Tousley AM Rotiroti MC Labanieh L Rysavy LW Kim W-J Lareau C Co-opting signalling molecules enables logic-gated control of CAR T cells Nature 2023 615 7952 507 16 36890224 10.1038/s41586-023-05778-2 PMC10564584 Tousley AM, Rotiroti MC, Labanieh L, Rysavy LW, Kim W-J, Lareau C, et al. Co-opting signalling molecules enables logic-gated control of CAR T cells. Nature. 2023;615(7952):507–16. 36890224 10.1038/s41586-023-05778-2 PMC10564584

175. Bangayan NJ Wang L Burton Sojo G Noguchi M Cheng D Ta L Dual-inhibitory domain iCARs improve the efficiency of the and-not gate CAR T strategy Proc Natl Acad Sci U S A 2023 120 47 e2312374120 37963244 10.1073/pnas.2312374120 PMC10666036 Bangayan NJ, Wang L, Burton Sojo G, Noguchi M, Cheng D, Ta L, et al. Dual-inhibitory domain iCARs improve the efficiency of the and-not gate CAR T strategy. Proc Natl Acad Sci U S A. 2023;120(47):e2312374120. 37963244 10.1073/pnas.2312374120 PMC10666036

176. Wang L Zhang L Dunmall LC Wang YY Fan Z Cheng Z The dilemmas and possible solutions for CAR-T cell therapy application in solid tumors Cancer Lett 2024 591 216871 38604310 10.1016/j.canlet.2024.216871 Wang L, Zhang L, Dunmall LC, Wang YY, Fan Z, Cheng Z, et al. The dilemmas and possible solutions for CAR-T cell therapy application in solid tumors. Cancer Lett. 2024;591:216871. 38604310 10.1016/j.canlet.2024.216871

177. Roybal KT Williams JZ Morsut L Rupp LJ Kolinko I Choe JH Engineering T cells with customized therapeutic response programs using synthetic Notch receptors Cell 2016 167 2 419 432.e16 10.1016/j.cell.2016.09.011 27693353 PMC5072533 Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH, et al. Engineering T cells with customized therapeutic response programs using synthetic Notch receptors. Cell. 2016 Oct 6;167(2):419–432.e16. 10.1016/j.cell.2016.09.011 27693353 10.1016/j.cell.2016.09.011 PMC5072533

178. Li X Li W Xu L Song Y Chimeric antigen receptor-immune cells against solid tumors: structures, mechanisms, recent advances, and future developments Chin Med J (Engl) 2024 137 11 1285 302 37640679 10.1097/CM9.0000000000002818 PMC11191032 Li X, Li W, Xu L, Song Y. Chimeric antigen receptor-immune cells against solid tumors: structures, mechanisms, recent advances, and future developments. Chin Med J (Engl). 2024;137(11):1285–302. 37640679 10.1097/CM9.0000000000002818 PMC11191032

179. Keshavarz A Salehi A Khosravi S Shariati Y Nasrabadi N Kahrizi MS Recent findings on chimeric antigen receptor (CAR)-engineered immune cell therapy in solid tumors and hematological malignancies Stem Cell Res Ther 2022 13 1 482 36153626 10.1186/s13287-022-03163-w PMC9509604 Keshavarz A, Salehi A, Khosravi S, Shariati Y, Nasrabadi N, Kahrizi MS, et al. Recent findings on chimeric antigen receptor (CAR)-engineered immune cell therapy in solid tumors and hematological malignancies. Stem Cell Res Ther. 2022;13(1):482. 36153626 10.1186/s13287-022-03163-w PMC9509604

180. Han X Bryson PD Zhao Y Cinay GE Li S Guo Y Masked chimeric antigen receptor for tumor-specific activation Mol Ther 2017 25 1 274 84 28129121 10.1016/j.ymthe.2016.10.011 PMC5363190 Han X, Bryson PD, Zhao Y, Cinay GE, Li S, Guo Y, et al. Masked chimeric antigen receptor for tumor-specific activation. Mol Ther. 2017;25(1):274–84. 28129121 10.1016/j.ymthe.2016.10.011 PMC5363190

181. Volta L Myburgh R Pellegrino C Koch C Maurer M Manfredi F Efficient combinatorial adaptor-mediated targeting of acute myeloid leukemia with CAR T-cells Leukemia 2024 38 12 2598 613 39294295 10.1038/s41375-024-02409-1 PMC11588662 Volta L, Myburgh R, Pellegrino C, Koch C, Maurer M, Manfredi F, et al. Efficient combinatorial adaptor-mediated targeting of acute myeloid leukemia with CAR T-cells. Leukemia. 2024;38(12):2598–613. 39294295 10.1038/s41375-024-02409-1 PMC11588662

182. Garcia J Daniels J Lee Y Zhu I Cheng K Liu Q Naturally occurring T cell mutations enhance engineered T cell therapies Nature 2024 626 7999 626 34 38326614 10.1038/s41586-024-07018-7 PMC11573425 Garcia J, Daniels J, Lee Y, Zhu I, Cheng K, Liu Q, et al. Naturally occurring T cell mutations enhance engineered T cell therapies. Nature. 2024;626(7999):626–34. 38326614 10.1038/s41586-024-07018-7 PMC11573425

183. Lai P Chen X Guo L Wang Y Liu X Liu Y A potent immunomodulatory role of exosomes derived from mesenchymal stromal cells in preventing cGVHD J Hematol Oncol 2018 11 1 135 30526632 10.1186/s13045-018-0680-7 PMC6286548 Lai P, Chen X, Guo L, Wang Y, Liu X, Liu Y, et al. A potent immunomodulatory role of exosomes derived from mesenchymal stromal cells in preventing cGVHD. J Hematol Oncol. 2018;11(1):135. 30526632 10.1186/s13045-018-0680-7 PMC6286548

184. Lyu T Wang Y Li D Yang H Qin B Zhang W Exosomes from BM-MSCs promote acute myeloid leukemia cell proliferation, invasion and chemoresistance via upregulation of S100A4 Exp Hematol Oncol 2021 10 1 24 33789743 10.1186/s40164-021-00220-7 PMC8011411 Lyu T, Wang Y, Li D, Yang H, Qin B, Zhang W, et al. Exosomes from BM-MSCs promote acute myeloid leukemia cell proliferation, invasion and chemoresistance via upregulation of S100A4. Exp Hematol Oncol. 2021;10(1):24. 33789743 10.1186/s40164-021-00220-7 PMC8011411

185. Yin K Wang S Zhao RC Exosomes from mesenchymal stem/stromal cells: a new therapeutic paradigm Biomark Res 2019 7 8 30992990 10.1186/s40364-019-0159-x PMC6450000 Yin K, Wang S, Zhao RC. Exosomes from mesenchymal stem/stromal cells: a new therapeutic paradigm. Biomark Res. 2019;7:8. 30992990 10.1186/s40364-019-0159-x PMC6450000

186. Sajjad U Ahmed M Iqbal MZ Riaz M Mustafa M Biedermann T Exploring mesenchymal stem cells homing mechanisms and improvement strategies Stem Cells Transl Med 2024 13 12 1161 77 39550211 10.1093/stcltm/szae045 PMC11631218 Sajjad U, Ahmed M, Iqbal MZ, Riaz M, Mustafa M, Biedermann T, et al. Exploring mesenchymal stem cells homing mechanisms and improvement strategies. Stem Cells Transl Med. 2024;13(12):1161–77. 39550211 10.1093/stcltm/szae045 PMC11631218

187. Chen J Qiu S Li W Wang K Zhang Y Yang H Tuning charge density of chimeric antigen receptor optimizes tonic signaling and CAR-T cell fitness Cell Res 2023 33 5 341 54 36882513 10.1038/s41422-023-00789-0 PMC10156745 Chen J, Qiu S, Li W, Wang K, Zhang Y, Yang H, et al. Tuning charge density of chimeric antigen receptor optimizes tonic signaling and CAR-T cell fitness. Cell Res. 2023;33(5):341–54. 36882513 10.1038/s41422-023-00789-0 PMC10156745

188. Li W Qiu S Chen J Jiang S Chen W Jiang J Chimeric antigen receptor designed to prevent ubiquitination and downregulation showed durable antitumor efficacy Immunity 2020 53 2 456 470.e6 10.1016/j.immuni.2020.07.011 32758419 Li W, Qiu S, Chen J, Jiang S, Chen W, Jiang J, et al. Chimeric antigen receptor designed to prevent ubiquitination and downregulation showed durable antitumor efficacy. Immunity. 2020;53(2):456–470.e6. 10.1016/j.immuni.2020.07.011 32758419 10.1016/j.immuni.2020.07.011

189. Dai Y Liu Y An L Zhong F Zhang X Lou S Afatinib boosts CAR-T cell antitumor therapeutic efficacy via metabolism and fate reprogramming J Immunother Cancer 2024 12 11 e009949 10.1136/jitc-2024-009949 39551605 PMC11574435 Dai Y, Liu Y, An L, Zhong F, Zhang X, Lou S. Afatinib boosts CAR-T cell antitumor therapeutic efficacy via metabolism and fate reprogramming. J Immunother Cancer. 2024;12(11):e009949. 10.1136/jitc-2024-009949 39551605 10.1136/jitc-2024-009949 PMC11574435

190. Liu Y An L Wang X Dai Y Zhang C Wen Q Engineering a controllable and reversible switch for CAR-based cellular immunotherapies via a genetic code expansion system J Hematol Oncol 2024 17 1 122 39696585 10.1186/s13045-024-01648-0 PMC11657976 Liu Y, An L, Wang X, Dai Y, Zhang C, Wen Q, et al. Engineering a controllable and reversible switch for CAR-based cellular immunotherapies via a genetic code expansion system. J Hematol Oncol. 2024;17(1):122. 39696585 10.1186/s13045-024-01648-0 PMC11657976

191. Labanieh L Mackall CL CAR immune cells: design principles, resistance and the next generation Nature 2023 614 7949 635 48 36813894 10.1038/s41586-023-05707-3 Labanieh L, Mackall CL. CAR immune cells: design principles, resistance and the next generation. Nature. 2023;614(7949):635–48. 36813894 10.1038/s41586-023-05707-3

192. Steffin D Ghatwai N Montalbano A Rathi P Courtney AN Arnett AB Interleukin-15-armoured GPC3 CAR T cells for patients with solid cancers Nature 2024 637 8047 940 946 10.1038/s41586-024-08261-8 39604730 PMC12704925 Steffin D, Ghatwai N, Montalbano A, Rathi P, Courtney AN, Arnett AB, et al. Interleukin-15-armoured GPC3 CAR T cells for patients with solid cancers. Nature. 2025;637(8047):940–946. 10.1038/s41586-024-08261-8 39604730 10.1038/s41586-024-08261-8 PMC12704925

193. Doan AE Mueller KP Chen AY Rouin GT Chen Y Daniel B FOXO1 is a master regulator of memory programming in CAR T cells Nature 2024 629 8010 211 8 38600391 10.1038/s41586-024-07300-8 PMC11062920 Doan AE, Mueller KP, Chen AY, Rouin GT, Chen Y, Daniel B, et al. FOXO1 is a master regulator of memory programming in CAR T cells. Nature. 2024;629(8010):211–8. 38600391 10.1038/s41586-024-07300-8 PMC11062920

194. Chan JD Scheffler CM Munoz I Sek K Lee JN Huang Y-K Foxo1 enhances CAR T cell stemness, metabolic fitness and efficacy Nature 2024 629 8010 201 10 38600376 10.1038/s41586-024-07242-1 PMC11062918 Chan JD, Scheffler CM, Munoz I, Sek K, Lee JN, Huang Y-K, et al. Foxo1 enhances CAR T cell stemness, metabolic fitness and efficacy. Nature. 2024;629(8010):201–10. 38600376 10.1038/s41586-024-07242-1 PMC11062918

195. Ahn T Bae E-A Seo H Decoding and overcoming T cell exhaustion: epigenetic and transcriptional dynamics in CAR-T cells against solid tumors Mol Ther 2024 32 6 1617 27 38582965 10.1016/j.ymthe.2024.04.004 PMC11184340 Ahn T, Bae E-A, Seo H. Decoding and overcoming T cell exhaustion: epigenetic and transcriptional dynamics in CAR-T cells against solid tumors. Mol Ther. 2024;32(6):1617–27. 38582965 10.1016/j.ymthe.2024.04.004 PMC11184340

196. Sui S Zhong M Zhong S Peng X Mao L Chen C BRD4 inhibitor reduces exhaustion and blocks terminal differentiation in CAR-T cells by modulating BATF and EGR1 Biomark Res 2024 12 1 124 39407311 10.1186/s40364-024-00667-w PMC11476310 Sui S, Zhong M, Zhong S, Peng X, Mao L, Chen C, et al. BRD4 inhibitor reduces exhaustion and blocks terminal differentiation in CAR-T cells by modulating BATF and EGR1. Biomark Res. 2024;12(1):124. 39407311 10.1186/s40364-024-00667-w PMC11476310

197. Lee SS Al Halawani A Teo JD Weiss AS Yeo GC The matrix protein tropoelastin prolongs mesenchymal stromal cell vitality and delays senescence during replicative aging Adv Sci (Weinh) 2024 11 39 e2402168 39120048 10.1002/advs.202402168 PMC11497112 Lee SS, Al Halawani A, Teo JD, Weiss AS, Yeo GC. The matrix protein tropoelastin prolongs mesenchymal stromal cell vitality and delays senescence during replicative aging. Adv Sci (Weinh). 2024;11(39):e2402168. 39120048 10.1002/advs.202402168 PMC11497112

198. Jin HJ Kwon JH Kim M Bae YK Choi SJ Oh W Downregulation of melanoma cell adhesion molecule (MCAM/CD146) accelerates cellular senescence in human umbilical cord blood-derived mesenchymal stem cells Stem Cells Transl Med 2016 5 4 427 39 26941359 10.5966/sctm.2015-0109 PMC4798732 Jin HJ, Kwon JH, Kim M, Bae YK, Choi SJ, Oh W, et al. Downregulation of melanoma cell adhesion molecule (MCAM/CD146) accelerates cellular senescence in human umbilical cord blood-derived mesenchymal stem cells. Stem Cells Transl Med. 2016;5(4):427–39. 26941359 10.5966/sctm.2015-0109 PMC4798732

199. Okada M Kim HW Matsu-ura K Wang Y-G Xu M Ashraf M Abrogation of age-induced microrna-195 rejuvenates the senescent mesenchymal stem cells by reactivating telomerase Stem Cells 2016 34 1 148 59 26390028 10.1002/stem.2211 PMC4797648 Okada M, Kim HW, Matsu-ura K, Wang Y-G, Xu M, Ashraf M. Abrogation of age-induced microrna-195 rejuvenates the senescent mesenchymal stem cells by reactivating telomerase. Stem Cells. 2016;34(1):148–59. 26390028 10.1002/stem.2211 PMC4797648

200. Liang X Ding Y Lin F Zhang Y Zhou X Meng Q Overexpression of ERBB4 rejuvenates aged mesenchymal stem cells and enhances angiogenesis via PI3K/AKT and MAPK/ERK pathways FASEB J 2019 33 3 4559 70 30566395 10.1096/fj.201801690R Liang X, Ding Y, Lin F, Zhang Y, Zhou X, Meng Q, et al. Overexpression of ERBB4 rejuvenates aged mesenchymal stem cells and enhances angiogenesis via PI3K/AKT and MAPK/ERK pathways. FASEB J. 2019;33(3):4559–70. 30566395 10.1096/fj.201801690R

201. Fu L Hu Y Song M Liu Z Zhang W Yu F-X Up-regulation of FOXD1 by YAP alleviates senescence and osteoarthritis PLoS Biol 2019 17 4 e3000201 30933975 10.1371/journal.pbio.3000201 PMC6459557 Fu L, Hu Y, Song M, Liu Z, Zhang W, Yu F-X, et al. Up-regulation of FOXD1 by YAP alleviates senescence and osteoarthritis. PLoS Biol. 2019;17(4):e3000201. 30933975 10.1371/journal.pbio.3000201 PMC6459557

202. Samal JRK Rangasami VK Samanta S Varghese OP Oommen OP Discrepancies on the role of oxygen gradient and culture condition on mesenchymal stem cell fate Adv Healthc Mater 2021 10 6 e2002058 33533187 10.1002/adhm.202002058 PMC11469238 Samal JRK, Rangasami VK, Samanta S, Varghese OP, Oommen OP. Discrepancies on the role of oxygen gradient and culture condition on mesenchymal stem cell fate. Adv Healthc Mater. 2021;10(6):e2002058. 33533187 10.1002/adhm.202002058 PMC11469238

203. Gharibi B Hughes FJ Effects of medium supplements on proliferation, differentiation potential, and in vitro expansion of mesenchymal stem cells Stem Cells Transl Med 2012 1 11 771 82 23197689 10.5966/sctm.2010-0031 PMC3659663 Gharibi B, Hughes FJ. Effects of medium supplements on proliferation, differentiation potential, and in vitro expansion of mesenchymal stem cells. Stem Cells Transl Med. 2012;1(11):771–82. 23197689 10.5966/sctm.2010-0031 PMC3659663

204. Yuan H-F Zhai C Yan X-L Zhao D-D Wang J-X Zeng Q SIRT1 is required for long-term growth of human mesenchymal stem cells J Mol Med (Berl) 2012 90 4 389 400 22038097 10.1007/s00109-011-0825-4 Yuan H-F, Zhai C, Yan X-L, Zhao D-D, Wang J-X, Zeng Q, et al. SIRT1 is required for long-term growth of human mesenchymal stem cells. J Mol Med (Berl). 2012;90(4):389–400. 22038097 10.1007/s00109-011-0825-4

205. Denu RA Sirt3 enhances mesenchymal stem cell longevity and differentiation Oxid Med Cell Longev 2017 2017 5841716 28717408 10.1155/2017/5841716 PMC5499245 Denu RA. Sirt3 enhances mesenchymal stem cell longevity and differentiation. Oxid Med Cell Longev. 2017;2017:5841716. 28717408 10.1155/2017/5841716 PMC5499245

206. Yang B Zwaans BMM Eckersdorff M Lombard DB The sirtuin SIRT6 deacetylates H3 K56Ac in vivo to promote genomic stability Cell Cycle 2009 8 16 2662 3 19597350 10.4161/cc.8.16.9329 PMC2728171 Yang B, Zwaans BMM, Eckersdorff M, Lombard DB. The sirtuin SIRT6 deacetylates H3 K56Ac in vivo to promote genomic stability. Cell Cycle. 2009;8(16):2662–3. 19597350 10.4161/cc.8.16.9329 PMC2728171

207. Bi S Liu Z Wu Z Wang Z Liu X Wang S Sirt7 antagonizes human stem cell aging as a heterochromatin stabilizer Protein Cell 2020 11 7 483 504 32504224 10.1007/s13238-020-00728-4 PMC7305295 Bi S, Liu Z, Wu Z, Wang Z, Liu X, Wang S, et al. Sirt7 antagonizes human stem cell aging as a heterochromatin stabilizer. Protein Cell. 2020;11(7):483–504. 32504224 10.1007/s13238-020-00728-4 PMC7305295

208. Calcat-I-Cervera S Rendra E Scaccia E Amadeo F Hanson V Wilm B Harmonised culture procedures minimise but do not eliminate mesenchymal stromal cell donor and tissue variability in a decentralised multicentre manufacturing approach Stem Cell Res Ther 2023 14 1 120 37143116 10.1186/s13287-023-03352-1 PMC10161493 Calcat-I-Cervera S, Rendra E, Scaccia E, Amadeo F, Hanson V, Wilm B, et al. Harmonised culture procedures minimise but do not eliminate mesenchymal stromal cell donor and tissue variability in a decentralised multicentre manufacturing approach. Stem Cell Res Ther. 2023;14(1):120. 37143116 10.1186/s13287-023-03352-1 PMC10161493

209. Česnik AB Švajger U The issue of heterogeneity of MSC-based advanced therapy medicinal products-a review Front Cell Dev Biol 2024 12 1400347 39129786 10.3389/fcell.2024.1400347 PMC11310176 Česnik AB, Švajger U. The issue of heterogeneity of MSC-based advanced therapy medicinal products-a review. Front Cell Dev Biol. 2024;12:1400347. 39129786 10.3389/fcell.2024.1400347 PMC11310176

210. Song I Rim J Lee J Jang I Jung B Kim K Therapeutic potential of human fetal mesenchymal stem cells in musculoskeletal disorders: a narrative review Int J Mol Sci 2022 23 3 1439 10.3390/ijms23031439 35163361 PMC8835918 Song I, Rim J, Lee J, Jang I, Jung B, Kim K, et al. Therapeutic potential of human fetal mesenchymal stem cells in musculoskeletal disorders: a narrative review. Int J Mol Sci. 2022;23(3):1439. 10.3390/ijms23031439 35163361 10.3390/ijms23031439 PMC8835918

211. Giri J Galipeau J Mesenchymal stromal cell therapeutic potency is dependent upon viability, route of delivery, and immune match Blood Adv 2020 4 9 1987 97 32384543 10.1182/bloodadvances.2020001711 PMC7218435 Giri J, Galipeau J. Mesenchymal stromal cell therapeutic potency is dependent upon viability, route of delivery, and immune match. Blood Adv. 2020;4(9):1987–97. 32384543 10.1182/bloodadvances.2020001711 PMC7218435

212. Bahsoun S Coopman K Akam EC Quantitative assessment of the impact of cryopreservation on human bone marrow-derived mesenchymal stem cells: up to 24 h post-thaw and beyond Stem Cell Res Ther 2020 11 1 540 33317625 10.1186/s13287-020-02054-2 PMC7734731 Bahsoun S, Coopman K, Akam EC. Quantitative assessment of the impact of cryopreservation on human bone marrow-derived mesenchymal stem cells: up to 24 h post-thaw and beyond. Stem Cell Res Ther. 2020;11(1):540. 33317625 10.1186/s13287-020-02054-2 PMC7734731

213. Gonzalez-Martinez N Gibson MI Post-thaw application of ROCK-inhibitors increases cryopreserved T-cell yield RSC Med Chem 2023 14 10 2058 67 37859712 10.1039/d3md00378g PMC10583820 Gonzalez-Martinez N, Gibson MI. Post-thaw application of ROCK-inhibitors increases cryopreserved T-cell yield. RSC Med Chem. 2023;14(10):2058–67. 37859712 10.1039/d3md00378g PMC10583820

214. Heng BC Effect of Rho-associated kinase (ROCK) inhibitor Y-27632 on the post-thaw viability of cryopreserved human bone marrow-derived mesenchymal stem cells Tissue Cell 2009 41 5 376 80 19261317 10.1016/j.tice.2009.01.004 Heng BC. Effect of Rho-associated kinase (ROCK) inhibitor Y-27632 on the post-thaw viability of cryopreserved human bone marrow-derived mesenchymal stem cells. Tissue Cell. 2009;41(5):376–80. 19261317 10.1016/j.tice.2009.01.004

215. Cottle C Porter AP Lipat A Turner-Lyles C Nguyen J Moll G Impact of cryopreservation and freeze-thawing on therapeutic properties of mesenchymal stromal/stem cells and other common cellular therapeutics Curr Stem Cell Rep 2022 8 2 72 92 35502223 10.1007/s40778-022-00212-1 PMC9045030 Cottle C, Porter AP, Lipat A, Turner-Lyles C, Nguyen J, Moll G, et al. Impact of cryopreservation and freeze-thawing on therapeutic properties of mesenchymal stromal/stem cells and other common cellular therapeutics. Curr Stem Cell Rep. 2022;8(2):72–92. 35502223 10.1007/s40778-022-00212-1 PMC9045030

216. Marquez-Curtis LA Janowska-Wieczorek A McGann LE Elliott JAW Mesenchymal stromal cells derived from various tissues: biological, clinical and cryopreservation aspects Cryobiology 2015 71 2 181 97 26186998 10.1016/j.cryobiol.2015.07.003 Marquez-Curtis LA, Janowska-Wieczorek A, McGann LE, Elliott JAW. Mesenchymal stromal cells derived from various tissues: biological, clinical and cryopreservation aspects. Cryobiology. 2015;71(2):181–97. 26186998 10.1016/j.cryobiol.2015.07.003

217. Levine BL Miskin J Wonnacott K Keir C Global manufacturing of CAR T cell therapy Mol Ther 2017 4 92 101 10.1016/j.omtm.2016.12.006 PMC5363291 28344995 Levine BL, Miskin J, Wonnacott K, Keir C. Global manufacturing of CAR T cell therapy. Mol Ther Methods Clin Dev. 2016;4:92–101. 10.1016/j.omtm.2016.12.006 10.1016/j.omtm.2016.12.006 PMC5363291 28344995

218. Bexte T Ullrich E Empowering virus-free CAR immune cell therapies Mol Ther 2024 32 6 1609 11 38795701 10.1016/j.ymthe.2024.05.023 PMC11184381 Bexte T, Ullrich E. Empowering virus-free CAR immune cell therapies. Mol Ther. 2024;32(6):1609–11. 38795701 10.1016/j.ymthe.2024.05.023 PMC11184381

219. Enriquez-Rodriguez L Attia N Gallego I Mashal M Maldonado I Puras G Expanding the horizon of transient CAR T therapeutics using virus-free technology Biotechnol Adv 2024 72 108350 38537878 10.1016/j.biotechadv.2024.108350 Enriquez-Rodriguez L, Attia N, Gallego I, Mashal M, Maldonado I, Puras G, et al. Expanding the horizon of transient CAR T therapeutics using virus-free technology. Biotechnol Adv. 2024;72:108350. 38537878 10.1016/j.biotechadv.2024.108350

220. McErlean EM McCarthy HO Non-viral approaches in CAR-NK cell engineering: connecting natural killer cell biology and gene delivery J Nanobiotechnol 2024 22 1 552 10.1186/s12951-024-02746-4 PMC11384716 39256765 McErlean EM, McCarthy HO. Non-viral approaches in CAR-NK cell engineering: connecting natural killer cell biology and gene delivery. J Nanobiotechnol. 2024;22(1):552. 10.1186/s12951-024-02746-4 PMC11384716 39256765

221. Bexte T Botezatu L Miskey C Gierschek F Moter A Wendel P Engineering of potent CAR NK cells using non-viral sleeping beauty transposition from minimalistic DNA vectors Mol Ther 2024 32 7 2357 72 38751112 10.1016/j.ymthe.2024.05.022 PMC11287004 Bexte T, Botezatu L, Miskey C, Gierschek F, Moter A, Wendel P, et al. Engineering of potent CAR NK cells using non-viral sleeping beauty transposition from minimalistic DNA vectors. Mol Ther. 2024;32(7):2357–72. 38751112 10.1016/j.ymthe.2024.05.022 PMC11287004

222. Skeate JG Pomeroy EJ Slipek NJ Jones BJ Wick BJ Chang J-W Evolution of the clinical-stage hyperactive TcBuster transposase as a platform for robust non-viral production of adoptive cellular therapies Mol Ther 2024 32 6 1817 34 38627969 10.1016/j.ymthe.2024.04.024 PMC11184336 Skeate JG, Pomeroy EJ, Slipek NJ, Jones BJ, Wick BJ, Chang J-W, et al. Evolution of the clinical-stage hyperactive TcBuster transposase as a platform for robust non-viral production of adoptive cellular therapies. Mol Ther. 2024;32(6):1817–34. 38627969 10.1016/j.ymthe.2024.04.024 PMC11184336

223. Krueger TEG Thorek DLJ Denmeade SR Isaacs JT Brennen WN Concise review: mesenchymal stem cell-based drug delivery: the good, the bad, the ugly, and the promise Stem Cells Transl Med 2018 7 9 651 63 30070053 10.1002/sctm.18-0024 PMC6127224 Krueger TEG, Thorek DLJ, Denmeade SR, Isaacs JT, Brennen WN. Concise review: mesenchymal stem cell-based drug delivery: the good, the bad, the ugly, and the promise. Stem Cells Transl Med. 2018;7(9):651–63. 30070053 10.1002/sctm.18-0024 PMC6127224

224. Gholamrezanezhad A Mirpour S Bagheri M Mohamadnejad M Alimoghaddam K Abdolahzadeh L In vivo tracking of 111In-oxine labeled mesenchymal stem cells following infusion in patients with advanced cirrhosis Nucl Med Biol 2011 38 7 961 7 21810549 10.1016/j.nucmedbio.2011.03.008 Gholamrezanezhad A, Mirpour S, Bagheri M, Mohamadnejad M, Alimoghaddam K, Abdolahzadeh L, et al. In vivo tracking of 111In-oxine labeled mesenchymal stem cells following infusion in patients with advanced cirrhosis. Nucl Med Biol. 2011;38(7):961–7. 21810549 10.1016/j.nucmedbio.2011.03.008

225. Zhuang W-Z Lin Y-H Su L-J Wu M-S Jeng H-Y Chang H-C Mesenchymal stem/stromal cell-based therapy: mechanism, systemic safety and biodistribution for precision clinical applications J Biomed Sci 2021 28 1 28 33849537 10.1186/s12929-021-00725-7 PMC8043779 Zhuang W-Z, Lin Y-H, Su L-J, Wu M-S, Jeng H-Y, Chang H-C, et al. Mesenchymal stem/stromal cell-based therapy: mechanism, systemic safety and biodistribution for precision clinical applications. J Biomed Sci. 2021;28(1):28. 33849537 10.1186/s12929-021-00725-7 PMC8043779

226. Zanetti A Grata M Etling EB Panday R Villanueva FS Toma C Suspension-expansion of bone marrow results in small mesenchymal stem cells exhibiting increased transpulmonary passage following intravenous administration Tissue Eng Part C Methods 2015 21 7 683 92 25567723 10.1089/ten.tec.2014.0344 PMC4499773 Zanetti A, Grata M, Etling EB, Panday R, Villanueva FS, Toma C. Suspension-expansion of bone marrow results in small mesenchymal stem cells exhibiting increased transpulmonary passage following intravenous administration. Tissue Eng Part C Methods. 2015;21(7):683–92. 25567723 10.1089/ten.tec.2014.0344 PMC4499773

227. Li Z Hu X Mao J Liu X Zhang L Liu J Optimization of mesenchymal stem cells (MSCs) delivery dose and route in mice with acute liver injury by bioluminescence imaging Mol Imaging Biol 2015 17 2 185 94 25273323 10.1007/s11307-014-0792-6 Li Z, Hu X, Mao J, Liu X, Zhang L, Liu J, et al. Optimization of mesenchymal stem cells (MSCs) delivery dose and route in mice with acute liver injury by bioluminescence imaging. Mol Imaging Biol. 2015;17(2):185–94. 25273323 10.1007/s11307-014-0792-6

228. Primavera R Regmi S Yarani R Levitte S Wang J Ganguly A Precision delivery of human bone marrow-derived mesenchymal stem cells into the pancreas via intra-arterial injection prevents the onset of diabetes Stem Cells Transl Med 2024 13 6 559 71 38530131 10.1093/stcltm/szae020 PMC11165159 Primavera R, Regmi S, Yarani R, Levitte S, Wang J, Ganguly A, et al. Precision delivery of human bone marrow-derived mesenchymal stem cells into the pancreas via intra-arterial injection prevents the onset of diabetes. Stem Cells Transl Med. 2024;13(6):559–71. 38530131 10.1093/stcltm/szae020 PMC11165159

229. Wilson T Stark C Holmbom J Rosling A Kuusilehto A Tirri T Fate of bone marrow-derived stromal cells after intraperitoneal infusion or implantation into femoral bone defects in the host animal J Tissue Eng 2010 2010 345806 21350643 10.4061/2010/345806 PMC3042670 Wilson T, Stark C, Holmbom J, Rosling A, Kuusilehto A, Tirri T, et al. Fate of bone marrow-derived stromal cells after intraperitoneal infusion or implantation into femoral bone defects in the host animal. J Tissue Eng. 2010;2010:345806. 21350643 10.4061/2010/345806 PMC3042670

230. Kimura K Kishida T Wakao J Tanaka T Higashi M Fumino S Tumor-homing effect of human mesenchymal stem cells in a TH-MYCN mouse model of neuroblastoma J Pediatr Surg 2016 51 12 2068 73 27686479 10.1016/j.jpedsurg.2016.09.041 Kimura K, Kishida T, Wakao J, Tanaka T, Higashi M, Fumino S, et al. Tumor-homing effect of human mesenchymal stem cells in a TH-MYCN mouse model of neuroblastoma. J Pediatr Surg. 2016;51(12):2068–73. 27686479 10.1016/j.jpedsurg.2016.09.041

231. Wang X Gao J Ouyang X Wang J Sun X Lv Y Mesenchymal stem cells loaded with paclitaxel-poly(lactic-co-glycolic acid) nanoparticles for glioma-targeting therapy Int J Nanomed 2018 13 5231 48 10.2147/IJN.S167142 PMC6136913 30237710 Wang X, Gao J, Ouyang X, Wang J, Sun X, Lv Y. Mesenchymal stem cells loaded with paclitaxel-poly(lactic-co-glycolic acid) nanoparticles for glioma-targeting therapy. Int J Nanomed. 2018;13:5231–48. 10.2147/IJN.S167142 PMC6136913 30237710

232. Dali P Shende P Advances in stem cell therapy for brain diseases via the intranasal route Curr Pharm Biotechnol 2021 22 11 1466 81 33342406 10.2174/1389201021666201218130947 Dali P, Shende P. Advances in stem cell therapy for brain diseases via the intranasal route. Curr Pharm Biotechnol. 2021;22(11):1466–81. 33342406 10.2174/1389201021666201218130947

233. Seo SH Kim KS Park SH Suh YS Kim SJ Jeun SS The effects of mesenchymal stem cells injected via different routes on modified IL-12-mediated antitumor activity Gene Ther 2011 18 5 488 95 21228885 10.1038/gt.2010.170 PMC3125103 Seo SH, Kim KS, Park SH, Suh YS, Kim SJ, Jeun SS, et al. The effects of mesenchymal stem cells injected via different routes on modified IL-12-mediated antitumor activity. Gene Ther. 2011;18(5):488–95. 21228885 10.1038/gt.2010.170 PMC3125103