Tissue engineering and regenerative medicine · 2025
Derivation of Granulosa-Like Cells from Human Endometrial iPSCs for Autologous Estradiol Production.
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Kim Hyun Kyung, Suh Eun Jung, Cho SiHyun, Choi Young Sik, Kim Sinyoung, Park Joo Hyun
Аннотация
BACKGROUND: Granulosa-like cells (GLCs) were differentiated from human endometrium-derived induced pluripotent stem cells (heiPSCs). This study aimed to establish a GLC differentiation protocol as a defined means to produce autologous estradiol (in vitro), highlighting its therapeutic potential as an alternative to conventional menopausal hormone therapy.
METHODS: Endometrial cells from hysterectomy specimens were reprogrammed into iPSCs using episomal vectors encoding SOX2, OCT4, c-MYC, and KLF4. Differentiation was induced using Activin A and CHIR99021 for mesoderm induction, followed by BMP4, Follistatin, and bFGF. Gene expression, flow cytometry, and immunofluorescence were analyzed at each stage. Estradiol production was quantified by ELISA, and its effect on endometrial cell proliferation was evaluated by MTT assay.
RESULTS: Results: The roles of small molecules and growth factors in directing stem cells toward functional chondrocytes are also discussed. Additionally, we briefly examine the emerging integration of artificial intelligence (AI) in cartilage tissue engineering. AI applications such as predicting differentiation outcomes, monitoring chondrogenic progression in real-time, and identifying small-molecule enhancers are poised to accelerate discovery and standardization in the field.
CONCLUSION: GLCs expressing the key markers and capable of E2 production were successfully derived from heiPSCs, which may be developed as a novel source for menopausal hormone therapy.
METHODS: Endometrial cells from hysterectomy specimens were reprogrammed into iPSCs using episomal vectors encoding SOX2, OCT4, c-MYC, and KLF4. Differentiation was induced using Activin A and CHIR99021 for mesoderm induction, followed by BMP4, Follistatin, and bFGF. Gene expression, flow cytometry, and immunofluorescence were analyzed at each stage. Estradiol production was quantified by ELISA, and its effect on endometrial cell proliferation was evaluated by MTT assay.
RESULTS: Results: The roles of small molecules and growth factors in directing stem cells toward functional chondrocytes are also discussed. Additionally, we briefly examine the emerging integration of artificial intelligence (AI) in cartilage tissue engineering. AI applications such as predicting differentiation outcomes, monitoring chondrogenic progression in real-time, and identifying small-molecule enhancers are poised to accelerate discovery and standardization in the field.
CONCLUSION: GLCs expressing the key markers and capable of E2 production were successfully derived from heiPSCs, which may be developed as a novel source for menopausal hormone therapy.