The Secretome of menstrual blood-derived endometrial stem cells ameliorates chemotherapy-induced ovarian injury by inhibiting necroptosis via the HSPA8-ATRIP axis.

Chemotherapy-induced ovarian injury poses a significant reproductive health challenge for young female cancer survivors, frequently resulting in premature ovarian insufficiency (POI) and infertility. This study demonstrates that menstrual blood-derived endometrial stem cells (MenSCs) and their conditioned secretome (MenSC-S) effectively ameliorate cisplatin-induced ovarian atrophy and functional decline, restoring primordial and growing follicle reserves and rescuing the expression of key functional biomarkers, including anti-Müllerian hormone (AMH), follicle-stimulating hormone receptor (FSHR), estrogen receptor alpha (ERα), and aromatase (Cyp19a1). Mechanistically, we show that chemotherapy induces a pathological shift toward necroptosis in ovarian granulosa cells (GCs) through activation of the RIPK1/RIPK3/p-MLKL signaling cascade, concomitant with suppression of the endogenous necroptosis inhibitor caspase-8. We identify heat shock protein family A member 8 (HSPA8) as an indispensable paracrine mediator within MenSC-S that directly interacts with ATR-interacting protein (ATRIP) to establish a previously unrecognized regulatory axis. This physical HSPA8-ATRIP interaction is associated with reduced intracellular reactive oxygen species (ROS) accumulation and restoration of caspase8-mediated inhibition of necroptosis, thereby attenuating the necroptotic signaling cascade. Collectively, our findings support a cell-free therapeutic strategy that achieves efficacy comparable to MenSCs transplantation while offering enhanced biosafety and manufacturing standardization, thereby providing a robust theoretical and experimental foundation for the clinical translation of MenSC-based interventions in oncofertility.