Efficacy of Human Amniotic Membrane Stem Cell-Conditioned Medium on Improving In Vitro Maturation Efficiency of Mouse Oocytes

Abstract

Highlights What are the main findings? Supplementation of IVM medium with 20% hAMSC-CM significantly increased meiotic maturation rates of mouse oocytes (up to 89.4%) across all PMSG priming regimens (0, 5, 10 IU). 20% hAMSC-CM combined with 5–10 IU PMSG priming markedly enhanced cleavage and blastocyst formation rates, and upregulated IGF1 and BCL2 in resulting blastocysts. What are the implications of the main findings? A dose-optimized, cell-free hAMSC-CM supplement provides a directly transferable, xeno-secretome-based strategy for improving IVM efficiency in assisted reproductive technologies. The mechanistic link between AMH/BMP15/GDF9 cumulus signaling, CASP9 suppression in oocytes, and IGF1/BCL2-mediated blastocyst survival offers new molecular targets to refine IVM media for livestock and clinical fertility preservation. Abstract In vitro maturation (IVM) of oocytes remains less efficient than in vivo maturation, partly because conventional culture media cannot fully reproduce the follicular microenvironment. This study evaluated whether human amniotic membrane stem cell-conditioned medium (hAMSC-CM) improves IVM efficiency of mouse cumulus–oocyte complexes (COCs) recovered from ICR females primed with 0, 5, or 10 IU pregnant mare serum gonadotropin (PMSG). hAMSC-CM was added at 10% or 20% (v/v), and maturation, cleavage, blastocyst formation, and gene expression in MII oocytes, cumulus cells, and blastocysts were assessed. Supplementation with 20% hAMSC-CM significantly increased maturation rates across all PMSG groups, reaching 89.4 ± 1.4% versus 59.5 ± 1.3% in the 5 IU PMSG control (p < 0.05). With PMSG priming, 20% hAMSC-CM also enhanced cleavage and blastocyst formation. Molecular analyses showed suppression of CASPASE9 in MII oocytes, upregulation of AMH, BMP15, and GDF9 with downregulation of AMHR in cumulus cells, and increased IGF1 and BCL2 expression in blastocysts. These findings indicate that 20% hAMSC-CM, together with optimized PMSG priming, improves mouse oocyte IVM efficiency and embryo developmental competence.