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

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.