Protective Effect of Glutathione Supplementation On Mouse Ovarian Tissue Vitrification and Autologous Transplantation

Ovarian tissue vitrification is a key strategy for fertility preservation, but remains hindered by oxidative damage and compromised tissue viability. This study evaluates the cytoprotective effects of glutathione (GSH) supplementation during the vitrification of mouse ovarian tissue, including autologous transplantation experiments. Mice ovaries were cryopreserved with 0, 2, 4, or 8 mM GSH, followed by comprehensive histological, biochemical, and molecular assessments, as well as autologous transplantation to assess functional recovery. GSH, particularly at 4 mM, significantly preserved follicular architecture, enhanced antioxidant enzyme activity, attenuated mitochondrial dysfunction, DNA damage, and apoptosis, and suppressed pro-inflammatory and fibrotic signaling pathways. GSH also restored angiogenic markers and improved endocrine function in ovarian grafts after autologous transplantation, as evidenced by ameliorative levels of estradiol, progesterone, anti-Müllerian hormone (AMH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH). These findings support GSH as a potent adjuvant for improving ovarian tissue vitrification outcomes and warrant further translational evaluation in fertility preservation protocols.