Comparison of oxidative status of mouse pre-antral follicles derived from vitrified whole ovarian tissue and vitrified pre-antral follicles in the presence of alpha lipoic acid

摘要

Aim The main goal of this study was to compare developmental competence and oxidative status of vitrified-warmed pre-antral follicles (VPF) with pre-antral follicles derived from vitrified-warmed ovarian tissue (VOF) in the presence of alpha lipoic acid (ALA). Materials and Methods Ovarian tissue and isolated pre-antral follicles were exposed to equilibration solution and then vitrification solution. After thawing of LN2 snap-frozen samples, pre-antral follicles were cultured with or without ALA for 12 days that followed by hCG-induced ovulation. MII oocytes were in vitro fertilized and embryo cleavage assessed. Reactive oxygen species (ROS) and total antioxidant capacity (TAC) levels of cultured pre-antral follicles were measured. Results The rates of survival, antral-like cavity formation, MII oocytes, fertilization, 2-cell embryo and blastocyst development were higher in VPF compared to VOF. These rates were higher in ALA-supplemented groups in comparison to their respective groups. An increase and a decrease in ROS production and TAC levels were observed up to the 96h during cultivation period, respectively. ROS level was lower in cultured VPF compared to VOF. In ALA-treated groups, ROS level decreased to reach comparable values of starting point and TAC levels increased after 24h of culture and then remained constant. Conclusion Developmental outcomes showed vitrification of pre-antral follicles is more appropriate method than that of whole ovarian tissue. Moreover, it seems that inclusion of ALA improved in vitro development of pre-antral follicles in both vitrified and non-vitrified samples.