the beneficial effects of antifreeze proteins in the vitrification of immature mouse oocytes的有利影响抗冻蛋白在未成熟小鼠卵母细胞玻璃化.pdf

The Beneficial Effects of Antifreeze Proteins in the Vitrification of Immature Mouse Oocytes Jun Woo Jo1,2,4., Byung Chul Jee1,2., Chang Suk Suh1,2*, Seok Hyun Kim2,3 1 Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea, 2 Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea, 3 Department of Obstetrics and Gynecology, Seoul National University Hospital, Seoul, Korea, 4 Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University Seoul, Korea Abstract Antifreeze proteins (AFPs) are a class of polypeptides that permit organismal survival in sub-freezing environments. The purpose of this study was to investigate the effect of AFP supplementation on immature mouse oocyte vitrification. Germinal vesicle-stage oocytes were vitrified using a two-step exposure to equilibrium and vitrification solution in the presence or absence of 500 ng/mL of AFP III. After warming, oocyte survival, in vitro maturation, fertilization, and embryonic development up to the blastocyst stage were assessed. Spindle and chromosome morphology, membrane integrity, and the expression levels of several genes were assessed in in vitro matured oocytes. The rate of blastocyst formation was significantly higher and the number of caspase-positive blastomeres was significantly lower in the AFP-treated group compared with the untreated group. The proportion of oocytes with intact spindles/chromosomes and stable membranes was also significantly higher in the AFP group. The AFP group showed increased Mad2, Hook-1, Zar1, Zp1, and Bcl2 expression and lower Eg5, Zp2, Caspase6, and Rbm3 expression compared with the untreated group. Supplementation of the vitrification medium with AFP has a protective effect on