【摘要】 目的:比较3种玻璃化冷冻对小鼠生发泡期卵母细胞的存活及胚胎发育潜能的影响。方法:将小鼠生发泡期卵丘复合物(n=175、177、190)和裸卵(n=150、149、150)分别随机分为3组:采用一步乙二醇法、乙二醇加二甲基亚砜法、三步乙二醇法进行冷冻保存。解冻存活体外培养成熟后行卵胞浆内单精子显微注射受精,进一步评价胚胎发育潜能。结果:冻融小鼠生发泡期卵丘复合物,三步乙二醇法与一步乙二醇法的存活率、成熟率、受精率均高于乙二醇加二甲基亚砜组(P均<0.05);三步乙二醇法的优质胚胎和囊胚率均高于一步乙二醇法和乙二醇加二甲基亚砜法(P均<0.05)。冻融小鼠生发泡期裸卵,三步乙二醇法的存活率及三步乙二醇法、一步乙二醇法的受精率及卵裂率均高于乙二醇加二甲基亚砜法(P均<0.05)。三步乙二醇法中,卵丘复合物组的存活率、成熟率、受精率、卵裂率及优质胚胎率均高于裸卵组(P均<0.05)。结论:3种玻璃化冷冻中,三步乙二醇法可以获得高质量的冻融小鼠生发泡期卵母细胞,是一种理想的冷冻方法;带完整卵丘颗粒细胞的生发泡期卵母细胞的冻融效果更佳。更多还原

【Abstract】 Aim:To explore the effects of three vitrification methods on survival and development competence of the germinal vesicle(GV) stage mouses oocytes.Methods:The GV stage mouses COCs(n=175,177,and 190) and DO(n=150,149,and 150) were cryopreserved by three vitrification methods:one-step ethylene glycol,EG plus dimethyl sulfoxide,and three-step EG.After being thawed,the survival oocytes extrusing the polar body I in vitro maturation for 36 h were fertilizated by ICSI respectively,and embryo developmental competence was assessed.Results: The survival rate,matured rate,and cleavage rate of three-step EG and one-step EG increased significantly as compared with EG plus DMSO method(P<0.05).Good quality rate and blastocyst rate of three-step EG increased significantly as compared with one-step EG and EG plus DMSO methods(P<0.05).The survival rate of three-step EG,the fertilization rate of three-step EG and one-step EG increased significantly as compared with EG plus DMSO method(P<0.05).Compared with DO group,the survival rate,maturation rate and fertilization rate,cleavage rate and good quality rate of COCs group increased significantly(P<0.05).Conclusion: Three-step EG method is a better freezing method,which is an ideal method for cryopreserving the GV mouse oocytes.And cryopreserving COCs has a better result.更多还原