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目的:探讨细胞松弛素B在人类体外成熟卵子玻璃化冷冻中的作用。方法:收集常规胞浆内单精子显微注射-胚胎移植(ICSI-ET)周期中未成熟卵母细胞234枚(包括生发泡期即GV期和第1次减数分裂中期即MⅠ期),在体外培养24～48 h,181枚卵母细胞成熟(排出第2极体),随机分成3组并行玻璃化冷冻。A组(43枚)用细胞松弛素B(CB)预处理30 min后行玻璃化冷冻;B组(66枚)用CB预处理20 min后行玻璃化冷冻,C组(72枚)未用CB预处理直接行玻璃化冷冻。D组为对照组30枚体内成熟卵子未用CB处理直接玻璃化冷冻。各组卵子冷冻3周后解冻,复苏的卵子行ICSI辅助授精,观察各组成活率、受精率、卵裂率及囊胚形成率。结果:冻融后的体外成熟卵子,其成活率、受精率、卵裂率及囊胚形成率均显著低于体内成熟卵子(P<0.05)。A组的成活率显著低于B、C两组(P<0.05),A、B、C 3组间的受精率、卵裂率差异无统计学意义(P>0.05),A、B两组未获囊胚,C组仅有1枚囊胚。结论:冻融体外成熟卵子,成活率、受精率、卵裂率均下降,囊胚形成率低,胚胎后期发育潜能显著降低。CB预处理未能提高卵子冷冻的成活率、受精率、卵裂率及胚胎的发育潜能。
Objective: To investigate the role of cytochalasin B in vitrification of human mature oocytes in vitro. Methods: Totally 234 immature oocytes (including GV phase and MⅠ phase of the first meiotic division) were collected during routine intracytoplasmic sperm injection-embryo transfer (ICSI-ET) In vitro culture of 24 ~ 48 h, 181 oocytes matured (excreted second polar body), were randomly divided into three groups parallel vitrification. Group A (43) was pretreated with cytochalasin B (CB) for 30 min before vitrification; Group B (66) pretreated with CB for 20 min followed by vitrification, while group C (72) CB pre-treatment directly to vitrification. Group D, the control group of 30 mature eggs in vivo without CB treatment of direct vitrification. Each group of frozen eggs after 3 weeks of thawing, recovery of eggs ICSI assisted insemination, survival rate, fertilization rate, cleavage rate and blastocyst formation rate were observed. Results: The survival rate, fertilization rate, cleavage rate and blastocyst formation rate of freeze-thawed in vitro mature eggs were significantly lower than those in vivo (P <0.05). The survival rate of group A was significantly lower than that of group B and C (P <0.05), but there was no significant difference in fertilization rate and cleavage rate between A, B and C groups (P> 0.05) No blastocysts, C group only one blastocyst. Conclusion: In vitro maturation of frozen eggs, the survival rate, fertilization rate, cleavage rate are decreased, the blastocyst formation rate is low, embryonic development potential was significantly reduced. CB pretreatment failed to improve the survival rate of eggs frozen, fertilization rate, cleavage rate and embryonic development potential.