玻璃化冷冻对人体外成熟卵母细胞微丝分布及发育潜能的影响

Effects of vitrification on microfilaments' distribution and development potentiality of human in vitro matured oocytes

目的通过对比分析人体外成熟卵母细胞玻璃化冷冻前后的微丝分布和发育潜能,探讨冷冻对卵母细胞发育潜能的影响及相关的细胞学机制。方法收集人未成熟卵母细胞,体外培养为成熟卵母细胞。获得的成熟卵母细胞随机分为新鲜组和玻璃化冷冻组,分别行受精和免疫荧光染色分析。受精后观察2PN数、囊胚数及优质囊胚数等指标,染色后用激光共聚焦显微镜观察微丝分布形态。结果玻璃化冷冻复苏后卵母细胞成活率为83.74%。新鲜组2PN数、囊胚数及优质囊胚数均高于玻璃化冷冻组(P<0.05)。新鲜组微丝正常率为71.67%,玻璃化冷冻组为67.50%,两组间差异无统计学意义(P=0.71)。结论适当的玻璃化冷冻复苏方案未改变人体外成熟卵母细胞的微丝分布。但是,冷冻后卵母细胞的发育潜能降低,因此单纯观察微丝分布不能作为冷冻对卵母细胞发育潜能的评价指标。

Objective To find the changes of development potential and microfilament structure which were affected by vitrification for human in vitro matured oocytes.Methods Human in vitro matured oocytes were gathered and random-ly divided into two groups： fresh group and vitrified group.The oocytes were fertilized and immunofluorescencently stained.The numbers of 2PN, blastocyst and high quality blastocyst were calculated and the microfilaments structures were observed by laser confocal microscopy （LSCM）.Results The survival rate of oocytes was 83.74%after vitrifi-cation-warming.The 2PN, blastocyst and high quality blastocyst of the fresh group were significantly higher than those of vitrified group （P＆lt;0.05）.There was no statistical difference in normal microfilaments rate between fresh group and vitrified group （71.67%vs 67.50%, P=0.71）.Conclusion The proper vitrification-warm process doesn’t change the microfilaments structures of human in vitro matured oocytes, but it reduces their development potential.Thus, mor-phology of the microfilaments alone can’ t be taken as the indicator of the developmental potential.