鼠囊胚在饲养层上的生长行为

Mouse blastocyst growth behavior on feeder layers

目的 比较自然孵化法、去透明带法、免疫外科法去滋养层三种方法对胚胎贴壁率、平均贴壁时间及内细胞团 (ICM)克隆形成率的影响。观察鼠囊胚在小鼠原代胎儿成纤维细胞饲养层上的生长行为。方法 超排卵获得囊胚 ,随机分成A、B、C 3组。A组 :自然孵化法 ;B组 :去透明带法 ;C组 :免疫外科法去滋养层 ,分离获得ICM。观察鼠囊胚及ICM在饲养层上的生长行为。结果 3组胚胎贴壁率、ICM克隆形成率、平均贴壁时间分别为A组 :83%、83%、36～ 4 8h ;B组 :82 %、82 %、2 4～ 36h ;C组 :71 %、71 %、2 4～ 36h。滋养层细胞与ICM同时增殖 ,但滋养层细胞扩展范围小 ,不形成一层半透明膜状结构。ICM在第 4天时能够充分增殖且未分化 ,垂直隆起生长 ,呈典型的卵圆柱状结构。结论 三种方法对胚胎贴壁率、ICM克隆形成率无显著差异 ,去透明带法。

Objective To compare the effect of natural hatching,zona pellucida removal and immunosurgery on mouse blastocyst attachment efficiency,average attachment time and inner cell mass(ICM) cloning efficiency.To observe the mouse blastocyst growth behavior on primary mouse embryonic fibroblast feeder layers (PMEF).Methods Mouse blastocysts were obtained by ovarian hyperstimulation and divided into three groups at random.Group A:natural hatching,group B:zona pellucida removal and group C:trophectoderm cells removal by immunosurgery to take out the ICM.Results Attachment efficiency,ICM cloning efficiency and average attachment time were 83%,83%,36～48h in group A,82%,82%,24～36h in group B and 71%,71%,24～36h in group C respectively.Trophectoderm cells and ICM proliferated together.Trophectoderm cells didn't expand to a wide range and didn't form translucent film like structure on the feeder layers.ICM grew in more rounded clumps before differentiation.Conclusion There is no significant difference between three groups in blastocyst attachment efficiency and ICM cloning efficiency ( P >0.05).The average attachment time is shorter in group B and group C than that in group A.