培养方式对自原始生殖细胞分离克隆小鼠胚胎干细胞的影响

EFFECTS OF CULTURE’S MODE ON ISOLATION AND CLONING OF MOUSE EMBRYONIC STEM-LIKE CELLS

目的 比较与同源胎鼠生殖嵴共培养及其传统饲养层培养的方式 ,从小鼠原始生殖细胞中分离克隆胚胎干细胞的差异。方法 获取 8～ 14d龄的胎鼠 ,制备同源胎鼠成纤维细胞饲养层 ,采用常规饲养层培养的方式和与生殖嵴共同培养的方式 ,分离克隆小鼠胚胎干细胞。结果 在初期分离克隆的过程中 ,采用共同培养的方式优于传统饲养层培养模式。

Objective TO observe the effects of culture’s mode on isolation and cloning of mouse embryonic stem-like cells(ESCs).Methods Primordial germ cells(PGCs) were isolated from 8-14d old mouse fetus.Some of them were cultured on mitotically inactive feeder layer cells,the others and embryonic fibroblast obtained from a conceptus of equivalent gestational age were cocultured.Results During the course of isolating and cloning ES cells,the manner of mouse ES cells and homologous fetus fibroblast coculture was dominant than the one of ES cells cultured depended on mitotically inactive feed layer.The layer cells was provided by mouse embryonic fibroblast or human embryonic fibroblast.Conclusion Isolating and cloning mouse ES cells can btain by co-culture.