两种来源小鼠胚胎干细胞的分离与培养

Isolation and cultivation of mouse embryonic stem cells from two sources

目的 :从小鼠早期胚胎和原始生殖细胞中分离和培养胚胎干细胞 (ES细胞 )。方法 :取小鼠 3 .5天胚龄的囊胚 ,培养在小鼠胚胎成纤维细胞饲养层上 ,3～ 4天后分离出内细胞团再培养 ;取 1 3 .5天胚龄胚胎的生殖嵴组织 ,分离出原始生殖细胞 ,在小鼠胚胎成纤维细胞饲养层培养 ;观察两种来源的细胞集落生长行为 ,并用AKP染色鉴定细胞集落。结果 :两种来源的细胞集落均具有ES细胞鸟巢状典型特征 ,AKP染色阳性。结论 :两种来源的小鼠胚胎干细胞均可分离和培养。

Objective:To establish a method to culture mouse embryonic stem cells from embryos and primordial germ cells(PGCs).Methods:The 3.5 days old embryos were taken out and cultured on the mouse embryonic fibroblast feeder cells for 3 to 4 days,and the cells from inner cell mass(ICM) were isolated and further cultured.The genital cells were isolated from the genital crest of 13.5 days old embryos and cultured on the embryonic fibroblast feeder cells.The stem cells from the two resources and their colony were identified by characterized morphology and AKP staining.Results:The stem cells from both resources were grown and possess characterized as nest like in morphology and AKP staining as positive.Conclusions:The stem cells from both resources were successfully isolated and cultured in this experiment.