原始生殖细胞是用作分离和克隆胚胎干细胞的一种新的细胞资源。将30~45日龄的蒙古绵羊胚胎生殖嵴及其临近组织用机械剪碎和胰蛋白酶+EDTA消化处理,添加DMEM(低糖)+10%FBS(犊牛血清)、38.0℃、5%CO2和饱和湿度条件下进行培养。其结果:未加任何细胞生长因子的情况下与其胎儿成纤维细胞共培养的方式也能分离得到类胚胎干细胞集落。这些集落细胞经多次克隆传代具有胚胎干细胞的诸多特征,如:具有连续传代的能力,细胞集落有典型鸟巢状结构,AKP染色呈阳性,核型分析结果染色体正常等。这些表明该细胞具有多能性,是绵羊类ES细胞。
The primitive germ cell is a kind of new cell resources used as isolation and coloning embryonic stem (ES)cell . Genital ridge and the surrouding tissue from 30 ~ 45 day old fetuses of Mongolia sheep were minced with the machine and disaggregated by trypsin/EDTA solution, and then , PGCS and it's fetal fibroblast were co-cuhured at 38℃ ,5% CO2 concentration and saturated humidity in a CO2 incubator. Dublecco' s Modified Eagle's Medium (DMEM)with 10% fetal bovine serm (FBS) was used as culture medium without any cell growth factors.