小鼠胚胎生殖细胞体外培养

THE CULTURE OF STEM CELLS FROM MURINE EMBRYO IN VITRO

目的:研究胚胎生殖细胞的体外生长条件和分化潜能,探索从胚胎生殖细胞获得皮肤细胞的可能性。方法:在解剖显微镜下分离取出10天龄的胎鼠生殖嵴,消化法得到原生殖细胞,接种在胎鼠的成纤维细胞饲养层上,培养基使用α-MEM添加药用级的bFGF,或者添加成纤维细胞的上清液,配制成条件培养基。培养的细胞用形态学观察法,碱性磷酸酶染色法和诱导分化的方法来鉴定。结果:初步摸索出胚胎生殖细胞体外培养的简单方法,验证了它的分化潜力,观察到了神经细胞、心肌细胞、和表皮样细胞。结论:本试验为简易条件下的胚胎生殖细胞的研究做出有益的探索,为大规模获得表皮细胞提供了一个新的思路。

Objective:In order to investigate the required culture condition and the ability to differentiation of the embryonic stem cells,and to try the possibility that obtaining the skin cells from the embryoni c stem cells.Methods:Under the anatomy microscope,we obtained genital ridge from the 10th embryonic murine,and got primordia l germ cells with trypsin,then feed t hem on the feeder cells made of murine embryonic fibroblast.The culture medium is composed of normalα-MEM and bFGF of drug class,or the con ditioned medium with the supernatant from emb ryonic murine fibroblast.The cultu red cells from embryo was watched thr ough morphology.To i-dentified the cells,we used alkali phosphatase to stain,and induced them to differentiate to all three layer cells of embryonic.Results:A simple method is approximately obt ained to culture the stem cells in vit ro,to test their differentiation ab ility,and to observed neuro-cyte,cardiac muscle cell,and epider -mic cell.Conclusion:A favorite investigation has been done by our experiment for studying the embryonic stem cells in a simple way,and it supplys a new method to get epid ermic cells at a large scale.[