利用细胞饲养层分离培养大鼠胚胎干细胞

Isolation and Cultivation of Mouse Embryonic Stem Cell

目的 从大鼠的早期胚胎分离和培养胚胎干细胞.方法收集大鼠的囊胚,将其培养在小鼠胚胎成纤维细胞饲养层上,5～6d后取隆起生长的内细胞团块分离后再培养,观察集落的生长情况,并在光镜下观察细胞形态、分化过程及细胞核型的检测.结果细胞集落性生长,符合胚胎干细胞的特征.结论大鼠囊胚在胚胎成纤维细胞饲养层上可发育成胚胎干细胞,并能进行传代.

Objective To isolate and culture mouse embryonic stem cell from embryos of mouse. Methods The blastocytes of 5d from mouse, culture on the mouse fibroblaste ceil feeder layers for 5 - 6 days, and the ceils from inner cell mass （ICM） were isolated and subsequently cultured in vitro. The stem ceil were identified by characterized morphology, alkaline phosphatase activity assay, and ceil karyotypoes examination. Results The stem ceils were grown and possess characterized morphology, and the stem ceil were sat- isfied all traits of mouse ES cell line. Conclusions The stem ceil from mouse could be culture and isolated in the mouse fibroblaste cell feeder layers.