山羊雄性胎儿生殖细胞建立干细胞系初探

Preliminary studies on establishing stem cell lines from goat male fatal germ cells

体外培养山羊50～68日龄雄性胎儿生殖细胞，并检测它们的碱性磷酸酶（AP）活性和Oct-4蛋白，探讨性别分化后的生殖细胞用于建立干细胞系的可行性及检测指标。当山羊胎儿睾丸细胞体外培养时，生殖细胞及其来源的细胞克隆均呈AP阴性和Oct-4蛋白阴性，其中有部分细胞克隆表现为AP假阳性。山羊胎儿生殖细胞克隆呈隆突状生长，多为圆形，与周围细胞界限分明，但克隆内细胞间界限不清。细胞克隆至少可以培养3代以上。研究结果显示，山羊雄性胎儿生殖细胞可以用于建立生殖系来源的干细胞系；AP和Oct-4蛋白不适宜用来检测体外培养的山羊胎儿生殖细胞及其来源的细胞系。

The goat male fetal germ cells were cultured in vitro and their alkaline phosphatase （AP） activity and Oct-4 protein were detected in order to study the feasibility of establishing stem cell lines from them and the cell line detection methods. When goat male fetal germ cells were cultured on Buffalo rat liver cell feeder layers, the male germ cells and the colonies derived from them all displayed alkaline phosphatase （AP） negative and Oct-4 protein negative, of which some cell colonies displayed AP pseudo-positive. Goat germ cell colonies protruded with growth, most of which were round in shape. They were tightly packed with indistinguishable cell-to-cell boundaries, but had a distinct colony boundary from the surrounding feeder layers. They could be cultured at least up to 3 passages. The results of present study indicate that： （1） goat male fetal germ cells can be employed to establish stem cell lines; （2） establishing stem cell lines from goat male germ cells, AP activity,and Oct-4 protein are unsuitable detection markers.