山羊PGCs用于分离与克隆类ES细胞  

Isolation and Cloning of Goat ES-like Cells from PGCs

作　　者：韩建永[1] 桑润滋[1] 孙国杰[1] 王志刚[1] 李俊杰[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001

出　　处：《中国兽医学报》2002年第4期344-347,共4页Chinese Journal of Veterinary Science

基　　金：河北省自然科学基金资助项目 (3 0 2 2 89) ;河北省"十五"重大科技攻关资助项目 (0 12 2 0 40 7D)

摘　　要：选择健康成年本地白山羊 ,自然发情、配种后 4 4 d取胎儿 ,以传统的原始生殖细胞 (PGCs)分离与克隆的方法和PGCs与其胎儿生殖嵴周围组织细胞共同培养的方法获得类胚胎干细胞 (类 ES细胞 ) ,并对山羊类 ES细胞在不同饲养层上进行培养。结果表明 ,采用传统方法和共培养的方法并添加细胞因子均能分离获得类 ES细胞。分离获得的类ES细胞在同源 (山羊 )胎儿细胞饲养层上生长效果较好 ,可传 4代或 5代 ,而在小鼠原代成纤维细胞饲养层上类 ES细胞仅传 3代。另外 ,共培养不添加细胞因子组仅获 1个类 ES细胞集落 。ES like cells of goat were isolated by the traditional method of isolation and cloning of primordial germ cells (PGCs) and the method of co culture of PGCs and the cells surrounding the gonadal anlage, from day 44 fetuses of healthy local white goat after natural estrus and mating. These ES like cells were cultured on different feeders. The results indicated that ES like cells were both derived from the traditional method and co culture in medium with cell growth factors. The ES like cells on primary homologous(goat) embryonic fibroblasts(PGEF) feeder grew better than those on primary mouse embryonic fibroblast(PMEF), which were observed for 4 5 passages and 3 passages respectively. However, only one ES like cell colony was derived by co culture in medium without cell growth factor and was lost after passage.

关 键 词：分离 克隆 山羊 胚胎干细胞 原始生殖细胞 类ES细胞 

分 类 号：S827.3[农业科学—畜牧学]