牛类胚胎干细胞的分离与克隆  

Isolation and Clone of Bovine Embryonic Stem Like Cells

作　　者：安立龙[1] 杨奇[1] 冯秀亮[1] 杨春荣[1] 雷安民[1] 高志民[1] 窦忠英[1] 邱怀[1] 

机构地区：[1]西北农林科技大学家畜生殖内分泌与胚胎工程农业部重点开放实验室,陕西杨凌712100

出　　处：《中国农业科学》2001年第5期550-555,共6页Scientia Agricultura Sinica

基　　金：国家自然科学基金资助项目 ( 39670 35 9)

摘　　要：选用荷斯坦牛胚胎和小鼠胚胎 ,以 PMOL/ L EF细胞为饲养层 ,以 DMEM+ 15 ml/ 10 0 m l NBS+ 0 .1μm ol/L Na2 Se O3+ 0 .1mm ol/ Lβ-巯基乙醇 + 10 ng/ ml IGF+ 10 0 0 IU/ m l L IF为培养液 ,用消化法 ( ICM用 0 .2 5 g/ 10 0 ml胰蛋白酶 + 0 .0 4 g/ 10 0 ml EDTA) ,ES细胞集落用 ( 0 .12 5 g/ 10 0 ml胰蛋白酶 + 0 .0 2 g/ 10 0 m l EDTA)和机械剥离法离散ICM细胞集落和隆起明显的 ES细胞集落为细胞小块 ,获得了 9个传至 6代的牛类 ES细胞系和 2 2个传至 9代的小鼠类 ES细胞系。从形态特征、组织化学染色、核型分析和 ES细胞分化能力等方面对所分离与克隆的细胞进行鉴定 ,证明其具有The 6 passage bovine ES like cells(12 lines) and 9 passage murine ES like cells(52 lines) were obtained by culturing bovine embryos and murine embryos with tissure medium (DMEM supplemented with 15ml/100ml NBS, 0 1μmol/L Na 2SeO 3, 0 1mmol/L β mercaptoethanol)on the feederlayer of MEF(murine embryonic fibroblast) and disaggregating ICMs and ES cell clones of bovine and murine into smaller cellular fregamentby digestting with 0 125g/100ml trypsin and0 02g/100ml EDTA and scattering with glass needle. The murine ES like cells and bovine ES like cells were identifide in morphological character, histochemistry identification,karyotype analysis and differentiation of ability ES cells. The results showed that they had many characteristics which ES like cells should have.

关 键 词：牛 小鼠 胚胎干细胞 内细胞团 分离 克隆 ES细胞 

分 类 号：S823.36[农业科学—畜牧学]