影响昆明小鼠和BALB/c小鼠胚胎干细胞分离、克隆、传代的若干因素  

Factors Affecting on Isolation,Clone and Passages of Embryonic Stem Cells of Kunmingbai/str.Mice and BALB/c/str.Mice

作　　者：郭志林[1] 王新庄 王木[3] 张涌[1] 王轶敏 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]河南省肉牛工程技术研究中心,河南郑州450003 [3]河南农业大学牧医工程学院,河南郑州450002

出　　处：《中国兽医学报》2006年第4期448-450,共3页Chinese Journal of Veterinary Science

基　　金：河南省杰出人才创新基金资助项目(0421002100)

摘　　要：对昆明小鼠和BALB/c小鼠2种胚胎的研究表明,2种品系小鼠胚胎均可用作胚胎干细胞(ES)分离建系的材料,两者在ES分离、传代上无显著差异(P>0.05);大鼠心肌细胞条件培养液与添加L IF的ES常规培养液相比,显著提高了小鼠ES细胞F1、F2出现率(P<0.05);采用低浓度消化液使形成ES克隆的比率分别从14%、16%提高到35%、32%,使ES传到第5代的比率分别从1.7%、0提高到5%、7.1%;而采用连续消化法使形成ES克隆的比率从15%、17%提高到40%、50%,使ES传到第5代的比率从0、1%提高到10%、20%;对ES进行伊红染色、核型分析、AKP染色及体外分化能力检测,证实所分离的ES符合小鼠ES的一系列特征。Experiments were performed to determine the factors affecting on culture ,clone ,isolation and passages of embryonic stem cells （ES） from embryos of Kunrningbai/str. mice and BALB/c/str. mice. It was showm that： （1）Both Kunmingbai/ str. mice and BALB/c/str. mice were suitable to get ES cells clones and passages. （2）F1 ,F2 passage rate improved when the medium of rat heart cell-conditioned medium used for ES cells as compared with routine medium added to LIF（P〈0.05）. （3） The low .concentrations of digestion medium used to ES cells improved the rate forming ES clones from 14%, 16% to 35%, 32% and the rate passaged to 5 passages from 1.7% ,0 to 5.0%,7.1%. Continous digestion for ES cells improved the rate forming ES clones from 15%,17% to 40% ,50% and the rate passaged to 5 passages from 0,1% to 10% ,20%. （4）Examined by routine stain, karyotype analysis, AKP strain and the in vitro differentiation capacity, the isolated ES cells bear a series of characters of mice ES cells.

关 键 词：昆明小鼠 BALB/C小鼠 胚胎干细胞 克隆 

分 类 号：S814.9[农业科学—畜牧学]