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

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LIF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。

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

Experiments were performed to determine the factors affecting on culture,clone,isolation and passages of embryonic stem cells(ES) from embryos of Kunmingbai/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)F_1,F_2 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.