昆明小鼠ES细胞的分离培养及鉴定  

Isolation Culture and Identification of ES Cells in Kunming Mouse

作　　者：张廷龙[1] 董雅娟[1] 柏学进[1] 赵晶[1] 岳福杰 史文升[1] 龚宜超[1] 

机构地区：[1]青岛农业大学动物胚胎工程中心,山东青岛266109 [2]荣成市出入境检验检疫局,山东荣成265300

出　　处：《安徽农业科学》2008年第13期5380-5383,5401,共5页Journal of Anhui Agricultural Sciences

基　　金：青岛市科技将才项目(2005620313)

摘　　要：[目的]摸索一种适合昆明小鼠ES细胞分离培养的方法,以提高昆明小鼠ES细胞的建系率。[方法]分别采用5种方法分离ICM和ES细胞集落,摸索了以BMSCs作为饲养层时,丝裂酶素处理的合适时间。[结果]连续消化法要明显好于其他4种方法。MMC处理BMSCs1、1.5、2h时效果较好,3个时间点无显著差异（P〉0.05）。mMEF作为饲养层与BMSCs作为饲养层时获得5代ES细胞相比差异不显著（P〉0.05）。[结论]连续消化法分离ES细胞效率较高,饲养层采用MMC处理1～2h的BMSCs饲养层或常规mMEF饲养层对昆白小鼠ES细胞无明显影响。[Objective] The purpose was to grope a method suitable for the isolation culture of ES cells in Kunming mouse so as to enhance their line-building rate.[Method] ICM and ES cell colonies were isolated by 5 methods separately and the suitable treating time of mitomycin C（MMC） with BMSCs as feeder layer was found out.[Result] The continuous digestion was better than the other 4 methods significantly.The effects of treating BMSCs by MMC for 1,1.5 and 2.0 h were better and there was no significant difference among the three time points（P〉0.05）.With mMEF and BMSCs as feeder layers,among the obtained 5th generation ES cells,there was no significant difference（P〉0.05）.[Conclusion] The continuous digestion was higher efficient in isolating ES cells.Whether the feeder layer was BMSCs treated by MMC for 1～2 h or was the conventional mMEF,it had no significant influence on ES cells in Kunming mouse.

关 键 词：昆明小鼠 KS细胞 BMSCS ICM 

分 类 号：S865.13[农业科学—野生动物驯养]