小鼠胚胎干细胞转OCT4启动的新霉素抗性基因的研究  

Embryonic stem cells in mice OCT4 start neomycin resistance gene research

作　　者：张爱霞[1,2] 姚琼凤[1] 赵书策[1] 

机构地区：[1]嘉应学院生命科学学院,广东梅州514015 [2]中山大学中山医学院干细胞与组织工程中心,广东广州510080

出　　处：《医药前沿》2015年第4期12-13,共2页Journal of Frontiers of Medicine

摘　　要：目的：建立pOct4-Neo转基因的小鼠胚胎干细胞（ESC）株，为进一步研究ESC分化提供有力的保障。方法：设置G418不同浓度进行小鼠ESC培养，确定ESC致死G418浓度；电穿孔法转染小鼠ESC，通过G418筛选并挑取阳性克隆，检测所得细胞胚胎干细胞特异性标志物Oct4及SSEA-1的表达，通过拟胚体诱导体外分化。结果：ESC的致死浓度为350μg/mL，在含400μg/mlG418的培养条件下，所得阳性细胞呈克隆样鸟巢状生长，Oct4、SSEA-1表达阳性，体外可以形成拟胚体和自发分化。结论：成功对小鼠ESC进行了pOct4-Neo的转基因，为进一步的小鼠ESC体外分化研究奠定了基础。ObjectiveEstablish pOct4 Neo-genetically modified mice embryonic stem cells （ESC） strains, ESC differentiation provide strong guarantee for further study. Methods Mice ESC training set different concentration of G418, determine the ESC lethal concentration of G418; Transfection mice ESC electroporation method, by G418 screening and take positive clones, cells, embryonic stem cells measured Oct4 tumor-specific markers and the expression of SSEA 1, through the embryoid bodies inducing differentiation in vitro.ResultsESC lethal concentration is 350 mu g/mL, in the land of 400 mu g/mL G418 cultivation condition, the positive cells were cloned sample nest, Oct4, SSEA 1 express positive, can form embryoid bodies and the spontaneous differentiation in vitro. ConclusionsSuccess on the mice ESC pOct4 Neo-genetically modified （gm）, in order to further the mice ESC differentiation in vitro research laid the foundation.

关 键 词：胚胎干细胞 新霉素抗性基因 转基因 

分 类 号：R392[医药卫生—免疫学]