医药前沿
醫藥前沿
의약전연
YIAYAO QIANYAN
2015年
4期
12-13
,共2页
胚胎干细胞%新霉素抗性基因%转基因
胚胎榦細胞%新黴素抗性基因%轉基因
배태간세포%신매소항성기인%전기인
Embryonic stem cells%Neomycin resistance gene%Genetically modified (gm)
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。
目的：建立pOct4-Neo轉基因的小鼠胚胎榦細胞（ESC）株，為進一步研究ESC分化提供有力的保障。方法：設置G418不同濃度進行小鼠ESC培養，確定ESC緻死G418濃度；電穿孔法轉染小鼠ESC，通過G418篩選併挑取暘性剋隆，檢測所得細胞胚胎榦細胞特異性標誌物Oct4及SSEA-1的錶達，通過擬胚體誘導體外分化。結果：ESC的緻死濃度為350μg/mL，在含400μg/mlG418的培養條件下，所得暘性細胞呈剋隆樣鳥巢狀生長，Oct4、SSEA-1錶達暘性，體外可以形成擬胚體和自髮分化。結論：成功對小鼠ESC進行瞭pOct4-Neo的轉基因，為進一步的小鼠ESC體外分化研究奠定瞭基礎。
목적：건립pOct4-Neo전기인적소서배태간세포（ESC）주，위진일보연구ESC분화제공유력적보장。방법：설치G418불동농도진행소서ESC배양，학정ESC치사G418농도；전천공법전염소서ESC，통과G418사선병도취양성극륭，검측소득세포배태간세포특이성표지물Oct4급SSEA-1적표체，통과의배체유도체외분화。결과：ESC적치사농도위350μg/mL，재함400μg/mlG418적배양조건하，소득양성세포정극륭양조소상생장，Oct4、SSEA-1표체양성，체외가이형성의배체화자발분화。결론：성공대소서ESC진행료pOct4-Neo적전기인，위진일보적소서ESC체외분화연구전정료기출。
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.
