不同转染方法及GFP表达载体对小鼠胚胎干细胞转染效率的比较

Effect of Different Transfection Methods and Green Fluorescent Protein Expression Vectors on Transfection Efficiency in Mouse Embryonic Stem Cell

目的对比不同的转染方法及绿色荧光蛋白(GFP)表达载体对小鼠胚胎干细胞(ESC)的转染效率,为进一步研究ESC及衍生细胞移植示踪提供工具。方法用阳离子脂质体转染小鼠胚胎干细胞系ES-D3,对比贴壁细胞转染法、悬浮细胞转染法及不同载体pCX-EGFP、p-EGFP-C1和pIRES-hrGFP的转染效率。结果悬浮法和贴壁法转染效率分别为(90.0±4.1)%,(70.0±2.3)%,两组比较差异有显著性意义(P<0.05)。3种载体转染效率分别为pCX-EG-FP(90.0±4.1)%,pIRES-hrGFP(32.0±6.0)%,p-EGFP-C1(4.0±0.4)%。3组间两两比较差异均有显著性意义(P<0.05)。结论悬浮转染法转染pCX-EGFP可获得较高的转染效率。

Objective To compare the effects of different transfection methods and green fluorescent protein （GFP） expression vectors on transfeetion efficiency in mouse embryonic stem cells and offer a tool /or transplanted cell tracking in vivo. Methods ES-D3 was transfeeted with lipofectamine 2000 and transfection efficiency was compared between suspension and ad herent transfection and different transfection vectors such as pCX-EGFP, p-EGFP-C1 , pIRES-hrGFP protein in mouse embryonic stem cells. Results Transfection efficiency by suspension method and adherent method was （90.0±4.1） % and （70.0± 2.3） % separately with the difference being statistically significant （P〈0.05）. Transfection efficiency of pCX-EGFP, pIRES- hrGFP and p-EGFP-C1 was （90.0±4.1） %, （32.0±6.0） % and （4.0±0.4） % separately with the difference being significant among them （P〈0.05）. Conclusion Transfection of mouse embryonic stem cells with suspension method and pCX-EGFP could obtain higher transfection efficiency of the GFP in mouse embryonic stem cells.