摘要
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.
出处
《华中科技大学学报(医学版)》
CAS
CSCD
北大核心
2006年第4期536-537,541,共3页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30370505)
关键词
绿色荧光蛋白
胚胎干细胞
转染
green fluorescent protein
embryonic stem cell
transfection