重组腺病毒介导的绿色荧光蛋白基因转染大鼠间充质干细胞的实验研究

目的 体外研究重组腺病毒载体介导绿色荧光蛋白基因转染大鼠间充质于细胞(MSCs)的方法.方法 用密度梯度离心及贴壁法相结合的方法分离纯化大鼠MSCs并进行鉴定,重组绿色荧光蛋白的腺病毒载体Ad-GFP转染,荧光显微镜检测其转染效率,CCK-8法检测转染细胞的增殖能力.结果 Ad-GFP对大鼠MSCs的感染率高达90.0%,感染1月后仍有稳定表达;转染细胞的增殖能力与未转染细胞相比,差异无统计学意义(P＞0.05).结论 重组腺病毒载体Ad-GFP可高效感染MSCs,基因转染后细胞的增殖能力不受影响,可作为一种高效的大鼠MSCs的标记方法.

Green fluorescent protein gene transfection mediated by recombinant adenovirus vector to rat bone marrow mesenchymal stem cells

Objective To investigate the efficient transfection of green fluorescent protein gene(GFP)mediated by recombinant adenovirus vector(Ad-GFP)to rat bone marrow mesenchymal stem cells(MSCs)in vitro.Methods Wistar rat bone marrow-derived MSCs were separated and purified in vitro by Percoll density gradient centrifugation combined with adherent cell culture followed by identification with flow cytometry.MSCs infected by Ad-GFP were observed and the transfection efficiency was assessed by fluorescence microscope.The proliferative ability of these cells was tested by CCK-8.Results The transfection efficiency was as high as 90.0%.Expression of GFP gene of infected MSCs was stable for 1 month after infection.There was no statistically difference in proliferative ability between the infected MSCs and non-infected ones(P>0.05).Conclusions The infected MSCs with Ad-GFP expressed GFP with high efficiency and retain the ability of proliferation as non-infected MSCs.Transgection with Ad-GFP is a highly effective method for labeling MSCs.