不同转染方式对腺病毒介导的基因转染效率的影响  

Effect of Different Transfection on Gene Transfer Efficiency Mediated with Adenovirus in Mesenchymal Stem Cells

作　　者：孔霞[1] 郭凌郧[1] 张蕾[1] 郑飞[1] 杨建业[1] 黄永章[1] 唐俊明[1] 王家宁[1,2] 

机构地区：[1]郧阳医学院附属人民医院临床医学研究所,湖北十堰442000 [2]郧阳医学院附属人民医院心内科,湖北十堰442000

出　　处：《郧阳医学院学报》2010年第1期1-4,F0003,共5页Journal of Yunyang Medical College

基　　金：国家自然科学基金(30700306);湖北省卫生厅(JX3B29);湖北省自然基金(2005ABA079);湖北省教育厅(B200624006;200524003)

摘　　要：目的:研究携带绿色荧光蛋白(Green Fluorescent Protein,GFP)基因的重组腺病毒在大鼠骨髓间充质干细胞(MSCs)中的表达。方法:将纯化的GFP重组腺病毒分别用单次转染和多次转染两种不同方法转染MSC,观察其感染效率和GFP表达水平。结果:培养的MSC呈CD90和CD105强阳性,且具有成骨、成脂肪等多向分化能力;携带绿色荧光蛋白基因的重组腺病毒单次转染和多次转染P3代MSC都呈现出浓度依赖性;单次转染在80MOI时转染效率最高,而多次转染在20MOI时转染效率即已达90%。结论:建立稳定、高效表达Ad-GFP的骨髓间充质干细胞实验体系,为MSC的标记及示踪奠定基础。Objective To investigate the expression of recombinant adenovirus encoding green fluorescent protein（GFP） in mesenchymal stem cells （MSCs）. Methods The purified recombinant adenovirus encoding GFP （Ad-GFP） was transfected into MSCs with single or multiple infection, the transfer efficiency of Ad-GFP and the expression level of GFP were measured. Results Cultured MSCs were observed with a multiple differentiation potential into osteoblast ,lipoblast and highly expressed CD105 and CDgO. The Ad-GFP transfeetion of P3 generation MSCs with single or multiple ways were all shown in a concentration-dependent manner. The efficiency of MSCs transfected with Ad-GFP were about 90% in 80 MOI by single infection and in 20 MOI by muhiple infection. Conclusion The system of Ad-GFP expression in MSCs was established successfully and it could provide a basis for further resarch on MSCs labelling and tracing in transplantation experiment.

关 键 词：腺病毒 绿色荧光蛋白 转染 骨髓间充质干细胞 

分 类 号：Q784[生物学—分子生物学]