增强型绿色荧光蛋白基因转染小鼠骨髓间充质干细胞的研究
[摘要]目的：以重组腺病毒（Ad）为载体，将增强型绿色荧光蛋白（EGFP）基因转染至小鼠骨髓间充质干细胞（mMSCs）中，为MSCs体内示踪提供实验基础。方法：用全骨髓细胞贴壁法分离纯化小鼠MSCs并体外扩增、鉴定，以重组绿色荧光蛋白基因的腺病毒（Ad-EGFP）转染，并用荧光显微镜和流式细胞仪检测转染率，CCK-8法检测转染细胞的增殖活性。结果：转染后10h MSCs开始表达荧光，6～8天达高峰，转染率可达92.3％，28天仍有表达；转染细胞的增殖活性和未转染细胞无统计学差异。结论：Ad-EGFP能高效、安全的转染mMSCs。
[关键词]骨髓间充质干细胞；增强型绿色荧光蛋白；重组腺病毒；基因转染
[中图分类号]R394.2[文献标识码]A [文章编号]1008-6455（2011）02-0257-04
Study on the transfection of enhanced green fluorescent protein gene into murine marrow mesenchymal stem cells
NING Chang,HU Kai-xun,YU Chang-lin
(Department of Heamatologe,Affiliated Hospital of Academy of Military Medical Science,Beijing 100071,China)
Abstract:ObjectiveTo found the experimental basis of MSCs tracer in vivo,with enhanced green fluorescent protein gene (EGFP)mediated by recombinant adenovirus(Ad) vector transfected murine bone marrow mesenchymal stem cells(mMSCs).MethodsThe mMSCs were separated and eiched by using bone marrow adherent culture and identificated in vitro, to observe the efficiency of transfection of Ad-EGFP by fluorescence microscope and flow cytometry, CCK-8 applied to assay cell proliferation activity.Results10h after transfection MSCs began to express fluorescent, 6 to 8 days reached the peak infection rate of 92.3%, 28 days still expression.Proliferation of transfected cells and untransfected cells no significant difference. Conclusion Ad-EGFP can transfect mMSCs effectively and safly.
Key words:bone marrow mesenchymal stem cells;enhanced green fluorescent protein gene; recombinant adenovirus;gene transfection
骨髓间充质干细胞（bone ma...
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