| 慢病毒载体介导绿色荧光蛋白基因转染血管内皮祖细胞的实验研究 |
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| 引用本文： | 宋淑敏,刘铭,彭强,植勇,杨春. 慢病毒载体介导绿色荧光蛋白基因转染血管内皮祖细胞的实验研究[J]. 山东医药, 2011, 51(4): 27-29 |
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| 作者姓名： | 宋淑敏 刘铭 彭强 植勇 杨春 |
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| 作者单位： | 泸州医学院附属医院,四川泸州,646000 |
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| 基金项目： | 国家自然科学基金项目资助项目(30700876); 四川省教育厅项目(2006B026) |
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| 摘 要： | 目的探讨HIV-1来源的慢病毒载体介导绿色荧光蛋白（GFP）基因转染血管内皮祖细胞（EPCs）的可行性和方法。方法用梯度密度离心法分离人脐带血内皮祖细胞,在EGM-2培养基中培养。用细胞免疫荧光染色和流式细胞仪检测其表达情况。以HIV-1来源的慢病毒为载体、以GFP基因为目的基因转染EPCs,MTT法检测不同病毒滴度（MOI）时细胞增殖情况并观察转染率。结果单个核细胞经EGM-2培养基培养1周后即分化成EPCs。GFP转染后48 h细胞即发出绿色荧光。MOI 1∶10转染组细胞转染率低于MOI 1∶50组（P〈0.05）。MOI 1∶50转染后的细胞与未转染GFP组比较,生长曲线无明显差异（P〉0.05）。MOI 1∶100组转染后细胞的增殖处于停滞状态。结论采用HIV-1来源的慢病毒载体介导GFP基因转染标记EPCs是可行的。以MOI 1∶50进行转染对细胞生长影响小,转染效率高。
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| 关 键 词： | 血管内皮祖细胞 转染 慢病毒 绿色荧光蛋白质类 |
Study of green fluorescent protein gene transfection in endothelial progenitor cells mediated by lentivirus vector |
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| SONG Shu-min,LIU Ming,PENG Qiang,ZHI Yong,YANG Chun. Study of green fluorescent protein gene transfection in endothelial progenitor cells mediated by lentivirus vector[J]. Shandong Medical Journal, 2011, 51(4): 27-29 |
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| Authors: | SONG Shu-min LIU Ming PENG Qiang ZHI Yong YANG Chun |
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| Affiliation: | SONG Shu-min,LIU Ming,PENG Qiang,ZHI Yong,YANG Chun(The Affiliated Hospital of Luzhou Medical College,Luzhou 646000 P.R.China) |
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| Abstract: | Objective To understand the feasibility and methods of green fluorescent protein(GFP) transfection in endothelial progenitor cells(EPCs) mediated by lentivirus vector HIV-1.Methods EPCs harvested from umbilical cord blood were isolated by gradient density centrifugation and cultured in EGM-2 medium.The cells were identified by immunofluorescence staining and flow cytometry.Cell proliferation and transfection efficiency were assayed by MTT method.Results The mononuclear cells cultured in EGM-2 medium were co... |
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| Keywords: | endothelial progenitor cells transfection lentivirus green fluorescent proteins |
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