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病毒及非病毒载体对U937单核细胞的转染效果 被引量：3

Transfection efficiency of virus and non-viral vectors against U937 monocytes
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摘要 目的:比较研究腺病毒、慢病毒和Lipofectamine 2000 3种方法转染U937细胞的效果差异,以寻找一种简便、高效转染U937细胞的方法。方法:分别采用Lipofectamine 2000介导质粒载体(pGPHI/GFP/Neo),腺病毒(Adv-GFP-NC)和慢病毒(LV-GFP-NC)3种方法转染U937细胞,于脂质体转染48 h后、腺病毒和慢病毒转染96 h后在荧光显微镜下观察绿色荧光蛋白(green fluorescence protein,GFP)表达情况,并收集细胞以流式细胞仪检测GFP阳性细胞的比例。台盼蓝染色法计数不同转染方法对细胞活力的影响。结果:Lipofectamine 2000介导质粒载体转染组、腺病毒转染组GFP阳性细胞极少,流式细胞仪检测GFP阳性细胞均<4%;慢病毒转染组GFP阳性细胞满视野,感染复数=100时,GFP阳性细胞约占90%。台盼蓝染色法显示腺病毒、慢病毒组活细胞率明显高于脂质体组,差异具有统计学意义(P<0.01)。结论:慢病毒不仅使Lipofectamine 2000和腺病毒能有效地将外源基因导入U937细胞内表达,也是转染U937细胞的理想载体。 Objective：To compare the transfection efficiencies among adenovirus,lentivirus and Lipofectamine 2000 and to explore an efficient way for U937 cells transfection. Methods： U937 cells were transfected with Lipofectamine 2000-plasmid（pGPHI/GFP/Neo）, adenovirus vector（Adv-GFP-NC） and lentivirus vector（LV-GFP-NC）, respectively. Expressions of green fluorescence protein（GFP） were detected under fluorescence microscopy respectively at 48 h after Lipofectamine 2000 transfection, at 96 h after adenovirus vector （Adv-GFP-NC） transfection and at 96 h after lentivirus vector transfection. U937 cells were collected for GFP-positive cells count with flow cytometry. Meanwhile,Trypan blue staining was employed to determine the cell viability. Results：GFP-positive cells were rare in Lipofectamine 2000 group and adenovirus vector group, with percentages of GFP-positive cells less than 4%. But in lentivirus group, the percentage of GFP-positive cells was about 90%（multiplicity of infection= 100）. Trypan blue staining showed that the ratios of living cells were significantly higher in adenovirus group and lentivirus group than in Lipofectamine 2000 group（P〈0.O1 ）. Conclu sions：Lentivirus, other than Lipofectamine 2000 or adenovirus, can efficiently transfect exogenous genes into U937 cells for gene exnression and it is an ideal vehicle for U937 cell trandfection
作者 张艳君 贾秀红 李建厂 徐酉华
机构地区 滨州医学院附属医院儿科.滨州 重庆医科大学附属儿童医院肿瘤实验室.重庆
出处 《重庆医科大学学报》 CAS CSCD 北大核心 2013年第2期161-164,共4页 Journal of Chongqing Medical University
基金 山东省科学技术发展计划资助项目(编号:2010GSF10264)
关键词 U937 腺病毒 慢病毒 质粒 转染效率 U937 adenovirus lentivirus plasmid transfeetion efficiency
分类号 R725.5 [医药卫生—儿科]
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参考文献16

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共引文献11

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重庆医科大学学报
2013年 第2期

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