病毒及非病毒载体对U937单核细胞的转染效果

Transfection efficiency of virus and non-viral vectors against U937 monocytes

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。

Abstract:

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<0.01）. Conclu－sions：Lentivirus，other than Lipofectamine 2000 or adenovirus，can efficiently transfect exogenous genes into U937 cells for gene ex－pression and it is an ideal vehicle for U937 cell transfection.