【摘要】 目的:评价不同阳离子脂质体介导基因转染血管内皮细胞的转染效率。方法:实验于2006-12/2007-02在中山大学生化实验室及广州市创伤外科研究所完成。采用增强型绿色荧光蛋白基因为报告基因,分别采用Lipofectin、Lipofectamine、Dosper3种不同的阳离子脂质体为载体,转染人脐静脉血管内皮细胞。在24孔板中,每孔加入人脐静脉血管内皮细胞悬液(1×106个细胞),各孔分别加入3种不同阳离子脂质体增强型绿色荧光蛋白质粒复合物,分别于培养24,48h后用荧光显微镜及流式细胞仪测定增强型绿色荧光蛋白在细胞内的表达及转染效率。结果:3种不同阳离子介导的增强型绿色荧光蛋白基因转染的人脐静脉血管内皮细胞内均有绿色荧光蛋白表达,24h后明显,48h后达高峰。Dosper介导组绿色荧光细胞百分比明显高于Lipofectin介导组及Lipofectamine介导组,差异有非常显著意义(P<0.01)。结论:Dosper介导的血管内皮细胞基因转染效率较高,较适合作为血管内皮细胞的基因转染载体。更多还原

【Abstract】 AIM:To evaluate the efficiency of different types of cationic liposome-mediated gene transfection to culture human umbilical vein endothelial cells(HUVEC).METHODS:The experiment was performed at the Laboratory of Biochemistry of Sun Yat-sen University and Guangzhou Institute of Traumatic Surgery from December 2006 to February 2007.Reporter gene expressing enhanced green fluorescent protein(EGFP)was used in order to evaluate transfection efficiency.HUVEC were transfected with Lipofectin,Lipofectamine and Dosper in 24-well plate.HUVEC suspension(1×106 cells)was added in each well,and Lipofectin,Lipofectamine and Dosper combined with EGFP compound were added.At hours 24 and 48,EGFP expression and transfection efficiency were determined by fluorescent microscope and flow cytometer.RESULTS:EGFP in HUVEC by different types of cationic liposome-mediated gene transfection was found,and their expression increased 24 hours after transfection with a summit at hour 48.The efficiency of Dosper mediated gene transfection group was higher than that in the Lipofectin mediated gene transfection group and Lipofectamine mediated gene transfection group(P < 0.01).CONCLUSION:The transfection efficiency of Dosper mediated gene transfection to HUVEC is higher and suitable for gene transfection to HUVEC as a gene transfection vehicle.更多还原