外源基因转染血管内皮细胞条件优化  

Optimization of exogenous gene transfection in human umbilical vein endothelial cells

作　　者：章倩倩[1] 丁一[1] 刘红英[1] 刘翼龙[1] 陈胜霞[1] 胡曦文[1] 王丽京[1] 

机构地区：[1]广东药学院血管生物学研究所,广东广州510006

出　　处：《广东药学院学报》2013年第3期310-313,共4页Academic Journal of Guangdong College of Pharmacy

基　　金：国家自然科学基金(31200896);广东省医学科研基金(B2012181);广东省自然科学基金(S2012040007658)

摘　　要：目的探讨阳离子脂质体及电穿孔法介导外源基因转染血管内皮细胞的转染效率并进行条件优化。方法采用绿色荧光蛋白基因为报告基因,分别采用Lipofectamine^(TM)2000阳离子脂质体为载体以及电穿孔方法,转染人脐静脉血管内皮细胞(HUVEC)。培养48 h后,用荧光显微镜观察绿色荧光蛋白在HUVEC内的表达及转染效率。结果 Lipofectamine^(TM)2000阳离子脂质体介导组有少量细胞可见弱荧光信号,而电穿孔法转染组可见较强荧光信号,其中以电击条件为电场强度1100 V、脉冲时间20 ms、电击2次的转染效率最好。结论电穿孔法介导的血管内皮细胞基因转染效率较高,可作为血管内皮细胞的外源基因转染的方法。Abstract： Objective To compare the efficiency of cationic liposome- and eleetroporation-mediated exogenous gene transfeetion in human umbilical vein endothelial cells （HUVEC）, and optimize the transfection condition. Methods HUVEC cells were transfeeted with plasmid DNA, carried green fluorescent protein （GFP） gene, by LipofectaminTM 2000 or electroporation. The GFP expression and transfection efficiency were examined by fluorescent microscope at 48 h after transfection. Results The hypofluorescence signal was observed in the LipofectamineTM 2000-treated group, however the hyperfluorescenee signal was detected in the electroporation group. The optimal condition in the electroporation group included the electric field strength I 100 V,pulse time 20 ms and electric shock 2 times. Conclusion The efficiency of exogenous gene transfeetion with electroporation is higher than cationic liposome,which is suitable for exogenous gene transfection in HUVEC cells.

关 键 词：人脐静脉血管内皮细胞 基因转染 阳离子脂质体 电穿孔 

分 类 号：R363[医药卫生—病理学]