脂质体介导外源基因体外转染兔角膜内皮细胞条件的优化  

Optimization of parameters of exogene transfection of rabbit corneal endothelial cell in vitro mediated by liposome

作　　者：毛晓春[1] 李贵刚[2] 张虹[2] 

机构地区：[1]华中科技大学同济医学院附属襄樊医院眼科,中国湖北省襄樊市441021 [2]华中科技大学同济医学院附属同济医院眼科,中国湖北省武汉市430030

出　　处：《国际眼科杂志》2008年第7期1353-1355,共3页International Eye Science

基　　金：中国湖北省自然科学基金资助项目(No.2003ABA150)~~

摘　　要：目的：观察阳离子脂质体LipofectamineTM 2000是否能介导目的基因转移至兔角膜内皮细胞,并优化转染条件。方法：体外培养兔眼角膜内皮细胞,以绿色荧光蛋白（GFP）为报告基因,通过体外细胞转染实验,用不同的细胞融合度、脂质体浓度、脂质体与质粒的比例、转染时间优化转染条件,荧光显微镜观察目的基因的表达。结果：角膜内皮细胞内可见脂质体介导的绿色荧光蛋白表达,LipofectamineTM在细胞融合度为80%～90%,脂质体/DNA为3L/g,脂质体的量为1.8μL,转染时间为5h时,转染效率最高。结论：阳离子脂质体可有效介导目的基因转移至角膜内皮细胞内,GFP可作为报告基因优化脂质体转染条件。AIM： To evaluate the efficiency of liposome mediated gene transfer to corneal endothelial. METHODS： Rabbit endothelium cells were cultured for in vitro test. With green fluorescence protein （GFP） as report gene, the transfection condition was optimized in cell confluence, liposome quantity, liposome/DNA ratio and transfecting time. RESULTS： The GFP expression in corneal endothelial cells was observed. The highest transfecting efficiency of LipofectAMINE2000 would be obtained when cell confluence was 80%-90%, liposome concentration 1.8μ L, ratio of liposome/DNA3L/g and transfecting time was 5 hours. CONCLUSION： Liposome is able to deliver genes to corneal endothelial cells with efficacy, the experiment impled result show that GFP is a convenient reporter gene.

关 键 词：角膜 内皮细胞 脂质体 基因转染 

分 类 号：R346[医药卫生—基础医学]