不同基因转染方法对小鼠内耳毛细胞转染效率的比较

The Comparative Study of Different Methods for Gene Transfer to Cochlear Hair Cells

陈岩;赖垂金;李雯;李华伟;

1:复旦大学附属眼耳鼻喉科医院

摘要
目的对比不同的基因转染方法对小鼠内耳毛细胞的转染效率,以探寻简便、高效转染耳蜗毛细胞的方法。方法分离111只新生昆明小鼠(P2)耳蜗螺旋韧带及感染上皮,分别采用电穿孔法介导质粒载体(pGPHI/GFP/Neo)、腺病毒载体(Ad5/CMV/GFP)和慢病毒载体(LV/CMV/GFP)3种方法转染小鼠内耳毛细胞,于转染48小时后在荧光显微镜下观察并比较三种方法基因转染后小鼠内耳毛细胞绿色荧光蛋白(green fluorescence protein,GFP)的表达。结果电穿孔法介导质粒载体转染组和慢病毒载体转染组GFP阳性细胞极少;腺病毒载体转染组效率最高,耳蜗中回外毛细胞GFP阳性率为90.0%±4.1%,内毛细胞GFP阳性率为5%±0.4%。结论腺病毒载体能更有效地将外源基因导入耳蜗毛细胞内表达,是基因转染耳蜗毛细胞的理想载体。
关键词
腺病毒载体;慢病毒载体;电穿孔法;基因转染;内耳毛细胞
基金项目(Foundation):
973及国家重大研究计划(2011CB504506);; 国家自然科学基金(81100709);; 教育部博士点基金博导类计划(20120071110077)联合资助
作者
陈岩;赖垂金;李雯;李华伟;
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