Lipofectamine TM 2000介导pIRES2-EGFP-NT3转染小鼠耳蜗成纤维细胞的实验研究

摘要

目的：检测真核表达质粒载体pIRES2-EGFP-NT3在阳离子脂质体介导下对新生小鼠耳蜗成纤维细胞的转染情况。方法培养新生小鼠耳蜗成纤维细胞，用阳离子脂质体Lipofectamine TM 2000介导的方法，将含有目的基因NT3的真核表达质粒载体pIRES2-EGFP-NT3对其进行转染，转染后24 h，通过Confocal 显微镜观察新生小鼠耳蜗成纤维细胞的转染情况。结果通过阳离子脂质体Lipofectamine TM 2000介导，真核表达质粒载体pIRES2-EGFP-NT3能有效转染新生小鼠耳蜗成纤维细胞。结论含有目的基因NT3的真核表达质粒载体pIRES2-EGFP-NT3在阳离子脂质体介导下对新生小鼠耳蜗成纤维细胞的有效转染为研究NT3基因转染对正常及致聋小鼠耳蜗效应的实验奠定了基础。%Objective To detect the transfection of pIRES 2-EGFP-NT3 in mouse cochlea fibroblast by cationic liposome . Methods After pIRES2-EGFP-NT3 had been abstracted successfully , it was transfected into mouse cochlea fibroblast by lipofectami-neTM2000.Twenty four hours later, the efficiency of the transfection was analyzed by confocal microscope .Results The pIRES2-EG-FP-NT3 was effectively transfected into mouse cochlea fibroblast by cationic liposome .The transfected fibroblasts displaying green fluo-rescence were observed under fluorescence microscope .Conclusions The effective transfection of pIRES 2-EGFP-NT3 into mouse cochlea fibroblast by lipofectamine TM 2000 laid the basis for the following experiments , such as NT3 gene transfection in deaf or normal cochlea and so on .