胶质瘤U251细胞外源基因瞬时转染方法的选择

发布时间：2019-08-07 16:45

【摘要】：目的选择一种将外源基因瞬时转入胶质瘤细胞的高效转染方法。方法以去内毒素真核表达载体DsRed2为外源基因,分别采用脂质体Lipofectamine2000转染法、脂质体Viafect转染法以及电转染法,将不同浓度的DsRed2质粒(0.25、0.5、1.0μg)转染入胶质瘤U251细胞(以下称胶质瘤细胞,细胞数量均为1.0×105个)。荧光显微镜下采用可见光、红色荧光滤镜观察细胞形态及红色荧光蛋白表达,紫外光滤镜下观察细胞核Hocest33342染色情况,计算转染效率。结果三种方法均能将质粒转染至胶质瘤细胞中,红色荧光蛋白表达效果均良好。转染后的细胞形态基本正常,但表达红色荧光蛋白与未表达红色荧光蛋白的细胞形态差异较大。相同质粒浓度下,电转染法转染后表达红色荧光蛋白的细胞数量多于脂质体Lipofectamine2000转染法、脂质体Viafect转染法。随着质粒浓度的升高,脂质体Viafect转染法和电转染法对胶质瘤细胞的转染效率逐渐升高。相同质粒浓度下,电转染法对胶质瘤细胞的转染效率均高于脂质体Lipofectamine2000转染法和脂质体Viafect转染法(P均0.01)。结论电转染法对外源基因瞬时转染胶质瘤细胞的转染效率较高,并呈质粒浓度依赖性。
[Abstract]:Objective to select an efficient method for transient transfer of foreign genes into glioma cells. Methods Eukaryotic expression vector DsRed2 was used as foreign gene to transfer different concentrations of DsRed2 plasmid (0.25 渭 g, 0.5 渭 g, 1.0 渭 g) into glioma U251 cells (hereinafter referred to as glioma cells, the number of cells was 1.0 脳 10 5) by liposomes Lipofectamine2000, Viafect and electroporation, respectively. The morphology of cells and the expression of red fluorescent protein were observed by visible light and red fluorescence filter under fluorescence microscope, and the staining of nuclear Hocest33342 was observed under ultraviolet light filter, and the transfer efficiency was calculated. Results all the three methods could transfer the plasmid into glioma cells, and the expression of red fluorescent protein was good. The morphology of the cells after transfection was basically normal, but the morphology of the cells expressing red fluorescent protein was different from that of non-expressing red fluorescent protein. At the same plasmid concentration, the number of cells expressing red fluorescent protein after electrostaining was more than that of liposomes Lipofectamine2000 and Viafect. With the increase of plasmid concentration, the efficiency of liposomes Viafect and electroporation on glioma cells increased gradually. At the same plasmid concentration, the efficiency of electroporation on glioma cells was higher than that of liposomes Lipofectamine2000 and Viafect. Conclusion the efficiency of transient transfer of foreign genes into glioma cells is high in a concentration-dependent manner.
【作者单位】：内蒙古医科大学附属医院;
【基金】：内蒙古医科大学科技百万工程(YKD2015KJBW010);内蒙古医科大学青年创新基金资助项目(YKD2012QNX009)
【分类号】：R739.41

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