Optimization of Electrotransfection Conditions of Mammalian Cells with Different Biological Features

文献类型： 外文期刊

第一作者： Guo, Huichen

作者： Guo, Huichen;Hao, Rongzeng;Wei, Yanquan;Sun, Dehui;Sun, Shiqi;Guo, Huichen;Hao, Rongzeng;Wei, Yanquan;Sun, Dehui;Sun, Shiqi;Zhang, Zhencang

作者机构：

关键词： Electroporation transfection;Mammalian cell;Parameter optimization

期刊名称：JOURNAL OF MEMBRANE BIOLOGY （ 影响因子：1.843； 五年影响因子：2.053 ）

ISSN： 0022-2631

年卷期： 2012 年 245 卷 12 期

页码：

收录情况： SCI

摘要： We introduced eukaryotic expression plasmid pEGFP-N1 encoding green fluorescent protein (GFP) genes into cells with different biological features through electroporation. The effects of conditions, including voltage, capacitor flow, pulse cycle, DNA dosage and buffer, on transfection efficiency were investigated based on fluorescent microscopy and posttransfection survival rate of cells by staining with trypan blue. Better electrotransfection outcomes were achieved in the following epithelial cells: Vero cells at 300 V/850 mu F, PK15 cells at 300 V/500 mu F, MDCK cells at 200 V/600 mu F, F81 cells at 200 V/500 mu F, cancer cells MB49 at 300 V/400 mu F, Hela cells at 200 V/450 mu F, HF-29 cells at 300 V/800 mu F and B16F1 cells at 200 V/650 mu F. Among fibroblast cells, better electrotransfection was achieved in BHK21 cells at 300 V/600 mu F and ST cells at 200 V/750 mu F. RPMI-1640 medium without antibiotics and serum demonstrated higher electrotransfection efficiency and cell survival rate than other cell culture media as electroporation buffer. Our findings further prove that electroporation transfection is an effective method for genetic transfection. Cells with different biological features require varying transfection conditions to obtain higher transfection efficiency of target genes.

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