Optimization of Electrotransfection Conditions of Mammalian Cells with Different Biological Features
Optimization of Electrotransfection Conditions of Mammalian Cells with Different Biological Features
复制标题
不同生物学特性哺乳动物细胞电转染条件的优化
DOI：
10.1007/s00232-012-9480-0
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发表时间：
2012-07
影响因子：
2.4
通讯作者：
Zhang, Zhencang
中科院分区：
文献类型：
--
作者：
Guo, Huichen;Hao, Rongzeng;Wei, Yanquan;Sun, Dehui;Sun, Shiqi;Zhang, Zhencang
采用电穿孔法将编码绿色荧光蛋白（GFP）基因的真核表达质粒pEGFP-N1导入不同生物学特性的细胞中。通过荧光显微镜观察和台盼蓝染色观察转染后细胞存活率，研究电压、电容流量、脉冲周期、DNA用量和缓冲液等条件对转染效率的影响。在以下上皮细胞中实现了更好的电转染结果：300 V/850 μF的Vero细胞、300 V/500 μF的PK 15细胞、200 V/600 μF的MDCK细胞、200 V/500 μF的F81细胞、300 V/400 μ F的癌细胞MB 49、200 V/450 μF的Hela细胞，HF-29电池为300 V/800 μF，B16 F1电池为200 V/650 μF。在成纤维细胞中，BHK 21细胞在300 V/600 μF下和ST细胞在200 V/750 μF下实现了更好的电转染。不含抗生素和血清的RPMI-1640培养基作为电穿孔缓冲液，电转染效率和细胞存活率均高于其他细胞培养基。我们的研究结果进一步证明电穿孔转染是一种有效的基因转染方法。不同生物学特性的细胞需要不同的转染条件以获得更高的目的基因转染效率。
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 μF, PK15 cells at 300 V/500 μF, MDCK cells at 200 V/600 μF, F81 cells at 200 V/500 μF, cancer cells MB49 at 300 V/400 μF, Hela cells at 200 V/450 μF, HF-29 cells at 300 V/800 μF and B16F1 cells at 200 V/650 μF. Among fibroblast cells, better electrotransfection was achieved in BHK21 cells at 300 V/600 μF and ST cells at 200 V/750 μF. 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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影响因子：
--
作者：
R. Kingston;Claudia A. Chen;H. Okayama
通讯作者：
R. Kingston;Claudia A. Chen;H. Okayama
影响因子：
3.4
作者：
Canatella, PJ;Karr, JF;Prausnitz, MR
通讯作者：
Prausnitz, MR
影响因子：
2.7
作者：
C. Baum;P. Forster;S. Hegewisch-Becker;K. Harbers
通讯作者：
C. Baum;P. Forster;S. Hegewisch-Becker;K. Harbers
影响因子：
--
作者：
H. Robinson;T. Pertmer
通讯作者：
H. Robinson;T. Pertmer
影响因子：
5
作者：
Cegovnik, U;Novakovic, S
通讯作者：
Novakovic, S