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Optimization of Electrotransfection Conditions of Mammalian Cells with Different Biological Features

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Abstract

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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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grants 31100688 and 31101838), the Foundation of Science and Technology Committee of Gansu (Grants 1102NKDA034 and 1102NKDA033) and the International Science and Technology Cooperation Program of Gansu (Grant 1104WCGA185).

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Authors and Affiliations

  1. State Key Laboratory of Veterinary Etiological Biology and National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, People’s Republic of China

    Huichen Guo, Rongzeng Hao, Yanquan Wei, Dehui Sun & Shiqi Sun

  2. Yangling Vocational and Technical College, Yangling, 712100, Shanxi, People’s Republic of China

    Zhencang Zhang

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  1. Huichen Guo
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  2. Rongzeng Hao
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  3. Yanquan Wei
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  4. Dehui Sun
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  5. Shiqi Sun
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  6. Zhencang Zhang
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Corresponding author

Correspondence to Shiqi Sun.

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H. Guo and R. Hao contributed equally to this work.

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Guo, H., Hao, R., Wei, Y. et al. Optimization of Electrotransfection Conditions of Mammalian Cells with Different Biological Features. J Membrane Biol 245, 789–795 (2012). https://doi.org/10.1007/s00232-012-9480-0

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  • DOI: https://doi.org/10.1007/s00232-012-9480-0

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