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Efficiency of DNA Transfection of Rat Heart Myoblast Cells H9c2(2-1) by Either Polyethyleneimine or Electroporation

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Abstract

Expression of exogenous DNA in vitro is significantly affected by the particular transfection method utilized. In this study, we evaluated the efficiency of two transfection methods, chemically mediated polyethyleneimine (PEI) treatment and physically mediated electroporation, on a rat heart myoblast cell line, H9c2(2-1). After PEI transfection of pPgk-1/EGFP into H9c2(2-1) cells, EGFP expression could be easily detected by fluorospectrometer after 48 h (210 ± 12 RFU) and continued to increase after 72 h (243 ± 14 RFU). However, when H9c2(2-1) cells were transfected by electroporation (200 V, 500 μF, and one pulse), low level EGFP expression was observed after 48 h (49 ± 4 RFU) or 72 h (21 ± 14 RFU). In contrast, the easily transfectable control CHO-K1 cell line displayed a stronger EGFP expression than the H9c2(2-1) cells either by PEI or electroporation transfection. When transfection efficiencies were assayed by flow cytometry after 72 h, 13.6 ± 2.2% of PEI and 10.1 ± 1.5% of electroporation (250 V, 500 μF, and two pulses) transfected cells of H9c2(2-1) expressed EGFP, and PEI-transfected cells appeared to be less damaged (viability 93.6%) as compared to electroporation-transfected cells (39.5%). However, both PEI and electroporation (580 V, 50 Ω, and 50 μF) were effective for transfection of CHO-K1 with a higher efficiency, cell viability, and EGFP expression than H9c2(2-1). Our results indicate that the transfection efficiency of different methods varies among cell lines and that PEI is more efficient than electropolation for transfection of H9c2(2-1) whereas both PEI and electroporation are suitable for CHO-K1 transfection.

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Acknowledgments

We would like to thank reviewer and the editor Hyung Joon Cha, Ph.D. for their comments. The authors thank Dr. Maurice S. B. Ku for critical comments and revise of the manuscript. This work was supported by Nation Science Council, Taiwan (NSC-96-2313-B-415-007).

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Authors and Affiliations

  1. Department of Bioagricultural Science, National Chia Yi University, 300 Syuefu Rd., Chiayi, 60004, Taiwan

    Yu Chia Liu, Win Yu Lin & Hsi Tien Wu

  2. Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Ya Ru Jhang & Sing Hui Huang

  3. Department of Animal Science, National Chia Yi University, Chiayi, Taiwan

    Chean Ping Wu

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  1. Yu Chia Liu
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Correspondence to Hsi Tien Wu.

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Liu, Y.C., Lin, W.Y., Jhang, Y.R. et al. Efficiency of DNA Transfection of Rat Heart Myoblast Cells H9c2(2-1) by Either Polyethyleneimine or Electroporation. Appl Biochem Biotechnol 164, 1172–1182 (2011). https://doi.org/10.1007/s12010-011-9203-5

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