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Competitive electroporation formulation for cell therapy

Cancer Gene Therapy volume 18, pages 579–586 (2011)Cite this article

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Abstract

Established cell transfection via nucleofection relies on nucleofection buffers with unknown and proprietary makeup due to trade secrecy, inhibiting the possibility of using this otherwise effective method for developing cell therapy. We devised a three-step method for discovering an optimal formulation for the nucleofection of any cell line. These steps include the selection of the best nucleofection program and known buffer type, selection of the best polymer for boosting the transfection efficiency of the best buffer and the comparison with the optimal buffer from an established commercial vendor (Amaxa). Using this three-step selection system, competitive nucleofection formulations were discovered for multiple cell lines, which are equal to or surpass the efficiency of the Amaxa nucleofector solution in a variety of cells and cell lines, including primary adipose stem cells, muscle cells, tumor cells and immune cells. Through the use of scanning electron microscopy, we have revealed morphological changes, which predispose for the ability of these buffers to assist in transferring plasmid DNA into the nuclear space. Our formulation may greatly reduce the cost of electroporation study in laboratory and boosts the potential of application of electroporation-based cell therapies in clinical trials.

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Acknowledgements

This study was supported by National Institutes of Health grant RO1CA120895 and NIH/NIBIB grant R21EB007208. Authors also are thankful to the assistance in cell culture from Jiemiao Hu and in plasmid DNA preparation by Xueqing Xia.

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Authors and Affiliations

  1. Department of Comparative Biomedical Sciences, Louisiana State University, Baton Rouge, LA, USA

    M Flanagan, X Wu & S Yao

  2. Pennington Biomedical Research Center, Baton Rouge, LA, USA

    J M Gimble & G Yu

  3. Department of Pediatrics Research, University Texas MD Anderson Cancer Center, University Texas Graduate School of Biomedical Sciences, Holcombe, TX, USA

    X Xia, J Hu & S Li

Authors
  1. M Flanagan
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  2. J M Gimble
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  3. G Yu
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  7. S Yao
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  8. S Li
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Correspondence to S Li.

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Flanagan, M., Gimble, J., Yu, G. et al. Competitive electroporation formulation for cell therapy. Cancer Gene Ther 18, 579–586 (2011). https://doi.org/10.1038/cgt.2011.27

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