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Optimization of a gene electrotransfer method for mesenchymal stem cell transfection

Gene Therapy volume 15, pages 537–544 (2008)Cite this article

Abstract

Gene electrotransfer is an efficient and reproducible nonviral gene transfer technique useful for the nonpermanent expression of therapeutic transgenes. The present study established optimal conditions for the electrotransfer of reporter genes into mesenchymal stem cells (MSCs) isolated from rat bone marrow by their selective adherence to tissue-culture plasticware. The electrotransfer of the lacZ reporter gene was optimized by adjusting the pulse electric field intensity, electric pulse type, electropulsation buffer conductivity and electroporation temperature. LacZ electrotransfection into MSCs was optimal at 1500 V cm−1 with pre-incubation in Spinner's minimum essential medium buffer at 22 °C. Under these conditions β-galactosidase expression was achieved in 29±3% of adherent cells 48 h post transfection. The kinetics of β-galactosidase activity revealed maintenance of β-galactosidase production for at least 10 days. Moreover, electroporation did not affect the MSC potential for multidifferentiation; electroporated MSCs differentiated into osteoblastic, adipogenic and chondrogenic lineages to the same extent as cells that were not exposed to electric pulses. Thus, this study demonstrates the feasibility of efficient transgene electrotransfer into MSCs while preserving cell viability and multipotency.

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Acknowledgements

We thank Ceraver Company (Gonesse, France) and the Fondation de l' Avenir (Grant ET7-479) for the financial support. We also thank K Oudina for performing statistical analyses and express special thanks to R Bizios for critical reading of the manuscript.

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Authors and Affiliations

  1. Laboratoire de Recherches Orthopédiques (B2OA), CNRS UMR 7052, Paris, France

    E Ferreira, E Potier, D Logeart-Avramoglou & H Petite

  2. AO Research Institute, Davos, Switzerland

    E Potier

  3. CNRS UMR 8121, Institut Gustave-Roussy, Villejuif, France

    S Salomskaite-Davalgiene & L M Mir

  4. Biology Department, Biophysical Research Laboratory, Vytautas Magnus University, Kaunas, Lithuania

    S Salomskaite-Davalgiene

  5. Univ Paris-Sud, UMR 8121, Villejuif, France

    L M Mir

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  1. E Ferreira
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Correspondence to H Petite.

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Ferreira, E., Potier, E., Logeart-Avramoglou, D. et al. Optimization of a gene electrotransfer method for mesenchymal stem cell transfection. Gene Ther 15, 537–544 (2008). https://doi.org/10.1038/gt.2008.9

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