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Nonviral gene transfer into primary cultures of human and procine mesothelial cells

  • Biotechnology
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Summary

Due to their abundance and accessibility, mesothelial cells may be suitable tools for recombinant reagent expression by gene transfer. Genetically modified porcine mesothelial cells (PMCs) may have the potential for the treatment of vascular diseases in humans. We studied the effect of various transfection reagents on the primary culture of PMCs and human mesothelial cells (HMCs). The cells were transfected with a plasmid encoding a reporter gene (luciferase or green fluorescent protein [GFP]) under the control of the cytomegalovirus promoter. Transfection was achieved using cationic lipids (DOSPER and DOTAP) or calcium phosphate/deoxyribonucleic acid coprecipitation or Fugene 6. Results showed that Fugene 6 was the most efficient and reproducible transfection reagent with both PMCs and HMCs. With, Fugene 6, luciferase activity in PMCs (1.5×108 relative light units [RLU]/106 cells) was at least 2.5-fold higher than with the other transfection reagents, and it was 100-fold higher than in HMCs. However, the proportion of transfected cells expressing GFP was only 1%. These preliminary findings open up new avenues for developing experimental studies on the use of genetically modified PMCs.

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Authors and Affiliations

  1. Laboratoire de Chirurgie Vascularie et Theoracique, Paris, France

    J. Ohan & G. Leseche

  2. INSERM 353, Paris, France

    M. A. Gilbert

  3. INSERM 450 et Service d'Hématologie de l'hôpital Lariboisière, Paris, France

    L. Drouet

  4. Service de Chirurgie Générale de l'hôpital lariboisière, Paris, France

    Y. Panis

  5. Centre de Biophysique Moléculaire, Orléans, France

    P. Midoux

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  1. J. Ohan
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  2. M. A. Gilbert
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  3. G. Leseche
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  4. Y. Panis
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  5. P. Midoux
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  6. L. Drouet
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Ohan, J., Gilbert, M.A., Leseche, G. et al. Nonviral gene transfer into primary cultures of human and procine mesothelial cells. In Vitro Cell.Dev.Biol.-Animal 37, 402–407 (2001). https://doi.org/10.1290/1071-2690(2001)037<0402:NGTIPC>2.0.CO;2

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  • DOI: https://doi.org/10.1290/1071-2690(2001)037<0402:NGTIPC>2.0.CO;2

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