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

Due to their abundance and accessibility, mesothelial cells may be suitable toots for recombinant reagent expression by gene transfer. Genetically mod ified porcine mesothelial cells (PMCs) may have the potential for the treat ment of vascular diseases in humans. We studied the effect of various trans fection 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 c ytomegalovirus promoter. Transfection was achieved using cationic lipids (D OSPER and DOTAP) or calcium phosphate/deoxyribonucleic acid coprecipitation or Fugene 6. Results showed that Fugene 6 was the most efficient and repro ducible transfection reagent with both PMCs and HMCs. With Fugene 6, lucife rase activity in PMCs (1.5 X 10(8) relative light units [RLU]/10(6) 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 transfecte d cells expressing GFP was only 1%. These preliminary findings open up new avenues for developing experimental studies on the use of genetically modif ied PMCs.