Improved lipid-mediated gene transfer in C6 glioma cells and primary glialcells using FuGene((TM))

Gene therapy is a potent method to counteract neurodegeneration by introduc ing genetic information encoding neuroprotective factors. In this study cat ionic lipids were used to transfer DNA into C6 glioma cells and primary gli al cells. When comparing the novel compound FuGene with other commercially- available lipids, it was found that FuGene markedly enhanced gene transfer of a beta-galactosidase reporter plasmid into C6 glioma cells. FuGene had s everal advantages compared to other lipids, such as a very low toxicity and the capability of transfection under serum conditions. When optimizing, a DNA-lipid ratio of 150 ng DNA/1 mu l FuGene and a concentration of 3 mu l F uGene/1 mi medium was found to be optimal. The incubation time peaked after 8 h and the expression time reached an optimum between 2 and 6 days. When cells were transfected on 3 consecutive days for 6 h each ('boosting'), the transfection efficiency was markedly enhanced in primary glial cells. When using endotoxin-free DNA the transfection efficiency could be enhanced up to 3 times. The optimal transfection efficiency in C6 glioma cells and in p rimary glial cells was found to be 16.3 +/- 0.3% and 5.1 +/- 0.37% of total cells, respectively. In conclusion this study shows that the novel compoun d FuGene has a very high potential to transfer DNA into cells of glial orig in, and it might be an interesting canditate for ex vivo and in vivo gene t herapeutic approaches. (C) 1999 Elsevier Science B.V. All rights reserved.