Efficient lipid-mediated transfection of DNA into primary XXXX hepatocytes.

@entity11505 @entity14942 @entity23210 @entity35 @entity53536 @entity23209
Cationic lipids are an effective means for transfecting nucleic acids into a variety of cell types. Very few of these lipids, however, have been reported to be effective with primary cells. We report on the efficacy of several commercially available cationic lipid reagents to transfect plasmid DNA into primary @entity35 hepatocytes in culture. The reagents tested in this study include TransfectAce, LipofectAmine, Lipofectin, @entity53536 ( @entity53536 ), ( @entity11505 ( @entity11505 ), and @entity14942 / @entity23209 ( @entity14942 / @entity23210 ). Electron micrographic (EM) studies indicate that similar size Lipofectin and @entity11505 vesicles contain DNA-like material internally and that these vesicles attach to the cell membrane. @entity11505 vesicles are multilamellar, appear as clusters, and have a high DNA-to-lipid ratio. Lipofectin vesicles appear to attach to the cell surface as individual vesicles. The EM observations are consistent with current theories on the mechanism of transfection by cationic lipids. While Lipofectin has proven to be effective in transfection studies of primary cells in culture, we have found @entity11505 to be a viable alternative. @entity11505 yields transfection rates in hepatocytes comparable to @entity53536 and Lipofectin, however, at lower concentrations of reagent and at considerably less cost. Optimal conditions for transfecting 5 micrograms of plasmid DNA with @entity11505 were achieved by utilizing multilamellar (vortexed) vesicles at a concentration of 15 micrograms @entity11505 per 2 ml media in 60-mm plates for 2 h transfection time. In this study, @entity11505 has proven to be economical, easy to prepare, and very effective in transfecting DNA into primary @entity35 hepatocytes.
