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A Comparison Study of Lipid Tranfection Reagents in A549, NIH 3T3 and COS-7,,,Cell Lines

>) that the highest transfection efficiency, as measured by b-galactosidase activity, occurred using 9 l of LipoTAXI reagent. This approach typically indicates the best amount of a transfection reagent for a particular cell line. Since high-level expression of a gene of interest may lead to unacceptably high cytotoxicity, total protein should be compared in cell lysates to select the optimal volume of LipoTAXI transfection reagent for the cell line studied.3

Figure 1

Comparison of Transfection Reagents

Using the results of the optimization experiment, we compared the efficacy of LipoTAXI transfection reagent to four other transfection reagents. Three of the reagents were cationic lipids; the fourth reagent was a branched polycationic polymer. All transfections included 1.0 g of reporter plasmid DNA. As determined for each cell line (Figure 1), the optimal amount of LipoTAXI transfection reagent was used. The other reagents were used at the volume recommended by the manufacturer. To ensure a fair comparison study, we also used the other reagents at slightly higher and lower amounts than recommended.

Figure 2

Analysis

The optimal amount of LipoTAXI transfection reagent varied considerably between cell lines. In general, the amount of transfection reagent that can be used is limited by cytotoxic effects of either the lipid or the expressed plasmid protein. The greater the amount of lipid tolerated by the cell
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