LIPOFECTION OF CULTURED MOUSE MUSCLE-CELLS - A DIRECT COMPARISON OF LIPOFECTAMINE AND DOSPER

Cationic lipid-DNA complexes (lipoplexes) have been widely used as gen e transfer vectors which avoid the adverse immunogenicity and potentia l for viraemia of viral vectors. With the long-term aim of gene transf er into skeletal muscle in vivo, we describe a direct in vitro compari son of two commercially available cationic lipid formulations, Lipofec tamine and DOSPER. Optimisation of transfection was performed in the C 2C12 mouse muscle cell line, before further studies in primary mouse m yoblasts and C2C12 myotubes. Reporter gene constructs expressing eithe r E. coli beta-galactosidase or green fluorescent protein (GFP) were u sed in order to evaluate transfection efficiency by histochemical stai ning or FACS analysis, respectively. Both lipid formulations were able to promote efficient, reproducible gene transfer in C2C12 cells, and to transfect primary mouse myoblast cultures successfully. However, DO SPER exhibited the important advantage of being able to transfect cell s in the presence of serum of both bovine and murine origin. This feat ure allowed increased cell survival during in vitro transfections, and may be advantageous for direct in vivo gene transfer efficacy.