Gene Therapy
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April 1998, Volume 5, Number 4, Pages 542-551
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Paper
Lipofection of cultured mouse muscle cells: a direct comparison of Lipofectamine and DOSPER
E Dodds1, M G Dunckley1, K Naujoks2, U Michaelis2 and G Dickson1

1Division of Biochemistry, School of Biological Sciences, Royal Holloway University of London, Egham, Surrey TW20 0EX, UK

2Division of Therapeutics, Boehringer Mannheim GmbH, Penzberg, Germany

Abstract

Cationic lipid-DNA complexes (lipoplexes) have been widely used as gene transfer vectors which avoid the adverse immunogenicity and potential for viraemia of viral vectors. With the long-term aim of gene transfer into skeletal muscle in vivo, we describe a direct in vitro comparison of two commercially available cationic lipid formulations, Lipofectamine and DOSPER. Optimisation of transfection was performed in the C2C12 mouse muscle cell line, before further studies in primary mouse myoblasts and C2C12 myotubes. Reporter gene constructs expressing either E. coli beta-galactosidase or green fluorescent protein (GFP) were used in order to evaluate transfection efficiency by histochemical staining 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, DOSPER exhibited the important advantage of being able to transfect cells in the presence of serum of both bovine and murine origin. This feature allowed increased cell survival during in vitro transfections, and may be advantageous for direct in vivo gene transfer efficacy.

Keywords

gene transfer; lipoplex; lipofection; skeletal muscle

Received 8 August 1997; accepted 17 November 1997
April 1998, Volume 5, Number 4, Pages 542-551
Table of contents    Previous  Abstract  Next   Article  PDF