Lipofection of cultured mouse muscle cells: a direct comparison of Lipofectamine and DOSPER
| Autor: | K Naujoks, George Dickson, Matthew G. Dunckley, E. Dodds, U Michaelis |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Green Fluorescent Proteins
Biology Transfection Viral vector Green fluorescent protein Mice Escherichia coli Genetics Animals Cation Exchange Resins Muscle Skeletal Molecular Biology Cells Cultured Reporter gene Histocytochemistry Genetic transfer Flow Cytometry beta-Galactosidase Lipids Molecular biology Luminescent Proteins Microscopy Electron Microscopy Fluorescence Cell culture Lipofectamine Molecular Medicine C2C12 |
| Zdroj: | Gene Therapy. 5:542-551 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3300604 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|