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 β-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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dodds, E., Dunckley, M., Naujoks, K. et al. Lipofection of cultured mouse muscle cells: a direct comparison of Lipofectamine and DOSPER. Gene Ther 5, 542–551 (1998). https://doi.org/10.1038/sj.gt.3300604
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3300604
Keywords
This article is cited by
-
Virus-free transfection, transient expression, and purification of human cardiac myosin in mammalian muscle cells for biochemical and biophysical assays
Scientific Reports (2023)
-
Improved CRISPR/Cas9 gene editing in primary human myoblasts using low confluency cultures on Matrigel
Skeletal Muscle (2021)
-
Comparison of Transfection Efficiency of Nonviral Gene Transfer Reagents
Molecular Biotechnology (2010)
-
Analysis of Novel Nonviral Gene Transfer Systems for Gene Delivery to Cells of the Musculoskeletal System
Molecular Biotechnology (2008)
-
Study on biodistribution and imaging of radioiodinated antisense oligonucleotides in nude mice bearing human lymphoma
Journal of Radioanalytical and Nuclear Chemistry (2007)