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Sci. STKE, 22 July 2003
Vol. 2003, Issue 192, p. pl11
[DOI: 10.1126/stke.2003.192.pl11]

PROTOCOLS

Biolistic Transfection of Cultured Myotubes

Christian Antolik1, Patrick G. De Deyne2, and Robert J. Bloch1*

1Departments of Physiology, University of Maryland School of Medicine, 655 W. Baltimore St., Baltimore, MD 21201 USA.
2Departments of Physical Therapy and Rehabilitation Sciences, University of Maryland School of Medicine, 655 W. Baltimore St., Baltimore, MD 21201 USA.

Abstract: Transfection of cells in culture with cDNA constructs is a powerful tool in cell biology, but postmitotic cells, including myotubes, can be hard to transfect with classic methods. Biolistics provides an alternative. We have used this biolistic technique to introduce cDNAs into cultured rat, chick, and C2C12 myotubes. This protocol results in efficient (20 to 70%, depending on cell type) transfection of myotubes, high levels of cDNA expression in individual myotubes, and little cellular damage. Using this procedure, we have expressed different muscle-specific cDNAs as green fluorescent protein (GFP) fusions. This technique is rapid, reliable, uses minimal amounts of reagent per transfection, and yields high transfection rates in a previously hard-to-transfect cell type. Its efficiency and reliability are high, regardless of plasmid size or epitope tag. Muscle cell biologists may now perform experiments in mature myotubes rather than relying on transfection of myoblast cultures or heterologous expression systems.

*Corresponding author. Telephone, 410-706-3020; e-mail, rbloch{at}umaryland.edu

Citation: C. Antolik, P. G. De Deyne, R. J. Bloch, Biolistic Transfection of Cultured Myotubes. Sci. STKE 2003, pl11 (2003).

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