Abstract
The technique of single muscle-fiber cultures has already proven valuable in extending knowledge of myogenesis, stem cell heterogeneity, the stem cell niche in skeletal muscle, and satellite cell activation. This report reviews the background of the model and applications, and details the procedures of muscle dissection, fiber digestion and isolation, cleaning the fiber preparation, plating fibers, and extensions of the technique for studying activation from stable quiescence of satellite cells, mRNA expression by in situ hybridization and regulation of satellite cell activation in zebrafish muscle by nitric oxide, hepatocyte growth factor.
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Acknowledgments
The work of a dedicated research technician (Ritika Upadhaya), summer research students (Alyssa Janke, Stéphane Lenoski, Melody Ong, Jacqueline Richelle, and Colin Rumbolt) and a graduate student (Dr. Orest Pilipowicz, MSc, DMD) provided experiments illustrating effects of drug screening on mouse fibers (SL & CR, see Fig. 1), marcaine in mouse fibers (OP, see Fig. 3) and ISDN and HGF treatments in zebrafish fiber cultures (AJ, MO and JR, see Fig. 4). Funding for this work (to JEA) was from the Canadian Space Agency, the Manitoba Institute of Child Health (MICH), and the Natural Sciences and Engineering Research Council (NSERC). Student support was received from NSERC UnderÂgraduate Summer Research Scholarships (AJ, MO, CR, JR), Faculty of Science University of Manitoba Undergraduate Scholarships (SL), a MICH Graduate Scholarship (OP), a Canada Graduate Scholarship (ACW) and a post-doctoral scholarship from the Canadian Bureau for International Education (WM).
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Anderson, J.E., Wozniak, A.C., Mizunoya, W. (2012). Single Muscle-Fiber Isolation and Culture for Cellular, Molecular, Pharmacological, and Evolutionary Studies. In: DiMario, J. (eds) Myogenesis. Methods in Molecular Biology, vol 798. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-343-1_6
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DOI: https://doi.org/10.1007/978-1-61779-343-1_6
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