Abstract
Muscle tissue culture provides a system for studying the growth and differentiation of muscle cells in a controlled environment. In mature muscle tissue, terminally differentiated myocytes form multinucleate syncytia in which structural and regulatory genes are expressed and the contractile apparatus is assembled. Adult muscle fibres are characterized by the presence of satellite cells. These are a quiescent population of myogenic cells that reside between the basal lamina and the plasmalemma of terminally differentiated muscle fibres and are rapidly activated in response to appropriate stimuli. This chapter describes protocols used in our laboratory for isolating and culturing satellite cells isolated from mouse skeletal muscles. In particular we discuss the technical aspect of satellite cell isolation, the methods necessary to enrich the satellite cell fraction, and the culture conditions which optimize proliferation and myotube formation of mouse satellite cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cossu, G., De Angelis, L., Borello, U., Berarducci, B., Buffa, V., Sonnino, C., Coletta, M. et al. (2000) Determination, diversification and multipotency of mammalian myogenic cells. Int. J. Dev. Biol. 44, 699–706.
Edmondson, D. G., and Olson, E. N. (1993) Helix-loop-helix proteins as regulators of muscle-specific transcription. J. Biol. Chem. 268, 755–758.
Sabourin, L. A., and Rudnicki, M. A. (2000) The molecular regulation of myogenesis. Clin. Genet. 57, 16–25.
Buckingham, M. (1992) Making muscle in mammals. Trends Genet. 8, 144–148.
Katz, B. (1961) The terminations of the afferent nerve fibre in the muscle spindle of the frog. Phil. Transact. Royal Soc. London Ser. B. 243, 221–240.
Mauro, A. (1961) Satellite cell of skeletal muscle fibers. J. Biophys. Biochem. Cytol. 9, 493–495.
Hawke, T. J., and Garry, D. J. (2001) Myogenic satellite cells: physiology to molecular biology. J. Appl. Physiol. 91, 534–551.
Buckingham, M., Bajard, L., Chang, T., Daubas, P., Hadchouel, J., Meilhac, S., Montarras, D., Rocancourt, D., and Relaix, F. (2003) The formation of skeletal muscle: from somite to limb. J. Anat. 202, 59–68.
Yan, Z., Choi, S., Liu, X., Zhang, M., Schageman, J. J., Lee, S. Y., Hart, R., Lin, L., Thurmond, F. A., and Williams, R. S. (2003) Highly coordinated gene regulation in mouse skeletal muscle regeneration. J. Biol. Chem. 278, 8826–8836.
Charge, S. B., and Rudnicki, M. A. (2004) Cellular and molecular regulation of muscle regeneration. Physiol. Rev. 84, 209–238.
Shefer, G., and Yablonka-Reuveni, Z. (2005) Isolation and culture of skeletal muscle myofibers as a means to analyze satellite cells. Methods Mol. Biol. 290, 281–304.
Collins, C. A., Olsen, I., Zammit, P. S., Heslop, L., Petrie, A., Partridge, T. A., and Morgan, J. E. (2005) Stem cell function, self-renewal, and behavioural heterogeneity of cells from the adult muscle satellite cell niche. Cell 122, 289–301.
Collins, C. A., and Zammit, P. S. (2009) Isolation and grafting of single muscle fibres. Methods Mol. Biol. 482, 319–330.
Montarras, D., Morgan, J., Collins, C., Relaix, F., Zaffran, S., Cumano, A., Partridge, T., and Buckingham, M. (2005) Direct isolation of satellite cells for skeletal muscle regeneration. Science 309, 2064–2067.
Acknowledgements
Work in the authors’ laboratory has been supported by Telethon, AIRC, ASI, Myoage-European network, and Fondazione Roma.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Musarò, A., Barberi, L. (2010). Isolation and Culture of Mouse Satellite Cells. In: Ward, A., Tosh, D. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 633. Humana Press. https://doi.org/10.1007/978-1-59745-019-5_8
Download citation
DOI: https://doi.org/10.1007/978-1-59745-019-5_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-772-3
Online ISBN: 978-1-59745-019-5
eBook Packages: Springer Protocols