Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze Satellite Cells View Full Text


Ontology type: schema:Chapter      Open Access: True


Chapter Info

DATE

2004-10-15

AUTHORS

Cheryl D. Helgason , Cindy Miller , Gabi Shefer , Zipora Yablonka-Reuveni

ABSTRACT

Myofibers are the functional contractile units of skeletal muscle. Mononuclear satellite cells located between the basal lamina and the plasmalemma of the myofiber are the primary source of myogenic precursor cells in postnatal muscle. This chapter describes protocols used in our laboratory for isolation, culturing, and immunostaining of single myofibers from mouse skeletal muscle. The isolated myofibers are intact and retain their associated satellite cells underneath the basal lamina. The first protocol discusses myofiber isolation from the flexor digitorum brevis (FDB) muscle. Myofibers are cultured in dishes coated with Vitrogen collagen, and satellite cells remain associated with the myofibers undergoing proliferation and differentiation on the myofiber surface. The second protocol discusses the isolation of longer myofibers from the extensor digitorum longus (EDL). Different from the FDB myofibers, the longer EDL myofibers tend to tangle and break when cultured together; therefore, EDL myofibers are cultured individually. These myofibers are cultured in dishes coated with Matrigel. The satellite cells initially remain associated with the myofiber and later migrate away to its vicinity, resulting in extensive cell proliferation and differentiation. These protocols allow studies on the interplay between the myofiber and its associated satellite cells. More... »

PAGES

281-304

References to SciGraph publications

  • 1995-11. Culturing satellite cells from living single muscle fiber explants in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - ANIMAL
  • Book

    TITLE

    Basic Cell Culture Protocols

    ISBN

    1-59259-838-2

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1385/1-59259-838-2:281

    DOI

    http://dx.doi.org/10.1385/1-59259-838-2:281

    DIMENSIONS

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