Differentiation of skeletal muscle and integration of myotubes with silicon microstructures using serum-free medium and a synthetic silane substrate

    Authors

    M. Das; K. Wilson; P. Molnar;J. J. Hickman

    Comments

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    Abbreviated Journal Title

    Nat. Protoc.

    Keywords

    ORGANOSILANE SURFACE; DEFINED SYSTEM; IN-VITRO; RAT; NETWORKS; CULTURE; Biochemical Research Methods

    Abstract

    This protocol describes a cell culture model to study the differentiation of fetal rat skeletal muscle cells. The model uses serum-free medium, a nonbiological substrate N-1[3(trimethoxysilyl)propyl] diethylenetriamine ( DETA) and fabricated microcantilevers to promote the differentiation of dissociated rat myocytes into robust myotubes. In this protocol, we also describe how to characterize the myotubes on the basis of morphology, immunocytochemistry and electrophysiology. Here, four major techniques are employed: fabrication of cantilevers, surface modification of the glass and cantilever substrates with a DETA SAM, a serum-free medium and refined culture techniques. This culture system has potential applications in biocompatibility studies, bioartificial muscle engineering, skeletal muscle differentiation studies and for better understanding of myopathies and neuromuscular disorders. The model can be established in 26-33 d.

    Journal Title

    Nature Protocols

    Volume

    2

    Issue/Number

    7

    Publication Date

    1-1-2007

    Document Type

    Article

    Language

    English

    First Page

    1795

    Last Page

    1801

    WOS Identifier

    WOS:000253139200026

    ISSN

    1754-2189

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