Skeletal muscle tissue engineering: A maturation model promoting long-term survival of myotubes, structural development of the excitation-contraction coupling apparatus and neonatal myosin heavy chain expression

    Authors

    M. Das; J. W. Rumsey; N. Bhargava; M. Stancescu;J. J. Hickman

    Comments

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    Abbreviated Journal Title

    Biomaterials

    Keywords

    Cell culture; Silane; Muscle; In vitro test; Surface analysis; Surface; modification; SERUM-FREE MEDIUM; CILIARY NEUROTROPHIC FACTOR; LEUKEMIA INHIBITORY; FACTOR; ADULT HUMAN MYOBLASTS; SPINAL-CORD NEURONS; IN-VITRO; DEFINED; SYSTEM; DIHYDROPYRIDINE RECEPTOR; MYOGENIC DIFFERENTIATION; SILICON; MICROSTRUCTURES; Engineering, Biomedical; Materials Science, Biomaterials

    Abstract

    The use of defined in vitro systems to study the developmental and physiological characteristics of a variety of cell types is increasing, due in large part to their ease of integration with tissue engineering, regenerative medicine, and high-throughput screening applications. In this study, myotubes derived from fetal rat hind limbs were induced to develop several aspects of mature muscle including: sarcomere assembly, development of the excitation-contraction coupling apparatus and myosin heavy chain (MHC) class switching. Utilizing immunocytochemical analysis, anisotropic and isotropic band formation (striations) within the myotubes was established, indicative of sarcomere formation. In addition, clusters of ryanodine receptors were colocalized with dihydropyridine complex proteins which signaled development of the excitation-contraction coupling apparatus and transverse tubule biogenesis. The myotubes also exhibited MHC class switching from embryonic to neonatal MHC. Lastly, the myotubes survived significantly longer in culture (70-90 days) than myotubes from our previously developed system (20-25 days). These results were achieved by modifying the culture timeline as well as the development of a new medium formulation. This defined model system for skeletal muscle maturation supports the goal of developing physiologically relevant muscle constructs for use in tissue engineering and regenerative medicine as well as for high-throughput screening applications. (C) 2009 Elsevier Ltd. All rights reserved.

    Journal Title

    Biomaterials

    Volume

    30

    Issue/Number

    29

    Publication Date

    1-1-2009

    Document Type

    Article

    Language

    English

    First Page

    5392

    Last Page

    5402

    WOS Identifier

    WOS:000270115200016

    ISSN

    0142-9612

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