Characterization and optimization of a simple, repeatable system for the long term in vitro culture of aligned myotubes in 3D

Authors

  • A.S.T. Smith,

    1. The Sobell Department of Motor Neuroscience and Movement Disorders, UCL Institute of Neurology, London, UK
    2. Muscle Cellular and Molecular Physiology Research Group, Institute of Sport and Physical Activity Research (ISPAR Bedford), University of Bedfordshire, Bedford, UK
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  • S. Passey,

    1. Muscle Cellular and Molecular Physiology Research Group, Institute of Sport and Physical Activity Research (ISPAR Bedford), University of Bedfordshire, Bedford, UK
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  • L. Greensmith,

    1. The Sobell Department of Motor Neuroscience and Movement Disorders, UCL Institute of Neurology, London, UK
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  • V. Mudera,

    1. UCL Institute of Orthopaedics and Musculoskeletal Science, London, UK
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  • M.P. Lewis

    Corresponding author
    1. Muscle Cellular and Molecular Physiology Research Group, Institute of Sport and Physical Activity Research (ISPAR Bedford), University of Bedfordshire, Bedford, UK
    2. UCL School of Life and Medical Sciences, London, UK
    3. Loughborough University, School of Sport, Health and Exercise Sciences, Loughborough, UK
    4. Cranfield University, Cranfield Health, Cranfield, UK
    • Muscle Cellular and Molecular Physiology Research Group, Institute of Sport and Physical Activity Research, Bedford, Polhill Avenue, Bedford MK41 9EA, London, UK.
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  • Work performed at UCL Institute of Neurology and the University of Bedfordshire.

Abstract

Increased recent research activity in exercise physiology has dramatically improved our understanding of skeletal muscle development and physiology in both health and disease. Advances in bioengineering have enabled the development of biomimetic 3D in vitro models of skeletal muscle which have the potential to further advance our understanding of the fundamental processes that underpin muscle physiology. As the principle structural protein of the extracellular matrix, collagen-based matrices are popular tools for the creation of such 3D models but the custom nature of many reported systems has precluded their more widespread adoption. Here we present a simple, reproducible iteration of an established 3D in vitro model of skeletal muscle, demonstrating both the high levels of reproducibility possible in this system and the improved cellular architecture of such constructs over standard 2D cell culture techniques. We have used primary rat muscle cells to validate this simple model and generate comparable data to conventional established cell culture techniques. We have optimized culture parameters for these cells which should provide a template in this 3D system for using muscle cells derived from other donor species and cell lines. J. Cell. Biochem. 113: 1044–1053, 2012. © 2011 Wiley Periodicals, Inc.

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