Get access

Reduction of myoblast differentiation following multiple population doublings in mouse C2C12 cells: A model to investigate ageing?

Authors

  • Adam P. Sharples,

    Corresponding author
    1. Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health (IRM), Manchester Metropolitan University, John Dalton Building, Oxford Road, Manchester, UK
    2. Muscle Cellular and Molecular Physiology Research Group (MCMPRG), Department of Sport and Exercise Sciences, Institute for Sport and Physical Activity Research (ISPAR Bedford), University of Bedfordshire, London, UK
    • Muscle Cellular and Molecular Physiology Research Group (MCMPRG), Department of Sport and Exercise Sciences, Institute for Sport and Physical Activity Research (ISPAR Bedford), University of Bedfordshire, Polhill Avenue, Bedford, MK41 9EA, UK.
    Search for more papers by this author
  • Nasser Al-Shanti,

    1. Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health (IRM), Manchester Metropolitan University, John Dalton Building, Oxford Road, Manchester, UK
    Search for more papers by this author
  • Mark P. Lewis,

    1. Muscle Cellular and Molecular Physiology Research Group (MCMPRG), Department of Sport and Exercise Sciences, Institute for Sport and Physical Activity Research (ISPAR Bedford), University of Bedfordshire, London, UK
    2. School of Life and Medical Sciences, University College London (UCL), London, UK
    3. School of Sport, Health and Exercise Science, Loughborough University, Loughborough, UK
    4. Cranfield Health, Cranfield University, Cranfield, Bedfordshire, UK
    Search for more papers by this author
  • Claire E. Stewart

    1. Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health (IRM), Manchester Metropolitan University, John Dalton Building, Oxford Road, Manchester, UK
    Search for more papers by this author

Abstract

Ageing skeletal muscle displays declines in size, strength, and functional capacity. Given the acknowledged role that the systemic environment plays in reduced regeneration (Conboy et al. [2005] Nature 433: 760–764), the role of resident satellite cells (termed myoblasts upon activation) is relatively dismissed, where, multiple cellular divisions in-vivo throughout the lifespan could also impact on muscular deterioration. Using a model of multiple population doublings (MPD) in-vitro thus provided a system in which to investigate the direct impact of extensive cell duplications on muscle cell behavior. C2C12 mouse skeletal myoblasts (CON) were used fresh or following 58 population doublings (MPD). As a result of multiple divisions, reduced morphological and biochemical (creatine kinase, CK) differentiation were observed. Furthermore, MPD cells had significantly increased cells in the S and decreased cells in the G1 phases of the cell cycle versus CON, following serum withdrawal. These results suggest continued cycling rather than G1 exit and thus reduced differentiation (myotube atrophy) occurs in MPD muscle cells. These changes were underpinned by significant reductions in transcript expression of: IGF-I and myogenic regulatory factors (myoD and myogenin) together with elevated IGFBP5. Signaling studies showed that decreased differentiation in MPD was associated with decreased phosphorylation of Akt, and with later increased phosphorylation of JNK1/2. Chemical inhibition of JNK1/2 (SP600125) in MPD cells increased IGF-I expression (non-significantly), however, did not enhance differentiation. This study provides a potential model and molecular mechanisms for deterioration in differentiation capacity in skeletal muscle cells as a consequence of multiple population doublings that would potentially contribute to the ageing process. J. Cell. Biochem. 112: 3773–3785, 2011. © 2011 Wiley Periodicals, Inc.

Get access to the full text of this article

Ancillary