C2 and C2C12 murine skeletal myoblast models of atrophic and hypertrophic potential: Relevance to disease and ageing?

Authors

  • Adam P. Sharples,

    Corresponding author
    1. Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health, Manchester Metropolitan University, Manchester, UK
    • Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health, Manchester Metropolitan University, John Dalton Building, Oxford Road, Manchester M1 5GD, UK.
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  • Nasser Al-Shanti,

    1. Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health, Manchester Metropolitan University, Manchester, UK
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  • Claire E. Stewart

    1. Faculty of Science and Engineering, Institute for Biomedical Research into Human Movement and Health, Manchester Metropolitan University, Manchester, UK
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  • The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work.

Abstract

Reduced muscle mass and increased susceptibility to TNF-induced degradation accompany inflamed ageing and chronic diseases. Furthermore, C2 myoblasts display diminished differentiation and increased susceptibility to TNF-α-induced cell death versus subcloned C2C12 cells, providing relevant models to assess: differentiation (creatine kinase), growth (protein), death (trypan-blue) and anabolic/catabolic parameters (RT-PCR) over 72 h ± TNF-α (20 ng ml−1). At 48 and 72 h, respectively, larger myotubes and significantly higher CK activity (320.26 ± 6.82 vs. 30.71 ± 2.5, P < 0.05; 544.94 ± 27.7 vs. 39.4 ± 3.37 mU mg ml−1, P < 0.05), fold increases in myoD (21.45 ± 3.12 vs. 3.97 ± 1.76, P < 0.05; 31.07 ± 3.1 vs. 6.82 ± 1.93, P < 0.05) and myogenin mRNA (241.8 ± 40 vs. 36.80 ± 19.3, P < 0.05; 440 ± 100.5 vs. 201.1 ± 86, P < 0.05) were detected in C2C12 versus C2. C2C12 showed significant increases in IGF-I mRNA (243.05 ± 3.87 vs. 105.75 ± 21.95, P < 0.05), reduced proliferation and significantly lower protein expression (1.21 ± 0.28 vs. 1.79 ± 0.29 mg ml−1, P < 0.05) at 72 h versus C2 cells. Significant temporal reductions in C2C12 IGFBP2 mRNA (28.02 ± 15.44, 13.82 ± 8.07, 6.92 ± 4.37, P < 0.05) contrasted increases in C2s (4.31 ± 3.31, 13.02 ± 9.92, 82.9 ± 58.9, P < 0.05) at 0, 48 and 72 h, respectively. TNF-α increased cell death in C2s (2.67 ± 1.54%, 34.42 ± 5.39%, 29.71 ± 5.79% (0, 48, 72 h), P < 0.05), yet was without effect in C2C12s at 48 h but caused a small significant increase at 72 h (9.88 ± 4.02% (TNF-α) vs. 6.17 ± 0.749% (DM), 72 h). TNF-α and TNFRI mRNA were unchanged; however, larger reductions in IGF-I (8.2- and 7.5-fold vs. 4.5- and 4.1-fold (48, 72 h)), IGF-IR (2-fold vs. no-significant reduction (72 h)) and IGFBP5 (3.24 vs. 1.38 (48 h) and 2.21 vs. 1.71 (72 h), P < 0.05) mRNA were observed in C2 versus C2C12 with TNF-α. This investigation provides insight into regulators of altered basal hypertrophy and TNF-induced atrophy, providing a model for future investigation into therapeutic initiatives for ageing/wasting disorders. J. Cell. Physiol. 225: 240–250, 2010. © 2010 Wiley-Liss, Inc.

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