IGF-I and vitamin C promote myogenic differentiation of mouse and human skeletal muscle cells at low temperatures |
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Authors: | Shima Ai Pham Jennifer Blanco Erica Barton Elisabeth R Sweeney H Lee Matsuda Ryoichi |
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Affiliation: | aDepartment of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 309A Building 15, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan;bDepartment of Physiology, University of Pennsylvania School of Medicine, B701 Richards Building, 3700 Hamilton Walk, Philadelphia, PA 19104, USA;cDepartment of Anatomy and Cell Biology, University of Pennsylvania School of Dental Medicine, 441 Levy Building, 240 S. 40th Street, Philadelphia, PA 19104, USA |
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Abstract: | In a previous study investigating the effects of low temperature on skeletal muscle differentiation, we demonstrated that C2C12 mouse myoblasts cultured at 30 °C do not express myogenin, a myogenic regulatory factor (MRF), or fuse into multinucleated myotubes. At this low temperature, the myoblasts continuously express Id3, a negative regulator of MRFs, and do not upregulate muscle-specific microRNAs. In this study, we examined if insulin-like growth factor-I (IGF-I) and a stable form of vitamin C (L-ascorbic acid phosphate) could alleviate the low temperature-induced inhibition of myogenic differentiation in C2C12 cells. Although the addition of either IGF-I or vitamin C alone could promote myogenin expression in C2C12 cells at 30 °C, elongated multinucleated myotubes were not formed unless both IGF-I and vitamin C were continuously administered. In human skeletal muscle cells, low temperature-induced blockage of myogenic differentiation was also ameliorated by exogenous IGF-I and vitamin C. In addition, we demonstrated that satellite cells of IGF-I overexpressing transgenic mice in single-fiber culture expressed myogenin at a higher level than those of wild-type mice at 30 °C. This study suggests that body temperature plays an important role in myogenic differentiation of endotherms, but the sensitivity to low temperature could be buffered by certain factors in vivo, such as IGF-I and vitamin C. |
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Keywords: | Abbreviations: DM, differentiation medium DMEM, Dulbecco's modified Eagle's medium ECM, extracellular matrix EDL, extensor digitorum longus FBS, fetal bovine serum IGF, insulin-like growth factor MEM, minimal essential medium miRNA, microRNA MRF, myogenic regulatory factor MyHC, myosin heavy chain PBS, phosphate-buffered saline SEM, standard error of the mean VC, vitamin C WT, wild-type |
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