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Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture systemThe growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.
Document ID
20040112671
Document Type
Reprint (Version printed in journal)
Authors
Torgan, C. E. (Duke University Medical Center Durham, United States)
Burge, S. S.
Collinsworth, A. M.
Truskey, G. A.
Kraus, W. E.
Date Acquired
August 21, 2013
Publication Date
September 1, 2000
Publication Information
Publication: Medical & biological engineering & computing
Volume: 38
Issue: 5
ISSN: 0140-0118
Subject Category
Aerospace Medicine
Funding Number(s)
CONTRACT_GRANT: AR-08231
CONTRACT_GRANT: N01-HD-2-3144
Distribution Limits
Public
Copyright
Other
Keywords
Non-NASA Center
NASA Discipline Musculoskeletal