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In vitro characterization of proliferation and differentiation of trout satellite cells.
MedLine Citation:
PMID:  21086139     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
Fish satellite cells have been extracted from various species, but the myogenic characteristics of these cells in culture remain largely unknown. We show here that 60%-70% of the adherent cells are myogenic based on their immunoreactivity for the myogenic regulatory factor MyoD. In DMEM containing 10% fetal calf serum (FCS), trout myoblasts display rapid expression of myogenin (18% of myogenin-positive cells at day 2) combined with rapid fusion into myotubes (50% of myogenin-positive nuclei and 30% nuclei in myosin heavy chain [MyHC]-positive cells at day 7). These kinetics of differentiation are reminiscent of the behavior of fetal myoblasts in mammals. However, not all the myogenic cells differentiate; this subpopulation of cells might correspond to the previously named "reserve" cells. More than 90% of the BrdU-positive cells are also positive for MyoD, indicating that myogenic cells proliferate in vitro. By contrast, less than 1% of myogenin-positive cells are positive for BrdU suggesting that myogenin expression occurs only in post-mitotic cells. In order to maximize either the proliferation or the differentiation of cells, we have defined new culture conditions based on the use of a proliferation medium (F10+10%FCS) and a differentiation medium (DMEM+2%FCS). Three days after switching the medium, the differentiation index (% MyHC-positive nuclei) is 40-fold higher than that in proliferation medium, whereas the proliferation index (% BrdU-positive nuclei) is three-fold lower. Stimulation of cell proliferation by insulin-like growth factor 1 (IGF1), IGF2, and FGF2 is greater in F10 medium. The characterization of these extracted muscle cells thus validates the use of this in vitro system of myogenesis in further studies of the myogenic activity of growth factors in trout.
Authors:
Jean Charles Gabillard; Nathalie Sabin; Gilles Paboeuf
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Publication Detail:
Type:  Journal Article     Date:  2010-11-18
Journal Detail:
Title:  Cell and tissue research     Volume:  342     ISSN:  1432-0878     ISO Abbreviation:  Cell Tissue Res.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-12-03     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0417625     Medline TA:  Cell Tissue Res     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  471-7     Citation Subset:  IM    
Affiliation:
INRA, UR1037 Station Commune de Recherches en Ichtyophysiologie Biodiversité et Environnement SCRIBE, Equipe Croissance et Qualité de la Chair de Poisson, Campus Beaulieu, 35000 Rennes, France. Jean-Charles.Gabillard@rennes.inra.fr
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