title: Differentiation markers of mouse C2C12 and rat L6 myogenic cell lines and the effect of the differentiation medium.
abstract: The differentiation grade of cells in culture is dependent on the composition of the culture medium. Two commonly used myogenic cell lines, mouse C2C12 and rat L6, usually differentiate at a low concentration of horse serum. In this study we compared the effect of horse serum with a medium containing a low percentage of Ultroser G and rat brain extract. The maturation grade was evaluated on the basis of various biochemical, (immuno)histochemical and cell-physiological parameters. Substitution of horse serum by Ultroser G and rat brain extract during the differentiation phase resulted in a higher maturation grade of the myotubes of both cell lines, on the basis of creatine kinase activity and the diameter of the myotubes. In addition, the C2C12 myotubes display cross-striation, contain a higher percentage of creatine kinase muscle-specific isoenzyme MM, show a ninefold increase in acetylcholine receptor (AChR) clusters, form a continuous basement membrane, and have a lower resting cytosolic Ca2+ concentration. L6 myotubes show a fivefold increase in AChR clusters and a twofold increase in the expression of the mRNA of the epsilon-subunit of AChR.C2C12 cells show spontaneous contraction and response of cytosolic Ca2+ to various stimulants in contrast to L6 cells which do not. These studies established that the Ultroser G/brain extract medium leads to a higher differentiation grade of both cell lines, but parameters appropriate for use as differentiation markers appear to differ between both cell lines.
authors.affiliation: Department of Biochemistry, University of Nijmegen, The Netherlands.
authors.forename: G L
authors.initials: GL
authors.lastname: Portiér
authors.forename: A G
authors.initials: AG
authors.lastname: Benders
authors.forename: A
authors.initials: A
authors.lastname: Oosterhof
authors.forename: J H
authors.initials: JH
authors.lastname: Veerkamp
authors.forename: T H
authors.initials: TH
authors.lastname: van Kuppevelt
mesh_terms: D000818:Animals; D001798:Blood Proteins; D001802:Blood Substitutes; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002460:Cell Line; D003470:Culture Media; D051379:Mice; D018482:Muscle, Skeletal / Q000473:pathology*; D009930:Organic Chemicals; D051381:Rats
chemical_list: D001798:Blood Proteins; D001802:Blood Substitutes; D003470:Culture Media; D009930:Organic Chemicals; C044609:Ultroser G