title: The biochemical and structural maturation of human skeletal muscle cells in culture: the effect of the serum substitute Ultroser G.
abstract: On the basis of the percentage creatine kinase-MM, human skeletal muscle cells cultured on growth and differentiation media containing the serum substitute Ultroser G reach a significantly higher maturation grade after 7 days of differentiation than cells cultured on serum-containing media. They also remain viable for longer periods. The myotubes are much longer, their nuclei are often localized in rows on the periphery, and they show cross-striation more frequently. The activities of creatine kinase, citrate synthase, cytochrome c oxidase, AMP deaminase, and phosphorylase are significantly higher. Extending the differentiation period to 3 weeks increases the maturation grade of the cultures and the activities of all the enzymes mentioned before, except phosphorylase. A correlation exists between the enzyme activities and the maturation grade of the muscle cells. The content of fatty acid-binding protein also increases significantly with the maturation grade in contrast to the palmitate oxidation rate. The AMP deaminase and creatine kinase activity and the percentage MM-type remain lower in cultured cells than in adult muscle and the hexokinase activity remains higher, but the other enzyme activities become comparable after 20 days of differentiation. The myotubes, derived from Ultroser G-containing culture media, show spontaneous contractions after 12 days and cross-striation after 20 days when immunostained for the M-subunit of creatine kinase. These cells possess clusters of acetylcholine receptors, but aggregation of desmin at the site of the clusters was never detectable. The possibility of cultivating muscle cells with a predictable maturation grade allows the study of muscle development and muscular diseases caused by differentiation defects or by deficiency of a maturation-dependent (iso)enzyme.
authors.affiliation: Department of Biochemistry, University of Nijmegen, The Netherlands.
authors.forename: A A
authors.initials: AA
authors.lastname: Benders
authors.forename: T H
authors.initials: TH
authors.lastname: van Kuppevelt
authors.forename: A
authors.initials: A
authors.lastname: Oosterhof
authors.forename: J H
authors.initials: JH
authors.lastname: Veerkamp
mesh_terms: D000659:AMP Deaminase / Q000378:metabolism; D000818:Animals; D001802:Blood Substitutes / Q000494:pharmacology*; D002352:Carrier Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002950:Citrate (si)-Synthase / Q000378:metabolism; D003402:Creatine Kinase / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology*; D003576:Electron Transport Complex IV / Q000378:metabolism; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D006801:Humans; D007150:Immunohistochemistry; D009132:Muscles / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009363:Neoplasm Proteins; D009930:Organic Chemicals; D025521:Tumor Suppressor Proteins
chemical_list: D001802:Blood Substitutes; D002352:Carrier Proteins; D003470:Culture Media; C107608:FABP7 protein, human; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D009930:Organic Chemicals; D025521:Tumor Suppressor Proteins; C044609:Ultroser G; D003576:Electron Transport Complex IV; D002950:Citrate (si)-Synthase; D003402:Creatine Kinase; D000659:AMP Deaminase