title: Isolation and validation of human prepubertal skeletal muscle cells: maturation and metabolic effects of IGF-I, IGFBP-3 and TNFalpha.
abstract: We have developed a primary skeletal muscle cell culture model derived from normal prepubertal children to investigate the effects of insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-3 (IGFBP-3) and tumour necrosis factor alpha (TNFalpha) on growth, differentiation and metabolism. Cells of myoblast lineage were characterized morphologically by desmin staining and differentiated successfully into multinucleated myotubes. Differentiation was confirmed biochemically by an increase in creatine kinase (CK) activity and IGFBP-3 secretion over time. IGF-I promoted whilst TNFalpha inhibited myoblast proliferation, differentiation and IGFBP-3 secretion. IGF-I partially rescued the cells from the inhibiting effects of TNFalpha. Compared to adult myoblast cultures, children's skeletal muscle cells demonstrated higher basal and day 7 CK activities, increased levels of IGFBP-3 secretion, diminished IGF-I/TNFalpha action and absence of the inhibitory effect of exogenous IGFBP-3 on differentiation. Additional studies demonstrated that TNFalpha increased basal glucose transport via GLUT1, nitric oxide synthase and p38MAPK-dependent mechanisms. These studies provide baseline data to study the interactivity effects of growth factors and cytokines on differentiation and metabolism in muscle in relation to important metabolic disorders such as obesity, type II diabetes or chronic wasting diseases.
authors.affiliation: Department of Exercise and Sport Science, Manchester Metropolitan University, Hassall Road, Alsager, UK. castewart@mmu.ac.uk.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Grohmann
authors.forename: Emily
authors.initials: E
authors.lastname: Foulstone
authors.forename: Gavin
authors.initials: G
authors.lastname: Welsh
authors.forename: Jeff
authors.initials: J
authors.lastname: Holly
authors.forename: Julian
authors.initials: J
authors.lastname: Shield
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Crowne
authors.forename: Claire
authors.initials: C
authors.lastname: Stewart
mesh_terms: D000328:Adult; D000367:Age Factors; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003402:Creatine Kinase / Q000378:metabolism; D003893:Desmin / Q000032:analysis; D005260:Female; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007223:Infant; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D008297:Male; D018485:Muscle Fibers, Skeletal / Q000187:drug effects* / Q000502:physiology; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000502:physiology; D032448:Myoblasts, Skeletal / Q000187:drug effects* / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D003893:Desmin; D018972:Insulin-Like Growth Factor Binding Protein 3; D014409:Tumor Necrosis Factor-alpha; D007334:Insulin-Like Growth Factor I; D019001:Nitric Oxide Synthase; D048051:p38 Mitogen-Activated Protein Kinases; D003402:Creatine Kinase; D005947:Glucose