Extracellular Matrix, Growth Factors, Genetics: Their Influence on Cell Proliferation and Myotube Formation in Primary Cultures of Adult Mouse Skeletal Muscle

Abstract

Cell proliferation and myotube formation in response to growth factors on various extracellular matrices (ECM) were investigated in primary skeletal muscle cultures from adult SJL/J and BALB/c mice. There was no difference between the rates of proliferation from primary cultures of SJL/J and Balb/c mice measured at 48 h in response to a range of concentrations of PDGF-AA, -AB, -BB, TGFβ1, or LIF (added at 24 h). SJL/J primary cultures were more responsive to bFGF (which was the most potent mitogen) than were BALB/c cultures. Comparison of dose response curves to bFGF and TGFβ1 grown on gelatin or Matrigel showed that the nature of the ECM did not have a significant affect. More myotubes formed at 4 days in SJL/J than in parallel BALB/c cultures on gelatin or Matrigel (P < 0.05). On gelatin more myotubes with 4 or more nuclei were formed in cultures from SJL/J than BALB/c muscle (P < 0.05); however, on Matrigel these myotubes occurred with similar frequency. Myotube formation examined in BALB/c muscle cultures grown on collagen IV, entactin-free laminin, and fibronectin showed that none of these ECM components alone supported large myotube formation (4 or more nuclei) as well as did Matrigel, although fibronectin was as effective as Matrigel with respect to the total number of myotubes formed. Parallel experiments carried out using the myogenic H-2Kb(27) cell line showed similar effects with the exception of laminin which enhanced large myotube formation and desmin expression in the H.2Kb(27) but not in the primary muscle cultures. The greater sensitivity in mitogenic response to bFGF and the more extensive myotube formation seen in SJL/J compared with BALB/c cultures in vitro reflects the superior capacity for muscle regeneration of SJL/J mice in vivo.