Macrophage-released Factor Stimulates Selectively Myogenic Cells in Primary Muscle Culture

Myofibers are reconstituted by the proliferation and fusion of muscle precursor cells when skeletal muscle is injured. One of the critical events is the peak accumulation of macrophages after 48 hours at the damage site before the satellite cell proliferation. In addition to their well-known role as a scavenger cell, there is now direct evidence of a mitogenic role of macrophages in regenerating muscle. We have utilized an in vitro model to directly investigate and prove that macrophages increase myoblast growth not only of satellite cells, but also of primary myoblasts. Rat muscle cells were cultured in the presence or absence of exudate macrophages obtained by peritoneal washing after thioglycollate broth injection. Macrophage coculture increases several times the myoblasts/myotubes yield. This effect is particularly evident in muscle culture conditions in which fibroblast growth is predominant over myoblast proliferation, suggesting a myoblast selective mitogenic effect of macrophages. The results are confirmed by quantitative analyses of both DNA and skeletal muscle-specific-contractile proteins by gel electrophoresis and immunocytochemistry. Experiments with macrophage-conditioned media show this effect is mediated by soluble factors. This growth factor-like activity, which has been shown to be acid-stable and heat-labile, labile, exerts its effects not only on specialized satellite cells during muscle regeneration, but also has a broader mitotic activity on all myogenic cells. In view of the role of muscle regeneration in muscle diseases and of the perspectives offered by gene therapy via myoblasts, we strongly believe that our results open new opportunities in removing many of the clinical constraints associated with repair and cell transplantation.