Dependence of in vitro myogenesis on a trophic protein present in chicken embryo extract.

Abstract

A trophic protein (sciatin) purified from sciatic nerves has been shown to enhance the morphological development and to promote the maintenance of skeletal muscle cells in vitro [Markelonis, G. J. & Oh, T. H. (1979) Proc. Natl. Acad. Sci. USA 76, 2470-2474]. We have elicited a specific antiserum against purified sciatin in rabbits. By using this antiserum, we have examined whether sciatin is also required for the initial differentiation of avian myogenic cells in vitro. Sciatin was found to be a component of chicken embryo extract, a constituent of culture medium required for myogenesis in vitro, by an immunodiffusion assay and by NaDodSO4 gel electrophoresis of immunoprecipitates. The removal of sciatin from chicken embryo extracts by immunoprecipitation with antiserum against sciatin completely inhibited myogenesis. When myogenic cells were grown in culture medium from which sciatin had been removed, the cells failed to differentiate beyond the myoblast stage. However, when sciatin (25 microgram/ml) was added to the sciatin-absorbed culture medium, normal myogenesis ensued. Furthermore, myogenic cells underwent normal myogenesis in the absence of embryo extract if sciatin (25 microgram/ml) was added to the culture medium. These results demonstrate that sciatin is the component of chicken embryo extract required for myogenesis and that the protein influences the initial differentiation of myogenic cells in vitro.