This report describes a phenotyping study of differentiating human skeletal muscle cells in tissue culture. Satellite cells (adult myoblasts), isolated from biopsy material, showed a proliferative behaviour in high-nutrition medium, but fused to form myotubes when grown in low-nutrition medium. The expression and structural organization of the intermediate filament proteins desmin and vimentin as well as the sarcomeric constituents alpha-actin, alpha-actinin, nebulin, myosin and especially titin during myofibrillogenesis in vitro, were studied by means of indirect immunofluorescence assays. The proliferating myoblasts contained both desmin and vimentin, alpha-actinin and the filamentous form of actin. Shortly after the change of medium, expression of titin, sarcomeric myosin and skeletal muscle alpha-actin was found in mononuclear cells in a diffuse, filamentous (titin, myosin, alpha-actin) or punctate (titin, myosin) pattern. Four to 10 days after the medium change, mature myotubes showed desmin, titin, alpha-actinin, nebulin, sarcomeric myosin and actin cross-striations, while vimentin was no longer detected. We conclude that human skeletal muscle cell cultures are an appropriate model system to study the molecular basis of myofibrillogenesis. Especially the presence of desmin in a striated fashion points to a high degree of maturation of the muscle cell cultures.

Stimulated by the rather sparse information in the literature on cellular changes induced by EHDP, we carried out electron microscopic investigations on young bone tissue and on de novo bone formation. Cellular changes could be observed during continuous administration of EHDP. The osteoblasts demonstrated temporary storing of crystalloid structures in the mitochondria, and atypical osteocytes showed persistent changes indicative of hyperactivity. The osteoclasts exhibited varying ultrastructural features with respect to the number and appearance of nuclei, Golgi, RER, and lysosomes. These changes under the influence of EHDP could be an indication of altered activity of the osteoclast. The possible interference of EHDP with bone cell metabolism is discussed.

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