Improved medium with EGF and BSA for differentiated human skeletal muscle cells

Epidermal growth factor (EGF) and bovine serum albumin (BSA) are both required for serum‐free clonal growth of human muscle satallite cells (HMSC). However, neither inhibits differentiation of HMSC, and when both are added to a minimal serum‐free differentiation medium, they enhance survival and maintenance of human myotubes. A combination of 10 ng/mL EGF and 0.5 mg/mL BSA, added to MCDB 120 plus 10 μg/mL insulin, increasesboth total protein per dish and total creatine kinase activity, and keeps the myotubes in good condition for a longer period of time. The myotubes become cross‐striated and exhibit frequent spontaneous twitching. Substantial amounts of neonatal myosin heavy chain and the MM isozyme of creatine kinase are expressed, together with detectable amounts of adult fast myosin heavy chain. With regular feeding, these cultures can be maintained for at least 3 weeks with no overgrowth by mononucleate cells, and with far less degeneration than with insulin as the only supplement.

[1]  R. G. Ham,et al.  Improved media for rapid clonal growth of normal human skeletal muscle satellite cells. , 1990, Advances in experimental medicine and biology.

[2]  L. Boxhorn,et al.  Regulation of skeletal muscle satellite cell proliferation and differentiation by transforming growth factor‐beta, insulin‐like growth factor I, and fibroblast growth factor , 1989, Journal of cellular physiology.

[3]  B. Olwin,et al.  Cell surface fibroblast growth factor and epidermal growth factor receptors are permanently lost during skeletal muscle terminal differentiation in culture , 1988, The Journal of cell biology.

[4]  H. Blau,et al.  Fast muscle fibers are preferentially affected in Duchenne muscular dystrophy , 1988, Cell.

[5]  J. Florini.  Hormonal control of muscle growth , 1987, Muscle & nerve.

[6]  C. Clegg,et al.  Growth factor control of skeletal muscle differentiation: commitment to terminal differentiation occurs in G1 phase and is repressed by fibroblast growth factor , 1987, The Journal of cell biology.

[7]  H. Blau,et al.  Developmental progression of myosin gene expression in cultured muscle cells , 1986, Cell.

[8]  L. Leinwand,et al.  Characterization of diverse forms of myosin heavy chain expressed in adult human skeletal muscle. , 1986, Nucleic acids research.

[9]  W. Engel,et al.  Fibroblast growth factor, epidermal growth factor and insulin exert a neuronal-like influence on acetylcholine receptors in aneurally cultured human muscle , 1985, Neuroscience Letters.

[10]  V. Askanas,et al.  Synergistic influence of polypeptide growth factors on cultured human muscle. , 1985, Archives of neurology.

[11]  C. Pinset,et al.  Induction of myogenic differentiation in serum-free medium does not require DNA synthesis. , 1985, Developmental biology.

[12]  D. Yaffe,et al.  A myogenic cell line with altered serum requirements for differentiation. , 1977, Differentiation; research in biological diversity.