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Articles in PresS, published online ahead of print June 26, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00168.2002
Submitted on April 12, 2002
Accepted on May 23, 2002
1 Oral Growth and Developmental Biology, Hiroshima University, Hiroshima, Japan
* To whom correspondence should be addressed. E-mail: norihiko{at}hiroshima-u.ac.jp.
Myogenic differentiation in vitro has been usually viewed as being negatively controlled by serum mitogens. A depletion of critical serum componentsfrom medium has been considered to be essential for permanent withdrawal from the cell cycle and terminal differentiation of myoblasts. Removal of serum mitogens induces the expression of insulin-like growth factors (IGF)s, whereas it inhibits that of basic fibroblast growth factor (bFGF) and transforming growth factor (TGF)-ß in myoblasts. These responses of growth factors to medium conditioning seem to be well matched to their functions in proliferation/differentiation. In the present study, we showed that C2C12 myoblasts differentiated actively even in mitogen-rich medium and that this medium offered an advantage over mitogen-poor medium in terms of increasing differentiation. Our attention focused on endogenous growth factors as described above, especially IGFs in mitogen-rich medium. During differentiation, IGF-I and IGF-II mRNA levels increased, but bFGF and TGF-ß1 mRNA decreased. Differentiation was commensurable with IGF mRNA levels and suppressed by antisense oligodeoxynucleotides and neutralizing monoclonal antibodies against IGFs. These results suggest that an autocrine/paracrine loop of IGFs, bFGF and TGF-ß1 is active in proliferating and differentiating C2C12 cells without a depletion of serum and that endogenous IGFs actively override the negative control of differentiation by serum mitogens.
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