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. 1984 Jun;81(11):3429–3432. doi: 10.1073/pnas.81.11.3429

Growth hormone promotes the differentiation of myoblasts and preadipocytes generated by azacytidine treatment of 10T1/2 cells.

B T Nixon, H Green
PMCID: PMC345521  PMID: 6203122

Abstract

Previous studies have shown that growth hormone promotes the differentiation of preadipose 3T3 cells into adipose cells. This action of the hormone is not mediated by insulin-like growth factor 1 (IGF-1), but is exerted directly on the preadipocytes. In order to determine whether growth hormone promotes the differentiation of other mesenchymal cell types, we have examined its effect on the formation of multinucleated muscle cells from myoblasts. Clone mu cells, generated from line 10T1/2 by treatment with azacytidine, were known to be capable of undergoing myogenesis. When these cells were cultivated in a medium not able alone to support their differentiation, the addition of physiological concentrations of growth hormone strongly promoted myogenesis. When fetal calf serum was used instead of growth hormone to promote myogenesis, the effectiveness of the serum was reduced by the addition of a specific antiserum to growth hormone. IGF-1 could not substitute for growth hormone in promoting myogenesis. When another 10T1/2 line not ordinarily able to form myotubes was subjected to treatment with 5-azacytidine, it was subsequently able to give rise to muscle and adipose cell colonies; the number of such colonies was much increased by the presence of growth hormone. We have not been able to demonstrate a similar dependence on growth hormone for myogenesis by the established lines L6E9 and C2.

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Selected References

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  1. Beach R. K., Kostyo J. L. Effect of growth hormone on the DNA content of muscles of young hypophysectomized rats. Endocrinology. 1968 Apr;82(4):882–884. doi: 10.1210/endo-82-4-882. [DOI] [PubMed] [Google Scholar]
  2. Cohen A., Buckingham M., Gros F. A modified assay procedure for revealing the M form of creatine kinase in cultured muscle cells. Exp Cell Res. 1978 Aug;115(1):201–206. doi: 10.1016/0014-4827(78)90417-2. [DOI] [PubMed] [Google Scholar]
  3. Eden S., Isaksson O. G., Madsen K., Friberg U. Specific binding of growth hormone to isolated chondrocytes from rabbit ear and epiphyseal plate. Endocrinology. 1983 Mar;112(3):1127–1129. doi: 10.1210/endo-112-3-1127. [DOI] [PubMed] [Google Scholar]
  4. Ewton D. Z., Florini J. R. Effects of the somatomedins and insulin on myoblast differentiation in vitro. Dev Biol. 1981 Aug;86(1):31–39. doi: 10.1016/0012-1606(81)90312-2. [DOI] [PubMed] [Google Scholar]
  5. Fagin K. D., Lackey S. L., Reagan C. R., DiGirolamo M. Specific binding of growth hormone by rat adipocytes. Endocrinology. 1980 Aug;107(2):608–615. doi: 10.1210/endo-107-2-608. [DOI] [PubMed] [Google Scholar]
  6. Gavin J. R., 3rd, Saltman R. J., Tollefsen S. E. Growth hormone receptors in isolated rat adipocytes. Endocrinology. 1982 Feb;110(2):637–643. doi: 10.1210/endo-110-2-637. [DOI] [PubMed] [Google Scholar]
  7. Gluckman P. D., Grumbach M. M., Kaplan S. L. The neuroendocrine regulation and function of growth hormone and prolactin in the mammalian fetus. Endocr Rev. 1981 Fall;2(4):363–395. doi: 10.1210/edrv-2-4-363. [DOI] [PubMed] [Google Scholar]
  8. Goeddel D. V., Heyneker H. L., Hozumi T., Arentzen R., Itakura K., Yansura D. G., Ross M. J., Miozzari G., Crea R., Seeburg P. H. Direct expression in Escherichia coli of a DNA sequence coding for human growth hormone. Nature. 1979 Oct 18;281(5732):544–548. doi: 10.1038/281544a0. [DOI] [PubMed] [Google Scholar]
  9. Goldspink D. F., Goldberg A. L. Influence of pituitary growth hormone on DNA synthesis in rat tissues. Am J Physiol. 1975 Jan;228(1):302–309. doi: 10.1152/ajplegacy.1975.228.1.302. [DOI] [PubMed] [Google Scholar]
  10. Herington A. C., Phillips L. S., Daughaday W. H. Pituitary regulation of human growth hormone binding sites in rat liver membranes. Metabolism. 1976 Mar;25(3):341–353. doi: 10.1016/0026-0495(76)90092-5. [DOI] [PubMed] [Google Scholar]
  11. Holder A. T., Spencer E. M., Preece M. A. Effect of bovine growth hormone and a partially pure preparation of somatomedin on various growth parameters in hypopituitary dwarf mice. J Endocrinol. 1981 May;89(2):275–282. doi: 10.1677/joe.0.0890275. [DOI] [PubMed] [Google Scholar]
  12. Isaksson O. G., Jansson J. O., Gause I. A. Growth hormone stimulates longitudinal bone growth directly. Science. 1982 Jun 11;216(4551):1237–1239. doi: 10.1126/science.7079756. [DOI] [PubMed] [Google Scholar]
  13. Isaksson O. G., Schwartz J., Kostyo J. L., Reagan C. R. Use of antibodies to modify the effect of growth hormone on sugar transport in rat diaphragm muscle. Endocrinology. 1979 Aug;105(2):452–458. doi: 10.1210/endo-105-2-452. [DOI] [PubMed] [Google Scholar]
  14. Jones P. A., Taylor S. M. Cellular differentiation, cytidine analogs and DNA methylation. Cell. 1980 May;20(1):85–93. doi: 10.1016/0092-8674(80)90237-8. [DOI] [PubMed] [Google Scholar]
  15. Klapper D. G., Svoboda M. E., Van Wyk J. J. Sequence analysis of somatomedin-C: confirmation of identity with insulin-like growth factor I. Endocrinology. 1983 Jun;112(6):2215–2217. doi: 10.1210/endo-112-6-2215. [DOI] [PubMed] [Google Scholar]
  16. Kostyo J. L., Isaksson O. Growth hormone and the regulation of somatic growth. Int Rev Physiol. 1977;13:255–274. [PubMed] [Google Scholar]
  17. Kuri-Harcuch W., Green H. Adipose conversion of 3T3 cells depends on a serum factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6107–6109. doi: 10.1073/pnas.75.12.6107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lesniak M. A., Gorden P., Roth J., Gavin J. R., 3rd Binding of 125I-human growth hormone to specific receptors in human cultured lymphocytes. Characterization of the interaction and a sensitive radioreceptor assay. J Biol Chem. 1974 Mar 25;249(6):1661–1667. [PubMed] [Google Scholar]
  19. Madsen K., Friberg U., Roos P., Edén S., Isaksson O. Growth hormone stimulates the proliferation of cultured chondrocytes from rabbit ear and rat rib growth cartilage. Nature. 1983 Aug 11;304(5926):545–547. doi: 10.1038/304545a0. [DOI] [PubMed] [Google Scholar]
  20. Martial J. A., Hallewell R. A., Baxter J. D., Goodman H. M. Human growth hormone: complementary DNA cloning and expression in bacteria. Science. 1979 Aug 10;205(4406):602–607. doi: 10.1126/science.377496. [DOI] [PubMed] [Google Scholar]
  21. Morikawa M., Green H., Lewis U. J. Activity of human growth hormone and related polypeptides on the adipose conversion of 3T3 cells. Mol Cell Biol. 1984 Feb;4(2):228–231. doi: 10.1128/mcb.4.2.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morikawa M., Nixon T., Green H. Growth hormone and the adipose conversion of 3T3 cells. Cell. 1982 Jul;29(3):783–789. doi: 10.1016/0092-8674(82)90440-8. [DOI] [PubMed] [Google Scholar]
  23. Nadal-Ginard B. Commitment, fusion and biochemical differentiation of a myogenic cell line in the absence of DNA synthesis. Cell. 1978 Nov;15(3):855–864. doi: 10.1016/0092-8674(78)90270-2. [DOI] [PubMed] [Google Scholar]
  24. Nixon T., Green H. Contribution of growth hormone to the adipogenic activity of serum. Endocrinology. 1984 Feb;114(2):527–532. doi: 10.1210/endo-114-2-527. [DOI] [PubMed] [Google Scholar]
  25. Nixon T., Green H. Properties of growth hormone receptors in relation to the adipose conversion of 3T3 cells. J Cell Physiol. 1983 Jun;115(3):291–296. doi: 10.1002/jcp.1041150312. [DOI] [PubMed] [Google Scholar]
  26. Olson K. C., Fenno J., Lin N., Harkins R. N., Snider C., Kohr W. H., Ross M. J., Fodge D., Prender G., Stebbing N. Purified human growth hormone from E. coli is biologically active. Nature. 1981 Oct 1;293(5831):408–411. doi: 10.1038/293408a0. [DOI] [PubMed] [Google Scholar]
  27. Rinderknecht E., Humbel R. E. Polypeptides with nonsuppressible insulin-like and cell-growth promoting activities in human serum: isolation, chemical characterization, and some biological properties of forms I and II. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2365–2369. doi: 10.1073/pnas.73.7.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Savage C. R., Jr, Cohen S. Epidermal growth factor and a new derivative. Rapid isolation procedures and biological and chemical characterization. J Biol Chem. 1972 Dec 10;247(23):7609–7611. [PubMed] [Google Scholar]
  29. Schoenle E., Zapf J., Humbel R. E., Froesch E. R. Insulin-like growth factor I stimulates growth in hypophysectomized rats. Nature. 1982 Mar 18;296(5854):252–253. doi: 10.1038/296252a0. [DOI] [PubMed] [Google Scholar]
  30. Sedmak J. J., Grossberg S. E. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem. 1977 May 1;79(1-2):544–552. doi: 10.1016/0003-2697(77)90428-6. [DOI] [PubMed] [Google Scholar]
  31. Seeburg P. H., Shine J., Martial J. A., Ivarie R. D., Morris J. A., Ullrich A., Baxter J. D., Goodman H. M. Synthesis of growth hormone by bacteria. Nature. 1978 Dec 21;276(5690):795–798. doi: 10.1038/276795a0. [DOI] [PubMed] [Google Scholar]
  32. Stewart J. K., Reagan C. R., Kostyo J. L. Correlation between the biological and immunological activities of growth hormone circulating in rats bearing MtTW15 tumors. Endocrinology. 1977 May;100(5):1376–1383. doi: 10.1210/endo-100-5-1376. [DOI] [PubMed] [Google Scholar]
  33. Taylor S. M., Jones P. A. Changes in phenotypic expression in embryonic and adult cells treated with 5-azacytidine. J Cell Physiol. 1982 May;111(2):187–194. doi: 10.1002/jcp.1041110210. [DOI] [PubMed] [Google Scholar]
  34. Taylor S. M., Jones P. A. Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5-azacytidine. Cell. 1979 Aug;17(4):771–779. doi: 10.1016/0092-8674(79)90317-9. [DOI] [PubMed] [Google Scholar]
  35. Tsushima T., Friesen H. G. Radioreceptor assay for growth hormone. J Clin Endocrinol Metab. 1973 Aug;37(2):334–337. doi: 10.1210/jcem-37-2-334. [DOI] [PubMed] [Google Scholar]
  36. Van den Brande J. L., van Buul-Offers S. Effect of growth hormone and peptide fractions containing somatomedin activity on growth and cartilage metabolism of Snell dwarfmice. Acta Endocrinol (Copenh) 1979 Oct;92(2):242–257. doi: 10.1530/acta.0.0920242. [DOI] [PubMed] [Google Scholar]
  37. Yaffe D. Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proc Natl Acad Sci U S A. 1968 Oct;61(2):477–483. doi: 10.1073/pnas.61.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yaffe D., Saxel O. Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature. 1977 Dec 22;270(5639):725–727. doi: 10.1038/270725a0. [DOI] [PubMed] [Google Scholar]
  39. Zapf J., Froesch E. R., Humbel R. E. The insulin-like growth factors (IGF) of human serum: chemical and biological characterization and aspects of their possible physiological role. Curr Top Cell Regul. 1981;19:257–309. doi: 10.1016/b978-0-12-152819-5.50024-5. [DOI] [PubMed] [Google Scholar]

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