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. 1982 May 1;93(2):395–401. doi: 10.1083/jcb.93.2.395

Inhibition of myoblast differentiation in vitro by a protein isolated from liver cell medium

PMCID: PMC2112867  PMID: 7047537

Abstract

We have recently discovered that cells of Coon's Buffalo rat liver (BRL) line secrete a protein which is a potent inhibitor of skeletal myoblast differentiation in vitro. Using ion exchange and molecular exclusion chromatography, we have prepared this protein, which we designate "differentiation inhibitor" (DI), from the materials secreted by BRL cells maintained in serum-free medium. It is a relatively heat- stable protein which is inactivated by treatment with trypsin and mercaptoethanol and has an apparent molecular weight in the range 30,000--36,000. It exhibits no detectable mitogenic or lectin activity and differs from previously reported inhibitors of myoblast differentiation in several respects. It is active in all skeletal myoblast systems tested (Yaffe's L6 line as well as primary cultures of rat, chick, and Japanese quail myoblasts), and it blocks fusion, elevation of creatine kinase, and increased binding of alpha- bungarotoxin. Parallel fractionation of fetal bovine serum (FBS) and chick embryo extract (CEE) yields a peak of activity which similarly inhibits myoblast differentiation. We suggest that the differentiation inhibitor from BRL cells may correspond to the differentiation- inhibiting component(s) of FBS and CEE, and we call attention to the possibility that such a substance could play a role in embryonic growth of myoblasts and in satellite cell formation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Den H., Malinzak D. A. Isolation and properties of beta-D-galactoside-specific lectin from chick embryo thigh muscle. J Biol Chem. 1977 Aug 10;252(15):5444–5448. [PubMed] [Google Scholar]
  2. Erwin B. G., Stoscheck C. M., Florini J. R. A rapid fluorometric method for the estimation of DNA in cultured cells. Anal Biochem. 1981 Jan 15;110(2):291–294. doi: 10.1016/0003-2697(81)90194-9. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Florini J. R., Nicholson M. L., Dulak N. C. Effects of peptide anabolic hormones on growth of myoblasts in culture. Endocrinology. 1977 Jul;101(1):32–41. doi: 10.1210/endo-101-1-32. [DOI] [PubMed] [Google Scholar]
  5. Gartner T. K., Podleski T. R. Evidence that the types and specific activity of lectins control fusion of L6 myoblasts. Biochem Biophys Res Commun. 1976 Jun 21;70(4):1142–1149. doi: 10.1016/0006-291x(76)91022-6. [DOI] [PubMed] [Google Scholar]
  6. Knauer D. J., Smith G. L. Inhibition of biological activity of multiplication-stimulating activity by binding to its carrier protein. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7252–7256. doi: 10.1073/pnas.77.12.7252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Konigsberg I. R. Diffusion-mediated control of myoblast fusion. Dev Biol. 1971 Sep;26(1):133–152. doi: 10.1016/0012-1606(71)90113-8. [DOI] [PubMed] [Google Scholar]
  8. Konigsberg I. R. Skeletal myoblasts in culture. Methods Enzymol. 1979;58:511–527. doi: 10.1016/s0076-6879(79)58166-x. [DOI] [PubMed] [Google Scholar]
  9. Kuri-Harcuch W., Green H. Suppression of the adipose conversion of 3T3 cells by acidified serum. J Cell Physiol. 1981 Sep;108(3):455–460. doi: 10.1002/jcp.1041080320. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Linkhart T. A., Clegg C. H., Hauschika S. D. Myogenic differentiation in permanent clonal mouse myoblast cell lines: regulation by macromolecular growth factors in the culture medium. Dev Biol. 1981 Aug;86(1):19–30. doi: 10.1016/0012-1606(81)90311-0. [DOI] [PubMed] [Google Scholar]
  12. MAURO A. Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol. 1961 Feb;9:493–495. doi: 10.1083/jcb.9.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. MacBride R. G., Przybylski R. J. Purified lectin from skeletal muscle inhibits myotube formation in vitro. J Cell Biol. 1980 Jun;85(3):617–625. doi: 10.1083/jcb.85.3.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Merrill G. F., Florini J. R., Dulak N. C. Effects of multiplication stimulating activity (MSA) on AIB transport into myoblast and myotube cultures. J Cell Physiol. 1977 Nov;93(2):173–182. doi: 10.1002/jcp.1040930202. [DOI] [PubMed] [Google Scholar]
  15. Moses A. C., Nissley S. P., Passamani J., White R. M. Further characterization of growth hormone-dependent somatomedin-binding proteins in rat serum and demonstration of somatomedin-binding proteins produced by rat liver cells in culture. Endocrinology. 1979 Feb;104(2):536–546. doi: 10.1210/endo-104-2-536. [DOI] [PubMed] [Google Scholar]
  16. Moses A. C., Nissley S. P., Short P. A., Rechler M. M., White R. M., Knight A. B., Higa O. Z. Increased levels of multiplication-stimulating activity, an insulin-like growth factor, in fetal rat serum. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3649–3653. doi: 10.1073/pnas.77.6.3649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nowak T. P., Kobiler D., Roel L. E., Barondes S. H. Developmentally regulated lectin from embryonic chick pectoral muscle. Purification by affinity chromatography. J Biol Chem. 1977 Sep 10;252(17):6026–6030. [PubMed] [Google Scholar]
  18. Pierson R. W., Jr, Temin H. M. The partial purification from calf serum of a fraction with multiplication-stimulating activity for chicken fibroblasts in cell culture and with non-suppressible insulin-like activity. J Cell Physiol. 1972 Jun;79(3):319–330. doi: 10.1002/jcp.1040790302. [DOI] [PubMed] [Google Scholar]
  19. Podleski T. R., Greenberg I., Schlessinger J., Yamada K. M. Fibronectin delays the fusion of L6 myoblasts. Exp Cell Res. 1979 Sep;122(2):317–326. doi: 10.1016/0014-4827(79)90308-2. [DOI] [PubMed] [Google Scholar]
  20. Simpson D. L., Thorne D. R., Loh H. H. Lectins: endogenous carbohydrate-binding proteins from vertebrate tissues: functional role in recognition processes? Life Sci. 1978 Mar;22(9):727–748. doi: 10.1016/0024-3205(78)90242-4. [DOI] [PubMed] [Google Scholar]
  21. Slater C. R. Control of myogenesis in vitro by chick embryo extract. Dev Biol. 1976 Jun;50(2):264–284. doi: 10.1016/0012-1606(76)90151-2. [DOI] [PubMed] [Google Scholar]
  22. 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]

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