Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1982 Feb 1;92(2):462–470. doi: 10.1083/jcb.92.2.462

Changes in the components of extracellular matrix and in growth properties of cultured aortic smooth muscle cells upon ascorbate feeding

PMCID: PMC2112078  PMID: 7061590

Abstract

Culture conditions can modify the composition of the extracellular matrix of cultured calf aortas smooth muscle cells. In the absence of ascorbate the major components of the matrix are microfibrillar proteins; deposition of collagen occurs upon ascorbate supplementation and, with increased time of exposure of cells to ascorbate, collagen becomes the dominant protein of the extracellular matrix (greater than 80%). Collagen accumulation follows a sigmoidal time-course, suggesting that it is a cooperative phenomenon. Covalent crosslinks are not required for collagen accumulation in the matrix. Microfibrillar proteins and increased amounts of proteoglycans and fibronectin accumulate concurrently with collagen but elastin deposition was not observed either with or without ascorbate feeding. Addition of ascorbate leads to a general stimulation of incorporation of [14C]proline into cellular protein and to changes in cell growth parameters and morphology: cell-doubling time decreases from 62 to 47 h and plating efficiency increases approximately fourfold. We conclude that the composition of the extracellular matrix assembled by cultured cells is subject to experimental manipulation and that changes in endogenously deposited matrix may have significant effects on cellular functions.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Balian G., Click E. M., Bornstein P. Location of a collagen-binding domain in fibronectin. J Biol Chem. 1980 Apr 25;255(8):3234–3236. [PubMed] [Google Scholar]
  2. Bienkowski R. S., Engels C. J. Measurement of intracellular collagen degradation. Anal Biochem. 1981 Sep 15;116(2):414–424. doi: 10.1016/0003-2697(81)90382-1. [DOI] [PubMed] [Google Scholar]
  3. Blumenfeld O. O., Puglia K. V. Preparation of cyanogen bromide fragments of MM, NN, and MN glycoproteins (glycophorins) from human erythrocyte membranes of single donors. Biochim Biophys Acta. 1979 Jul 25;579(1):95–106. doi: 10.1016/0005-2795(79)90090-4. [DOI] [PubMed] [Google Scholar]
  4. Burke J. M., Ross R. Collagen synthesis by monkey arterial smooth muscle cells during proliferation and quiescence in culture. Exp Cell Res. 1977 Jul;107(2):387–395. doi: 10.1016/0014-4827(77)90360-3. [DOI] [PubMed] [Google Scholar]
  5. Burke J. M., Ross R. Synthesis of connective tissue macromolecules by smooth muscle. Int Rev Connect Tissue Res. 1979;8:119–157. doi: 10.1016/b978-0-12-363708-6.50010-2. [DOI] [PubMed] [Google Scholar]
  6. Cham B. E., Knowles B. R. A solvent system for delipidation of plasma or serum without protein precipitation. J Lipid Res. 1976 Mar;17(2):176–181. [PubMed] [Google Scholar]
  7. Cliff W. J. The ultrastructure of aortic elastica as revealed by prolonged treatment with OsO 4 . Exp Mol Pathol. 1971 Oct;15(2):220–229. doi: 10.1016/0014-4800(71)90101-8. [DOI] [PubMed] [Google Scholar]
  8. Fisher-Dzoga K., Jones R. M., Vesselinovitch D., Wissler R. W. Ultrastructural and immunohistochemical studies of primary cultures of aortic medial cells. Exp Mol Pathol. 1973 Apr;18(2):162–176. doi: 10.1016/0014-4800(73)90014-2. [DOI] [PubMed] [Google Scholar]
  9. Gelman R. A., Williams B. R., Piez K. A. Collagen fibril formation. Evidence for a multistep process. J Biol Chem. 1979 Jan 10;254(1):180–186. [PubMed] [Google Scholar]
  10. Gospodarowicz D., Ill C. R. Do plasma and serum have different abilities to promote cell growth? Proc Natl Acad Sci U S A. 1980 May;77(5):2726–2730. doi: 10.1073/pnas.77.5.2726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gospodarowicz D., Vlodavsky I., Savion N. The extracellular matrix and the control of proliferation of vascular endothelial and vascular smooth muscle cells. J Supramol Struct. 1980;13(3):339–372. doi: 10.1002/jss.400130307. [DOI] [PubMed] [Google Scholar]
  12. Hajek A. S., Solursh M. The effect of ascorbic acid on growth and synthesis of matrix components by cultured chick embryo chondrocytes. J Exp Zool. 1977 Jun;200(3):377–388. doi: 10.1002/jez.1402000308. [DOI] [PubMed] [Google Scholar]
  13. Hayman E. G., Ruoslahti E. Distribution of fetal bovine serum fibronectin and endogenous rat cell fibronectin in extracellular matrix. J Cell Biol. 1979 Oct;83(1):255–259. doi: 10.1083/jcb.83.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kahn P., Shin S. I. Cellular tumorigenicity in nude mice. Test of associations among loss of cell-surface fibronectin, anchorage independence, and tumor-forming ability. J Cell Biol. 1979 Jul;82(1):1–16. doi: 10.1083/jcb.82.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kleinman H. K., Klebe R. J., Martin G. R. Role of collagenous matrices in the adhesion and growth of cells. J Cell Biol. 1981 Mar;88(3):473–485. doi: 10.1083/jcb.88.3.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LANSING A. I., ROSENTHAL T. B., ALEX M., DEMPSEY E. W. The structure and chemical characterization of elastic fibers as revealed by elastase and by electron microscopy. Anat Rec. 1952 Dec;114(4):555–575. doi: 10.1002/ar.1091140404. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Lichtenstein J. R., Byers P. H., Smith B. D., Martin G. R. Identification of the collagenous proteins synthesized by cultured cells from human skin. Biochemistry. 1975 Apr 22;14(8):1589–1594. doi: 10.1021/bi00679a007. [DOI] [PubMed] [Google Scholar]
  20. Liotta L. A., Vembu D., Kleinman H. K., Martin G. R., Boone C. Collagen required for proliferation of cultured connective tissue cells but not their transformed counterparts. Nature. 1978 Apr 13;272(5654):622–624. doi: 10.1038/272622a0. [DOI] [PubMed] [Google Scholar]
  21. Perkins M. E., Ji T. H., Hynes R. O. Cross-linking of fibronectin to sulfated proteoglycans at the cell surface. Cell. 1979 Apr;16(4):941–952. doi: 10.1016/0092-8674(79)90109-0. [DOI] [PubMed] [Google Scholar]
  22. Rennard S. I., Wind M. L., Hewitt A. T., Kleinman H. K. Effect of collagen and cell shape on binding of fibronectin to cells. Arch Biochem Biophys. 1981 Jan;206(1):205–212. doi: 10.1016/0003-9861(81)90082-5. [DOI] [PubMed] [Google Scholar]
  23. Rojkind M., Gatmaitan Z., Mackensen S., Giambrone M. A., Ponce P., Reid L. M. Connective tissue biomatrix: its isolation and utilization for long-term cultures of normal rat hepatocytes. J Cell Biol. 1980 Oct;87(1):255–263. doi: 10.1083/jcb.87.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rojkind M., Giambrone M. A., Biempica L. Collagen types in normal and cirrhotic liver. Gastroenterology. 1979 Apr;76(4):710–719. [PubMed] [Google Scholar]
  25. Ross R. The elastic fiber. J Histochem Cytochem. 1973 Mar;21(3):199–208. doi: 10.1177/21.3.199. [DOI] [PubMed] [Google Scholar]
  26. Schwartz E., Adamany A. M., Blumenfeld O. O. Extracellular proteins of the calf aortic media smooth muscle cells in culture. Biochim Biophys Acta. 1980 Aug 21;624(2):531–544. doi: 10.1016/0005-2795(80)90094-x. [DOI] [PubMed] [Google Scholar]
  27. Scott-Burden T., Davies P. J., Gevers W. Elastin biosynthesis by smooth muscle cells cultured under scorbutic conditions. Biochem Biophys Res Commun. 1979 Dec 14;91(3):739–746. doi: 10.1016/0006-291x(79)91942-9. [DOI] [PubMed] [Google Scholar]
  28. Siegel R. C., Pinnell S. R., Martin G. R. Cross-linking of collagen and elastin. Properties of lysyl oxidase. Biochemistry. 1970 Nov 10;9(23):4486–4492. doi: 10.1021/bi00825a004. [DOI] [PubMed] [Google Scholar]
  29. Soltoff-Schiller B., Goldfischer S., Adamany A. M., Wolinsky H. Endocytosis by vascular smooth muscle cells in vivo and in vitro. Roles or vesicles and lysosomes. Am J Pathol. 1976 Apr;83(1):45–60. [PMC free article] [PubMed] [Google Scholar]
  30. VAN DEN HOOFF A., LEVENE C. I., GROSS J. Morphologic evidence for collagen changes in chick embryos treated with beta-aminopropionitrile. J Exp Med. 1959 Dec 1;110:1017–1022. doi: 10.1084/jem.110.6.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. de Clerck Y. A., Jones P. A. The effect of ascorbic acid on the nature and production of collagen and elastin by rat smooth-muscle cells. Biochem J. 1980 Jan 15;186(1):217–225. doi: 10.1042/bj1860217. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES