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. 1979 Nov 1;83(2):468–486. doi: 10.1083/jcb.83.2.468

Vascular endothelial cells maintained in the absence of fibroblast growth factor undergo structural and functional alterations that are incompatible with their in vivo differentiated properties

PMCID: PMC2111551  PMID: 500790

Abstract

Vascular endothelial cells cultured in the presence of fibroblast growth factor (FGF) adopt at confluence a morphological appearance similar to that of the vascular endothelium in vivo. Similarly, their apical cell surface is, as in vivo, nonthrombogenic. In contrast, when the cultures are maintained in the absence of FGF, the cells undergo within two to three passages structural and functional alterations that are incompatible with their in vivo morphological appearance and physiological function. Cultures maintained in the absence of FGF no longer adopt, upon reaching confluence, the configuration of a monolayer composed of small closely apposed and nonoverlapping, cuboidal cells. Instead, confluent cultures deprived of FGF consist of large, overlapping cells which have lost the polarity of cell surface characteristic of the vascular endothelium. The apical cell surface becomes thrombogenic, as reflected by its ability to bind platelets, whereas fibronectin, which at confluence is normally associated only with the basal cell surface, can be found both on top of and underneath the cell layer. Among other changes, both sparse and confluent cultures maintained in the absence of FGF showed a greatly increased production of fibronectin. CSP-60, a cell surface protein whose appearance is correlative with the adoption of a cell monolayer configuration, can no longer be detected in cultures maintained in the absence of FGF. Overlapping endothelial cells maintained in the absence of FGF can also no longer function as a protective barrier against the uptake of ligands such as low density lipoprotein. Exposure of the culture to FGF induces a restoration of the normal endothelial characteristics concomitant with the adoption of a flattened cell monolayer morphology. These results demonstrate that, in addition to being a mitogen. FGF is involved in controlling the differentiation and phenotypic expression of the vascular endothelium. This is reflected by its effect on the morphological appearance, polarity of cell surfaces, platelet binding capacity, and barrier function of the vascular endothelium.

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Selected References

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  1. Birdwell C. R., Gospodarowicz D., Nicolson G. L. Identification, localization, and role of fibronectin in cultured bovine endothelial cells. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3273–3277. doi: 10.1073/pnas.75.7.3273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen A. B., Amrani D. L., Mosesson M. W. Heterogeneity of the cold-insoluble globulin of human plasma (CIg), a circulating cell surface protein. Biochim Biophys Acta. 1977 Aug 23;493(2):310–322. doi: 10.1016/0005-2795(77)90187-8. [DOI] [PubMed] [Google Scholar]
  3. Chen L. B., Gallimore P. H., McDougall J. K. Correlation between tumor induction and the large external transformation sensitive protein on the cell surface. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3570–3574. doi: 10.1073/pnas.73.10.3570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen L. B., Gudor R. C., Sun T. T., Chen A. B., Mosesson M. W. Control of a cell surface major glycoprotein by epidermal growth factor. Science. 1977 Aug 19;197(4305):776–778. doi: 10.1126/science.302030. [DOI] [PubMed] [Google Scholar]
  5. Engvall E., Ruoslahti E. Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int J Cancer. 1977 Jul 15;20(1):1–5. doi: 10.1002/ijc.2910200102. [DOI] [PubMed] [Google Scholar]
  6. Fielding P. E., Vlodavsky I., Gospodarowicz D., Fielding C. J. Effect of contact inhibition on the regulation of cholesterol metabolism in cultured vascular endothelial cells. J Biol Chem. 1979 Feb 10;254(3):749–755. [PubMed] [Google Scholar]
  7. Gospodarowicz D., Bialecki H., Greenburg G. Purification of the fibroblast growth factor activity from bovine brain. J Biol Chem. 1978 May 25;253(10):3736–3743. [PubMed] [Google Scholar]
  8. Gospodarowicz D., Greenburg G., Bialecki H., Zetter B. R. Factors involved in the modulation of cell proliferation in vivo and in vitro: the role of fibroblast and epidermal growth factors in the proliferative response of mammalian cells. In Vitro. 1978 Jan;14(1):85–118. doi: 10.1007/BF02618177. [DOI] [PubMed] [Google Scholar]
  9. Gospodarowicz D., Greenburg G., Birdwell C. R. Determination of cellular shape by the extracellular matrix and its correlation with the control of cellular growth. Cancer Res. 1978 Nov;38(11 Pt 2):4155–4171. [PubMed] [Google Scholar]
  10. Gospodarowicz D., Moran J. S., Braun D. L. Control of proliferation of bovine vascular endothelial cells. J Cell Physiol. 1977 Jun;91(3):377–385. doi: 10.1002/jcp.1040910307. [DOI] [PubMed] [Google Scholar]
  11. Gospodarowicz D., Moran J., Braun D., Birdwell C. Clonal growth of bovine vascular endothelial cells: fibroblast growth factor as a survival agent. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4120–4124. doi: 10.1073/pnas.73.11.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gospodarowicz D. Purification of a fibroblast growth factor from bovine pituitary. J Biol Chem. 1975 Apr 10;250(7):2515–2520. [PubMed] [Google Scholar]
  13. Gospodarowicz D., Vlodavsky I., Greenburg G., Alvarado J., Johnson L. K., Moran J. Studies on atherogenesis and corneal transplantation using cultured vascular and corneal endothelia. Recent Prog Horm Res. 1979;35:375–448. doi: 10.1016/b978-0-12-571135-7.50013-8. [DOI] [PubMed] [Google Scholar]
  14. Hedman K., Vaheri A., Wartiovaara J. External fibronectin of cultured human fibroblasts is predominantly a matrix protein. J Cell Biol. 1978 Mar;76(3):748–760. doi: 10.1083/jcb.76.3.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hynes R. O., Destree A. Extensive disulfide bonding at the mammalian cell surface. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2855–2859. doi: 10.1073/pnas.74.7.2855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jaffe E. A., Mosher D. F. Synthesis of fibronectin by cultured human endothelial cells. J Exp Med. 1978 Jun 1;147(6):1779–1791. doi: 10.1084/jem.147.6.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Levi-Montalcini R., Angeletti P. U. Nerve growth factor. Physiol Rev. 1968 Jul;48(3):534–569. doi: 10.1152/physrev.1968.48.3.534. [DOI] [PubMed] [Google Scholar]
  19. Macarak E. J., Kirby E., Kirk T., Kefalides N. A. Synthesis of cold-insoluble globulin by cultured calf endothelial cells. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2621–2625. doi: 10.1073/pnas.75.6.2621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mosher D. F., Saksela O., Vaheri A. Synthesis and secretion of alpha-2-macroglobulin by cultured adherent lung cells. Comparison with cell strains derived from other tissues. J Clin Invest. 1977 Nov;60(5):1036–1045. doi: 10.1172/JCI108854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  22. Teng N. N., Chen L. B. Thrombin-sensitive surface protein of cultured chick embryo cells. Nature. 1976 Feb 19;259(5544):578–580. doi: 10.1038/259578a0. [DOI] [PubMed] [Google Scholar]
  23. Tollefsen D. M., Feagler J. R., Majerus P. W. The binding of thrombin to the surface of human platelets. J Biol Chem. 1974 Apr 25;249(8):2646–2651. [PubMed] [Google Scholar]
  24. Vlodavsky I., Fielding P. E., Fielding C. J., Gospodarowicz D. Role of contact inhibition in the regulation of receptor-mediated uptake of low density lipoprotein in cultured vascular endothelial cells. Proc Natl Acad Sci U S A. 1978 Jan;75(1):356–360. doi: 10.1073/pnas.75.1.356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vlodavsky I., Johnson L. K., Gospodarowicz D. Appearance in confluent vascular endothelial cell monolayers of a specific cell surface protein (CSP-60) not detected in actively growing endothelial cells or in cell types growing in multiple layers. Proc Natl Acad Sci U S A. 1979 May;76(5):2306–2310. doi: 10.1073/pnas.76.5.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yamada K. M., Olden K. Fibronectins--adhesive glycoproteins of cell surface and blood. Nature. 1978 Sep 21;275(5677):179–184. doi: 10.1038/275179a0. [DOI] [PubMed] [Google Scholar]

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