Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Jun;86(12):4544–4548. doi: 10.1073/pnas.86.12.4544

An activated form of transforming growth factor beta is produced by cocultures of endothelial cells and pericytes.

A Antonelli-Orlidge 1, K B Saunders 1, S R Smith 1, P A D'Amore 1
PMCID: PMC287307  PMID: 2734305

Abstract

Using an in vitro coculture system to mimic the interactions between the cells of the vessel wall, we have previously shown that pericytes and smooth muscle cells (SMC) inhibit the growth of capillary endothelial cells (EC). We have undertaken studies to determine the mechanism of this inhibition. Using conditioned media and affinity-purified antibodies to transforming growth factor beta (TGF-beta), we now demonstrate that activated TGF-beta produced in these cocultures mediates EC growth inhibition. No inhibitory activity was detected when media conditioned by individual cultures of EC, SMC, or pericytes were examined for their effect on EC growth. In contrast, media conditioned by cocultures of EC-SMC and EC-pericytes inhibited EC proliferation to the same degree as the coculture itself. Immunoadsorption of coculture-derived conditioned media with antibodies to TGF-beta eliminated the inhibitory activity. Acid activation of serum-free media conditioned by any of the cells cultured alone yielded inhibitory activity, whereas activation of coculture conditioned media did not increase its inhibitory activity. Addition of anti-TGF-beta neutralizing antibodies to cocultures blocked the pericyte-mediated EC growth inhibition. These results indicate that latent TGF-beta is produced by these cells and it is activated by a mechanism that requires contact between the two cell types.

Full text

PDF
4544

Images in this article

4546Image
on p.4546

Selected References

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

  1. Assoian R. K., Sporn M. B. Type beta transforming growth factor in human platelets: release during platelet degranulation and action on vascular smooth muscle cells. J Cell Biol. 1986 Apr;102(4):1217–1223. doi: 10.1083/jcb.102.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baird A., Durkin T. Inhibition of endothelial cell proliferation by type beta-transforming growth factor: interactions with acidic and basic fibroblast growth factors. Biochem Biophys Res Commun. 1986 Jul 16;138(1):476–482. doi: 10.1016/0006-291x(86)90305-0. [DOI] [PubMed] [Google Scholar]
  3. Crocker D. J., Murad T. M., Geer J. C. Role of the pericyte in wound healing. An ultrastructural study. Exp Mol Pathol. 1970 Aug;13(1):51–65. doi: 10.1016/0014-4800(70)90084-5. [DOI] [PubMed] [Google Scholar]
  4. Engerman R. L., Pfaffenbach D., Davis M. D. Cell turnover of capillaries. Lab Invest. 1967 Dec;17(6):738–743. [PubMed] [Google Scholar]
  5. Folkman J., Haudenschild C. C., Zetter B. R. Long-term culture of capillary endothelial cells. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5217–5221. doi: 10.1073/pnas.76.10.5217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fryling C. M., Iwata K. K., Johnson P. A., Knott W. B., Todaro G. J. Two distinct tumor cell growth-inhibiting factors from a human rhabdomyosarcoma cell line. Cancer Res. 1985 Jun;45(6):2695–2699. [PubMed] [Google Scholar]
  7. Fràter-Schröder M., Müller G., Birchmeier W., Böhlen P. Transforming growth factor-beta inhibits endothelial cell proliferation. Biochem Biophys Res Commun. 1986 May 29;137(1):295–302. doi: 10.1016/0006-291x(86)91209-x. [DOI] [PubMed] [Google Scholar]
  8. Furchgott R. F., Zawadzki J. V. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 1980 Nov 27;288(5789):373–376. doi: 10.1038/288373a0. [DOI] [PubMed] [Google Scholar]
  9. Gitlin J. D., D'Amore P. A. Culture of retinal capillary cells using selective growth media. Microvasc Res. 1983 Jul;26(1):74–80. doi: 10.1016/0026-2862(83)90056-0. [DOI] [PubMed] [Google Scholar]
  10. Heimark R. L., Twardzik D. R., Schwartz S. M. Inhibition of endothelial regeneration by type-beta transforming growth factor from platelets. Science. 1986 Sep 5;233(4768):1078–1080. doi: 10.1126/science.3461562. [DOI] [PubMed] [Google Scholar]
  11. Herman I. M., D'Amore P. A. Microvascular pericytes contain muscle and nonmuscle actins. J Cell Biol. 1985 Jul;101(1):43–52. doi: 10.1083/jcb.101.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hobson B., Denekamp J. Endothelial proliferation in tumours and normal tissues: continuous labelling studies. Br J Cancer. 1984 Apr;49(4):405–413. doi: 10.1038/bjc.1984.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ignotz R. A., Massagué J. Type beta transforming growth factor controls the adipogenic differentiation of 3T3 fibroblasts. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8530–8534. doi: 10.1073/pnas.82.24.8530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. KUWABARA T., COGAN D. G. Retinal vascular patterns. VI. Mural cells of the retinal capillaries. Arch Ophthalmol. 1963 Apr;69:492–502. doi: 10.1001/archopht.1963.00960040498013. [DOI] [PubMed] [Google Scholar]
  15. Kalebic T., Garbisa S., Glaser B., Liotta L. A. Basement membrane collagen: degradation by migrating endothelial cells. Science. 1983 Jul 15;221(4607):281–283. doi: 10.1126/science.6190230. [DOI] [PubMed] [Google Scholar]
  16. Knabbe C., Lippman M. E., Wakefield L. M., Flanders K. C., Kasid A., Derynck R., Dickson R. B. Evidence that transforming growth factor-beta is a hormonally regulated negative growth factor in human breast cancer cells. Cell. 1987 Feb 13;48(3):417–428. doi: 10.1016/0092-8674(87)90193-0. [DOI] [PubMed] [Google Scholar]
  17. Krycève-Martinerie C., Lawrence D. A., Crochet J., Jullien P., Vigier P. Further study of beta-TGFs released by virally transformed and non-transformed cells. Int J Cancer. 1985 Apr 15;35(4):553–558. doi: 10.1002/ijc.2910350421. [DOI] [PubMed] [Google Scholar]
  18. Larson D. M., Carson M. P., Haudenschild C. C. Junctional transfer of small molecules in cultured bovine brain microvascular endothelial cells and pericytes. Microvasc Res. 1987 Sep;34(2):184–199. doi: 10.1016/0026-2862(87)90052-5. [DOI] [PubMed] [Google Scholar]
  19. Lawrence D. A., Pircher R., Jullien P. Conversion of a high molecular weight latent beta-TGF from chicken embryo fibroblasts into a low molecular weight active beta-TGF under acidic conditions. Biochem Biophys Res Commun. 1985 Dec 31;133(3):1026–1034. doi: 10.1016/0006-291x(85)91239-2. [DOI] [PubMed] [Google Scholar]
  20. Lawrence D. A., Pircher R., Krycève-Martinerie C., Jullien P. Normal embryo fibroblasts release transforming growth factors in a latent form. J Cell Physiol. 1984 Oct;121(1):184–188. doi: 10.1002/jcp.1041210123. [DOI] [PubMed] [Google Scholar]
  21. Like B., Massagué J. The antiproliferative effect of type beta transforming growth factor occurs at a level distal from receptors for growth-activating factors. J Biol Chem. 1986 Oct 15;261(29):13426–13429. [PubMed] [Google Scholar]
  22. Müller G., Behrens J., Nussbaumer U., Böhlen P., Birchmeier W. Inhibitory action of transforming growth factor beta on endothelial cells. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5600–5604. doi: 10.1073/pnas.84.16.5600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nicolson G. L. Cell surface molecules and tumor metastasis. Regulation of metastatic phenotypic diversity. Exp Cell Res. 1984 Jan;150(1):3–22. doi: 10.1016/0014-4827(84)90696-7. [DOI] [PubMed] [Google Scholar]
  24. Orlidge A., D'Amore P. A. Inhibition of capillary endothelial cell growth by pericytes and smooth muscle cells. J Cell Biol. 1987 Sep;105(3):1455–1462. doi: 10.1083/jcb.105.3.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rhodin J. A. Ultrastructure of mammalian venous capillaries, venules, and small collecting veins. J Ultrastruct Res. 1968 Dec;25(5):452–500. doi: 10.1016/s0022-5320(68)80098-x. [DOI] [PubMed] [Google Scholar]
  26. Sporn M. B., Roberts A. B., Wakefield L. M., de Crombrugghe B. Some recent advances in the chemistry and biology of transforming growth factor-beta. J Cell Biol. 1987 Sep;105(3):1039–1045. doi: 10.1083/jcb.105.3.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tilton R. G., Miller E. J., Kilo C., Williamson J. R. Pericyte form and distribution in rat retinal and uveal capillaries. Invest Ophthalmol Vis Sci. 1985 Jan;26(1):68–73. [PubMed] [Google Scholar]
  28. Voyta J. C., Via D. P., Butterfield C. E., Zetter B. R. Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J Cell Biol. 1984 Dec;99(6):2034–2040. doi: 10.1083/jcb.99.6.2034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wakefield L. M., Smith D. M., Flanders K. C., Sporn M. B. Latent transforming growth factor-beta from human platelets. A high molecular weight complex containing precursor sequences. J Biol Chem. 1988 Jun 5;263(16):7646–7654. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES