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. 1980 Feb 1;84(2):281–293. doi: 10.1083/jcb.84.2.281

Anchorage independent growth and plasminogen activator production by bovine endothelial cells

PMCID: PMC2110557  PMID: 6991508

Abstract

Endothelial cells obtained from the aortae of 1- to 2-d-old calves were cloned at high efficiency using fibrin-coated dishes. Primary cultures as well as clones derived from them produced high fibrinolytic activity when grown on 125I-fibrin-coated dishes which was 90% dependent upon the presence of plasminogen. High plasminogen-dependent proteolytic activity was also demonstrated in endothelial cell lysates and in the culture medium of the cells. The production and secretion of the plasminogen activator(s) were found to increase during the log phase of cell growth and to reach a maximum level at confluence. These endothelial cells exhibited morphological phenotypes comparable to those of transformed cells when grown in the presence of acid-treated fetal calf, dog, or human serum. Furthermore, they demonstrated anchorage independent growth, and large colonies were formed in semisolid media. Spontaneous neoplastic transformation of these cells was excluded by karyotypic analysis, lack of tumorigenicity in athymic nude mice, and limited lifespan in culture. Cell clones isolated from colonies grown in agarose demonstrated the same growth characteristics and proteolytic activity as before plating in agarose. High fibrinolytic activity, morphological changes in the appropriate serum, and growth in semisolid media may therefore be indicative of the migratory and/or invasive capacity of both nontransformed endothelial cells as well as tumor cells.

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Selected References

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

  1. ASTRUP T. Fibrinolysis in the organism. Blood. 1956 Sep;11(9):781–806. [PubMed] [Google Scholar]
  2. Aoki N., Von Kaulla K. N. The extraction of vascular plasminogen activator from human cadavers and a description of some of its properties. Am J Clin Pathol. 1971 Feb;55(2):171–179. doi: 10.1093/ajcp/55.2.171. [DOI] [PubMed] [Google Scholar]
  3. Beers W. H., Strickland S., Reich E. Ovarian plasminogen activator: relationship to ovulation and hormonal regulation. Cell. 1975 Nov;6(3):387–394. doi: 10.1016/0092-8674(75)90188-9. [DOI] [PubMed] [Google Scholar]
  4. Booyse F. M., Sedlak B. J., Rafelson M. E., Jr Culture of arterial endothelial cells: characterization and growth of bovine aortic cells. Thromb Diath Haemorrh. 1975 Dec 15;34(3):825–839. [PubMed] [Google Scholar]
  5. Bosmann H. B. Release of specific protease during mitotic cycle of L5178Y murine leukaemic cells by sublethal autolysis. Nature. 1974 May 10;249(453):144–145. doi: 10.1038/249144a0. [DOI] [PubMed] [Google Scholar]
  6. Bradley T. R., Metcalf D. The growth of mouse bone marrow cells in vitro. Aust J Exp Biol Med Sci. 1966 Jun;44(3):287–299. doi: 10.1038/icb.1966.28. [DOI] [PubMed] [Google Scholar]
  7. Buonassisi V., Venter J. C. Hormone and neurotransmitter receptors in an established vascular endothelial cell line. Proc Natl Acad Sci U S A. 1976 May;73(5):1612–1616. doi: 10.1073/pnas.73.5.1612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chou I. N., Black P. H., Roblin R. O. Suppression of fibrinolysin T activity fails to restore density-dependent growth inhibition to SV3T3 cells. Nature. 1974 Aug 30;250(5469):739–741. doi: 10.1038/250739a0. [DOI] [PubMed] [Google Scholar]
  9. Deutsch D. G., Mertz E. T. Plasminogen: purification from human plasma by affinity chromatography. Science. 1970 Dec 4;170(3962):1095–1096. doi: 10.1126/science.170.3962.1095. [DOI] [PubMed] [Google Scholar]
  10. Dosne A. M., Dupuy E., Bodevin E. Production of a fibrinolytic inhibitor by cultured endothelial cells derived from human umbilical vein. Thromb Res. 1978 Mar;12(3):377–387. doi: 10.1016/0049-3848(78)90309-2. [DOI] [PubMed] [Google Scholar]
  11. Eisenstein R., Kuettner K. E., Neapolitan C., Soble L. W., Sorgente N. The resistance of certain tissues to invasion. III. Cartilage extracts inhibit the growth of fibroblasts and endothelial cells in culture. Am J Pathol. 1975 Nov;81(2):337–348. [PMC free article] [PubMed] [Google Scholar]
  12. Gimbrone M. A., Jr, Fareed G. C. Transformation of cultured human vascular endothelium by SV40 DNA. Cell. 1976 Dec;9(4 Pt 2):685–693. doi: 10.1016/0092-8674(76)90132-x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Granelli-Piperno A., Vassalli J. D., Reich E. Secretion of plasminogen activator by human polymorphonuclear leukocytes. Modulation by glucocorticoids and other effectors. J Exp Med. 1977 Dec 1;146(6):1693–1706. doi: 10.1084/jem.146.6.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Horwitz A. L., Dorfman A. The growth of cartilage cells in soft agar and liquid suspension. J Cell Biol. 1970 May;45(2):434–438. doi: 10.1083/jcb.45.2.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jones P. A., Laug W. E., Benedict W. F. Fibrinolytic activity in a human fibrosarcoma cell line and evidence for the induction of plasminogen activator secretion during tumor formation. Cell. 1975 Oct;6(2):245–252. doi: 10.1016/0092-8674(75)90015-x. [DOI] [PubMed] [Google Scholar]
  18. Jones P. A., Laug W. E., Gardner A., Nye C. A., Fink L. M., Benedict W. F. In vitro correlates of transformation in C3H/10T1/2 clone 8 mouse cells. Cancer Res. 1976 Aug;36(8):2863–2867. [PubMed] [Google Scholar]
  19. 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]
  20. Laug W. E., Jones P. A., Benedict W. F. Relationship between fibrinolysis of cultured cells and malignancy. J Natl Cancer Inst. 1975 Jan;54(1):173–179. doi: 10.1093/jnci/54.1.173. [DOI] [PubMed] [Google Scholar]
  21. Loskutoff D. J., Edgington T. E. Synthesis of a fibrinolytic activator and inhibitor by endothelial cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3903–3907. doi: 10.1073/pnas.74.9.3903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MACPHERSON I., MONTAGNIER L. AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS. Virology. 1964 Jun;23:291–294. doi: 10.1016/0042-6822(64)90301-0. [DOI] [PubMed] [Google Scholar]
  23. MOSESSON M. W. The preparation of human fibrinogen free of plasminogen. Biochim Biophys Acta. 1962 Feb 26;57:204–213. doi: 10.1016/0006-3002(62)91112-5. [DOI] [PubMed] [Google Scholar]
  24. Ossowski L., Quigley J. P., Kellerman G. M., Reich E. Fibrinolysis associated with oncogenic transformation. Requirement of plasminogen for correlated changes in cellular morphology, colony formation in agar, and cell migration. J Exp Med. 1973 Nov 1;138(5):1056–1064. doi: 10.1084/jem.138.5.1056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ossowski L., Quigley J. P., Reich E. Fibrinolysis associated with oncogenic transformation. Morphological correlates. J Biol Chem. 1974 Jul 10;249(13):4312–4320. [PubMed] [Google Scholar]
  26. Paul B., Porter I. H. Giemsa banding in an established line of human malignant meningioma. Humangenetik. 1973 Apr 16;18(2):185–187. doi: 10.1007/BF00291489. [DOI] [PubMed] [Google Scholar]
  27. Pollack R., Risser R., Conlon S., Rifkin D. Plasminogen activator production accompanies loss of anchorage regulation in transformation of primary rat embryo cells by simian virus 40. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4792–4796. doi: 10.1073/pnas.71.12.4792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Quigley J. P., Ossowski L., Reich E. Plasminogen, the serum proenzyme activated by factors from cells transformed by oncogenic viruses. J Biol Chem. 1974 Jul 10;249(13):4306–4311. [PubMed] [Google Scholar]
  29. Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Russell W. C., Newman C., Williamson D. H. A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature. 1975 Feb 6;253(5491):461–462. doi: 10.1038/253461a0. [DOI] [PubMed] [Google Scholar]
  31. SCHOEFL G. I. STUDIES ON INFLAMMATION. III. GROWING CAPILLARIES: THEIR STRUCTURE AND PERMEABILITY. Virchows Arch Pathol Anat Physiol Klin Med. 1963 Nov 8;337:97–141. [PubMed] [Google Scholar]
  32. Seabright M. A rapid banding technique for human chromosomes. Lancet. 1971 Oct 30;2(7731):971–972. doi: 10.1016/s0140-6736(71)90287-x. [DOI] [PubMed] [Google Scholar]
  33. Sherman M. I., Strickland S., Reich E. Differentiation of early mouse embryonic and teratocarcinoma cells in vitro: plasminogen activator production. Cancer Res. 1976 Nov;36(11 Pt 2):4208–4216. [PubMed] [Google Scholar]
  34. TODD A. S. The histological localisation of fibrinolysin activator. J Pathol Bacteriol. 1959 Jul;78:281–283. doi: 10.1002/path.1700780131. [DOI] [PubMed] [Google Scholar]
  35. Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zetter B. R., Chen L. B., Buchanan J. M. Effects of protease treatment on growth, morphology, adhesion, and cell surface proteins of secondary chick embryo fibroblasts. Cell. 1976 Mar;7(3):407–412. doi: 10.1016/0092-8674(76)90170-7. [DOI] [PubMed] [Google Scholar]

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