Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1974 Mar 1;60(3):673–684. doi: 10.1083/jcb.60.3.673

HUMAN VASCULAR ENDOTHELIAL CELLS IN CULTURE

Growth and DNA Synthesis

Michael A Gimbrone Jr 1, Ramzi S Cotran 1, Judah Folkman 1
PMCID: PMC2109230  PMID: 4363161

Abstract

Human endothelial cells, obtained by collagenase treatment of term umbilical cord veins, were cultured using Medium 199 supplemented with 20% fetal calf serum. Small clusters of cells initially spread on plastic or glass, coalesced and grew to form confluent monolayers of polygonal cells by 7 days. Cells in primary and subcultures were identified as endothelium by the presence of Weibel-Palade bodies by electron microscopy. A morphologically distinct subpopulation of cells contaminating some primary endothelial cultures was selectively subcultured, and identified by ultrastructural criteria as vascular smooth muscle. Autoradiography of endothelial cells after exposure to [3H]thymidine showed progressive increases in labeling in growing cultures beginning at 24 h. In recently confluent cultures, labeling indices were 2.4% in central closely packed regions, and 53.2% in peripheral growing regions. 3 days after confluence, labeling was uniform, being 3.5 and 3.9% in central and peripheral areas, respectively. When small areas of confluent cultures were experimentally "denuded," there were localized increases in [3H]thymidine labeling and eventual reconstitution of the monolayer. Liquid scintillation measurements of [3H]thymidine incorporation in primary and secondary endothelial cultures in microwell trays showed a similar correlation of DNA synthesis with cell density. These data indicate that endothelial cell cultures may provide a useful in vitro model for studying pathophysiologic factors in endothelial regeneration.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Barile M. F., Kern J. Isolation of Mycoplasma arginini from commercial bovine sera and its implication in contaminated cell cultures. Proc Soc Exp Biol Med. 1971 Nov;138(2):432–437. doi: 10.3181/00379727-138-35913. [DOI] [PubMed] [Google Scholar]
  2. Cavallo T., Sade R., Folkman J., Cotran R. S. Tumor angiogenesis. Rapid induction of endothelial mitoses demonstrated by autoradiography. J Cell Biol. 1972 Aug;54(2):408–420. doi: 10.1083/jcb.54.2.408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Comings D. E., Okada T. A. Electron microscopy of human fibroblasts in tissue culture during logarithmic and confluent stages of growth. Exp Cell Res. 1970 Aug;61(2):295–301. doi: 10.1016/0014-4827(70)90451-9. [DOI] [PubMed] [Google Scholar]
  4. Fallon J. T., Stehbens W. E. Venous endothelium of experimental arteriovenous fistulas in rabbits. Circ Res. 1972 Oct;31(4):546–556. doi: 10.1161/01.res.31.4.546. [DOI] [PubMed] [Google Scholar]
  5. Folkman J., Merler E., Abernathy C., Williams G. Isolation of a tumor factor responsible for angiogenesis. J Exp Med. 1971 Feb 1;133(2):275–288. doi: 10.1084/jem.133.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fryer D. G., Birnbaum G., Luttrell C. N. Human endothelium in cell culture. J Atheroscler Res. 1966 Mar-Apr;6(2):151–163. doi: 10.1016/s0368-1319(66)80019-4. [DOI] [PubMed] [Google Scholar]
  7. Gaynor E. Increased mitotic activity in rabbit endothelium after endotoxin. An autoradiographic study. Lab Invest. 1971 Apr;24(4):318–320. [PubMed] [Google Scholar]
  8. Gimbrone M. A., Jr, Cotran R. S., Folkman J. Endothelial regeneration: studies with human endothelial cells in culture. Ser Haematol. 1973;6(4):453–455. [PubMed] [Google Scholar]
  9. 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]
  10. Lemeunier A., Burri P. H., Weibel E. R. Absence of acid phosphatase activity in specific endothelial organelles. Histochemie. 1969;20(2):143–149. doi: 10.1007/BF00268708. [DOI] [PubMed] [Google Scholar]
  11. Lewis L. J., Hoak J. C., Maca R. D., Fry G. L. Replication of human endothelial cells in culture. Science. 1973 Aug 3;181(4098):453–454. doi: 10.1126/science.181.4098.453. [DOI] [PubMed] [Google Scholar]
  12. MARUYAMA Y. The human endothelial cell in tissue culture. Z Zellforsch Mikrosk Anat. 1963;60:69–79. doi: 10.1007/BF00329383. [DOI] [PubMed] [Google Scholar]
  13. Martz E., Steinberg M. S. The role of cell-cell contact in "contact" inhibition of cell division: a review and new evidence. J Cell Physiol. 1972 Apr;79(2):189–210. doi: 10.1002/jcp.1040790205. [DOI] [PubMed] [Google Scholar]
  14. POMERAT C. M., SLICK W. C. Isolation and growth of endothelial cells in tissue culture. Nature. 1963 Jun 1;198:859–861. doi: 10.1038/198859a0. [DOI] [PubMed] [Google Scholar]
  15. POOLE J. C., SANDERS A. G., FLOREY H. W. The regeneration of aortic endothelium. J Pathol Bacteriol. 1958 Jan;75(1):133–143. doi: 10.1002/path.1700750116. [DOI] [PubMed] [Google Scholar]
  16. Parry E. W., Abramovich D. R. The ultrastructure of human umbilical vessel endothelium from early pregnancy to full term. J Anat. 1972 Jan;111(Pt 1):29–42. [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Sade R. M., Folkman J., Cotran R. S. DNA synthesis in endothelium of aortic segments in vitro. Exp Cell Res. 1972 Oct;74(2):297–306. doi: 10.1016/0014-4827(72)90380-1. [DOI] [PubMed] [Google Scholar]
  20. Schwartz S. M., Benditt E. P. Cell replication in the aortic endothelium: a new method for study of the problem. Lab Invest. 1973 Jun;28(6):699–707. [PubMed] [Google Scholar]
  21. Spaet T. H., Lejnieks I. Mitotic activity of rabbit blood vessels. Proc Soc Exp Biol Med. 1967 Aug-Sep;125(4):1197–1201. doi: 10.3181/00379727-125-32312. [DOI] [PubMed] [Google Scholar]
  22. Tannock I. F., Hayashi S. The proliferation of capillary endothelial cells. Cancer Res. 1972 Jan;32(1):77–82. [PubMed] [Google Scholar]
  23. WEIBEL E. R., PALADE G. E. NEW CYTOPLASMIC COMPONENTS IN ARTERIAL ENDOTHELIA. J Cell Biol. 1964 Oct;23:101–112. doi: 10.1083/jcb.23.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wright H. P. Mitosis patterns in aortic endothelium. Atherosclerosis. 1972 Jan-Feb;15(1):93–100. doi: 10.1016/0021-9150(72)90042-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES