Skip to main content
Log in

Endothelium specific Weibel-Palade bodies in a continuous human cell line, EA.hy926

  • Regular Papers
  • Published:
In Vitro Cellular & Developmental Biology Aims and scope Submit manuscript

Summary

Weibel-Palade bodies are ultrastructurally defined organelles found only in vascular endothelial cells. Because endothelium in corpo is very dispersed, isolation and further characterization of this organelle has been dependent on increasing the number of cells in culture. However, primary isolates of endothelial cells have a limited replication potential and tend to senesce in culture. In this report, EA.hy926, a continuously replicating cell line derived from human endothelium, is shown to contain Weibel-Palade bodies. Electron micrographs demonstrate the ultrastructural characteristics of these tissue-specific organelles and their cytoplasmic distribution in EA.hy926 cells. Von Willebrand factor, which has been shown to exist in Weibel Palade bodies, is demonstrated by immunofluorescence in discrete rod-shaped organelles whose size, shape, and distribution are consistent with that of Weibel-Palade bodies in primary endothelial cell cultures. Rapid release of von Willebrand factor can be induced by calcium ionophore, and large multimeric forms of the protein are found in EA.hy926 cells. These two properties are consistent with the function currently ascribed to Weibel Palade bodies: storage of multimerized von Willebrand factor. Thus ultrastructural, immunologic, and functional data establish the existence of this as yet poorly understood tissue-specific organelle in a continuous, vigorously replicating human cell line.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Ager, A.; Gordon, J. L.; Moncada S., et al. Effects of isolation and culture on prostaglandin synthesis by porcine aortic endothelial and smooth muscle cells. J. Cell Physiol. 110:9–16; 1982.

    Article  PubMed  CAS  Google Scholar 

  2. Beretz, A.; Freyssinet, J. M.; Gauchy, J., et al. Stability of the thrombin-thrombomodulin complex on the surface of endothelial cells from human saphenous vein or from the cell line EA.hy926. Biochem. J. 259:35–40; 1989.

    PubMed  CAS  Google Scholar 

  3. Bonfanti, R.; Furie, B. C.; Furie, B., et al. PADGEM (GMP-140) is a component of Weibel-Palade bodies of human endothelial cells. Blood 73:1109–1112; 1989.

    PubMed  CAS  Google Scholar 

  4. Bradley, L. A.; Franco, E. L.; Reisner, H. M. Use of monoclonal antibodies in an enzyme immunoassay for factor VIII-related antigen. Clin. Chem. 30:87–90; 1989.

    Google Scholar 

  5. Bussolino, F.; Biffignandi, P.; Arese, P. Platelet-activating factor—a powerful lipid autacoid possibly involved in microangiopathy. Acta Haematol (Basel) 75:129–140; 1986.

    Article  CAS  Google Scholar 

  6. Bussolino, F.; Wang, J. M.; Defilippi P., et al. Induction of human endothelial cells migration and proliferation by granulocyte- and granulocyte-macrophage-colony stimulating factor. Nature 337:471–473; 1989.

    Article  PubMed  CAS  Google Scholar 

  7. Cramer, E. M.; Meyer, D.; le Menn, R., et al. Eccentric localization of von Willebrand factor in an internal structure of platelet alpha-granule resembling that of Weibel-Palade bodies. Blood 66:710–713; 1985.

    PubMed  CAS  Google Scholar 

  8. DeBault, L. E.; Esmon, N. L.; Esmon, C. T., et al. Thrombomodulin expressed in human endothelial cell hybrid. Fed. Proc. 43:7183; 1984.

    Google Scholar 

  9. Del Vecchio, P. J.; Smith, J. R. Aging of endothelium in culture: decrease in angiotensin converting enzyme activity. Cell. Biol. Int. Rep. 6:379–384; 1982.

    Article  PubMed  Google Scholar 

  10. Edgell, C. J. S.; McDonald, C. C.; Graham, J. B. A permanent cell line expressing factor VIII related antigen established by hybridization. Proc. Natl. Acad. Sci. USA 80:3734–3737; 1983.

    Article  PubMed  CAS  Google Scholar 

  11. Emeis, J. J.; Edgell, C. J. S. Fibrinolytic properties of a human endothelial hybrid cell line (EA.hy926), Blood 71:1669–1675; 1988.

    PubMed  CAS  Google Scholar 

  12. Ewenstein, B. M.; Warhol, M. J.; Handin, R. I., et al. Composition of the von Willebrand factor storage organelle (Weibel-Palade body) isolated from cultured human umbilical vein endothelial cells. J. Cell Biol. 104:1423–1433; 1987.

    Article  PubMed  CAS  Google Scholar 

  13. Fath, K. R.; Edgell, C. J. S.; Burridge, K. Distribution of distinct integrins in focal contacts is determined by the substratum composition. J. Cell Sci. 92:67–75; 1989.

    PubMed  Google Scholar 

  14. Gimbrone, M. A. Culture of vascular endothelium. Prog. Hemostasis. Thromb. 3:1–28; 1976.

    Google Scholar 

  15. haudenschild, C. C.; Cotran, R. S.; Gimbrone, M. A., et al. Fine structure of vascular endothelium in culture. J. Ultrastruct. Res. 50:22–32; 1975.

    Article  PubMed  CAS  Google Scholar 

  16. Johnston, G. I.; Kurosky, A.; McEver, R. P. Structural and biosynthetic studies of the granule membrane protein, GMP-140, from human platelets and endothelial cells. J. Biol. Chem. 264:1816–1823; 1989.

    PubMed  CAS  Google Scholar 

  17. Johnston, G. I.; Cook, R. G.; McEver, R. P. Cloning of GMP-140, a granule membrane protein of platelets and endothelium: sequence similarity to proteins involved in cell adhesion and inflammation. Cell 56:1033–1044; 1989.

    Article  PubMed  CAS  Google Scholar 

  18. Lawrie, A. S.; Hoser, M. J.; Savidge, G. F. Phast assessment of VWF: Ag multimeric distribution. Thromb. Res. (in press); 1990.

  19. Loesberg, C.; Gonsalves, M. D.; Zandbergen, J., et al. The effect of calcium on the secretion of factor VIII-related antigen by cultured human endothelial cells. Biochim. Biophys. Acta 763:160–168; 1983.

    Article  PubMed  CAS  Google Scholar 

  20. McEver, R. P.; Beckstead, J. H.; Moore, K. L., et al. GMP-140, a platelet α-granule membrane protein, is also synthesized by vascular endothelial cells and is localized in Weibel-Palade bodies. J. Clin. Invest. 84:92–99; 1989.

    Article  PubMed  CAS  Google Scholar 

  21. Pearson, J. D.; Carleton, J. S.; Hutchings, A. Prostacyclin release stimulated by thrombin or bradykinin in porcine endothelial cells cultured from aorta and umbilical vein. Thromb. Res. 29:115–124; 1983.

    Article  PubMed  CAS  Google Scholar 

  22. Reinders, J. H.; de Groot, P. G.; Gonsalves, M. D., et al. Isolation of a storage and secretory organelle containing von Willebrand protein from cultured human endothelial cells. Biochim. Biophys. Acta 804:361–369; 1984.

    Article  PubMed  CAS  Google Scholar 

  23. Reinders, J. H.; de Groot, P. G.; Sixma, J. J., et al. Storage and secretion of von Willebrand factor by endothelial cells. Haemostasis 18:246–261; 1988.

    PubMed  CAS  Google Scholar 

  24. Rieber, A. J.; Edgell, C. J. S. Endothelium-specific gene expression. Thromb. Haemostasis 62:98; 1989.

    Google Scholar 

  25. Rosen, E. M.; Noveral, J. P.; Mueller, S. N., et al. Regulation of angiotensin I-converting enzyme activity in serially cultivated bovine endothelial cells. J. Cell Physiol. 122:30–38; 1985.

    Article  PubMed  CAS  Google Scholar 

  26. Sporn, L. A.; Marder, V. J.; Wagner, D. D. Inducible secretion of large, biologically potent von Willebrand factor multimers. Cell 46:185–190; 1986.

    Article  PubMed  CAS  Google Scholar 

  27. Suggs, J. E.; Madden, M. C.; Friedman, M., et al. Prostacyclin expression by a continuous human cell line derived from vascular endothelium. Blood 68:825–829; 1986.

    PubMed  CAS  Google Scholar 

  28. Tsai, H. M.; Nagel, R. L.; Hatcher, V. B., et al. Multimeric composition of endothelial cell-derived von Willebrand factor. Blood 73:2074–2076; 1989.

    PubMed  CAS  Google Scholar 

  29. Van Oost, B. A.; Edgell, C. J. S.; Hay, C. W., et al. Isolation of human von Willebrand factor cDNA from the EA.hy926 cell line. Biochem. Cell Biol. 64:699–705; 1986.

    Article  PubMed  Google Scholar 

  30. Wagner, D. D.; Olmsted, J. B.; Marder, V. J. Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells. J. Cell Biol. 95:355–360; 1982.

    Article  PubMed  CAS  Google Scholar 

  31. Weibel, E. R.; Palade, G. E. New cytoplasmic components in arterial endothelial. J. Cell Biol. 23:101–112; 1964.

    Article  PubMed  CAS  Google Scholar 

  32. Zimmerman, T. S.; Batlle, F. J.; Ruggeri, Z. M., et al. Multimeric analysis of factor VIII/von Willebrand factor in the diagnosis of von Willebrand's disease. In: Nakamura, R. M.; Dito, W. R.; Tucker, E. S., eds. Clinical laboratory assays: new technology and future directions. New York: Masson Publishing USA, Inc.; 1983:249–264.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Edgell, CJ.S., Haizlip, J.E., Bagnell, C.R. et al. Endothelium specific Weibel-Palade bodies in a continuous human cell line, EA.hy926. In Vitro Cell Dev Biol 26, 1167–1172 (1990). https://doi.org/10.1007/BF02623694

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02623694

Key words

Navigation