Skip to main content
SpringerLink
Log in

Variability in messenger RNA levels in human umbilical vein endothelial cells of different lineage and time in culture

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

Summary

Levels of seven messenger RNA species were compared in human umbilical vein endothelial cells of different lineage and time in culture. Specifically, cells obtained from the American Type Culture Collection (ATCC) and subcultured were compared to early passage cells from cultures produced in our laboratory. Messenger RNA for tissue plasminogen activator, plaminogen activator inhibitor 1, urokinase, and thrombomodulin were expressed at higher levels in the ATCC cells. Thrombospondin, von Willebrand's Factor, and protein S messenger RNA were expressed at higher levels in the cells that we isolated. In addition, in the ATCC cells a shift in the proportion of plasminogen activator inhibitor messenger RNA from the 3.4 to the 2.4 kilobase species was found. We conclude that specific messenger RNA levels can vary considerably between cultured human umbilical vein endothelial cells. The large variation in mRNA levels which we describe has important implications for experiments involving gene expression in cultured endothelium.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Auerbach, R.; Alby, L.; Morrisey, L. W., et al. Expression of organ specific antigens on capillary endothelial cells. Microvasc. Res. 29:401–411; 1985.

    Article  PubMed  CAS  Google Scholar 

  2. Ausubel, F. F.; Brent, R.; Kingston, R. E., et al. eds. Current protocols in molecular biology. New York: John Wiley & Sons; 1987.

    Google Scholar 

  3. Barrett, T. B.; Gajdusek, C. M.; Schwartz, S. M., et al. Expression of the sis gene by endothelial cells in culture and in vivo. Proc. Natl. Acad. Sci. USA 81:6772–6774; 1984.

    Article  PubMed  CAS  Google Scholar 

  4. Booyse, F. M.; Osikowicz, G.; Feder, S., et al. Isolation and characterization of a urokinase-type plasminogen activator (M=54 000) from cultured human endothelial cells indistinguishable from urinary urokinase. J. Biol. Chem. 259:7198–7205; 1984.

    PubMed  CAS  Google Scholar 

  5. Chirgwin, J. M.; Przybyla, A. E.; MacDonald, R. F., et al. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. J. Biochem. 18:5294–5299; 1979.

    Article  CAS  Google Scholar 

  6. Collins, T.; Ginsburg, D.; Boss, J. M., et al. Cultured human endothelial cells express platelet derived growth factor B chain: cDNA cloning and structural analysis. Nature 316:748–750; 1985.

    Article  PubMed  CAS  Google Scholar 

  7. Collins, T.; Pober, J. S.; Gimbrone, M. A., et al. Cultured human endothelial cells express platelet derived growth factor A chain. Am. J. Pathol. 126:7–12; 1987.

    PubMed  CAS  Google Scholar 

  8. Fisher, R.; Waller, E. K.; Grossi, G., et al. Isolation and characterization of the human tissue-type plasminogen activator structural gene including its 5′ flanking region. J. Biol. Chem. 260:11223–11230; 1985.

    PubMed  CAS  Google Scholar 

  9. Ginsburg, D.; Handin, R. I.; Bonthron, D. T., et al. Human von Willebrand factor (VWF): isolation of complementary DNA (cDNA) clones and chromosomal localization. Science 228:1401–1406; 1985.

    Article  PubMed  CAS  Google Scholar 

  10. Ginsburg, D.; Zeheb, R.; Yang, A. Y., et al. cDNA cloning of human plasminogen activator inhibitor from endothelial cells. J. Clin. Invest. 78:1673–1680; 1986.

    PubMed  CAS  Google Scholar 

  11. Goldsmith, J. C.; McCormick, J. J.; Yen, A. Endothelial cell cycle kinetics. Lab. Invest. 51:643–677; 1984.

    PubMed  CAS  Google Scholar 

  12. Gumkowski, F.; Kaminska, G.; Kaminski, M., et al. Heterogeneity of mouse vascular endothelium. Blood Vessels 24:11–23; 1987.

    PubMed  CAS  Google Scholar 

  13. Jaffe, E. A.; Nachman, R. L.; Becker, C. G., et al. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 52:2745–2756; 1973.

    PubMed  CAS  Google Scholar 

  14. Lawler, J.; Hynes, R. O. The structure of human thrombospondin, an adhesive glycoprotein with multiple calciumbinding sites and homologies with several different proteins. J. Cell. Biol. 103:1635–1648; 1986.

    Article  PubMed  CAS  Google Scholar 

  15. Levin, E. G.; Loskutoff, D. J. Comparative studies of the fibrinolytic activity of cultured vascular cells. Thromb. Res. 15:869–878; 1979.

    Article  PubMed  CAS  Google Scholar 

  16. Levin, E. G.; Loskutoff, D. J. Regulation of plasminogen activator production by cultured endothelial cells. Ann. NY Acad. Sci. 401:184–194; 1982.

    Article  PubMed  CAS  Google Scholar 

  17. Levin, E. G. Latent tissue plasminogen activator produced by human endothelial cells in culture: evidence for an enzyme-inhibitor complex. Proc. Natl. Acad. Sci. USA 80:6804–6808; 1983.

    Article  PubMed  CAS  Google Scholar 

  18. Lundwall, A.; Dackowski, W.; Cohen, E., et al. Isolation and sequence of the cDNA for human protein S, a regulator of blood coagulation. Proc. Natl. Acad. Sci. USA 83:6716–6720; 1986.

    Article  PubMed  CAS  Google Scholar 

  19. Mueller, S.; Rosen, E. M.; Levine, E. M. Cellular senescence in a cloned strain of bovine fetal aortic endothelial cells. Science 207:889–891; 1980.

    Article  PubMed  CAS  Google Scholar 

  20. Ody, C.; Seillan, C.; Russo-Marie, F. 6-Keto-prostaglandin-Flα, prostaglandins F2, F2α, and thromboxane B2 production by endothelial cells, smooth muscle cells, and fibroblasts cultured from piglet aorta. Biochim. Biophys. Acta. 712:103–110; 1982.

    PubMed  CAS  Google Scholar 

  21. Ross, R. The pathogenesis of atherosclerosis—an update. N. Engl. J. Med. 314:488–500; 1986.

    Article  PubMed  CAS  Google Scholar 

  22. Sawdey, M.; Ny, T.; Loskutoff, D. J. Messenger RNA for plasminogen activator inhibitor. Thromb. Res. 41:151–160; 1986.

    Article  PubMed  CAS  Google Scholar 

  23. Schleef, R. R.; Bevilacqua, M. P.; Sawdey, M., et al. Cytokine activation of vascular endothelium. J. Biol. Chem. 263:5797–5803; 1988.

    PubMed  CAS  Google Scholar 

  24. Sieff, C. A.; Tsai, S.; Faller, D. F. Interleukin 1 induces human endothelial cell production of granulocyte-macrophage colony stimulating factor. J. Clin. Invest. 79:48–51; 1987.

    Article  PubMed  CAS  Google Scholar 

  25. Speiser, W.; Anders, E.; Preissner, K. T., et al. Differences in coagulant and fibrinolytic activities of cultured human endothelial cells derived omental tissue microvessels and umbilical veins. Blood 69:964–967; 1987.

    PubMed  CAS  Google Scholar 

  26. Van Zonneveld, A. J.; Chang, G. T. G.; van den Berg, J., et al. Quantification of tissue-type plasminogen activator (t-PA) mRNA in human endothelial cell culture by hybridization with a t-PA cDNA probe. Biochem. J. 235:385–390; 1986.

    PubMed  Google Scholar 

  27. Wautier, M. P.; Wautier, J. L. Human endothelial cell modifications during long term culture. Thromb. Heamost. 58:151; 1987 (Abstr).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cardiac Unit, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Jackson 12, 02114, Boston, Massachusetts

    David Dichek & Thomas Quertermous

Authors
  1. David Dichek
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Thomas Quertermous
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dichek, D., Quertermous, T. Variability in messenger RNA levels in human umbilical vein endothelial cells of different lineage and time in culture. In Vitro Cell Dev Biol 25, 289–292 (1989). https://doi.org/10.1007/BF02628468

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02628468

Key words

Search

Navigation