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Production of immortal human endothelial cell lines by strontium phosphate transfection and electroporation of SV40 sequences

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Summary

Eleven human endothelial cell lines have been produced by introducing sequences from the DNA tumor virus SV40 into human umbilical vein endothelial cells either by strontium phosphate coprecipitation or electroporation. The resultant lines were confirmed as being endothelial in origin by their production of endothelial-specific von Willebrand factor. The growth characteristics of the different lines in normal and reduced levels of serum was determined, as was their cellular response to endothelial cell growth supplement in combination with heparin, basic fibroblast growth factor, transforming growth factor-alpha, and epidermal growth factor.

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References

  1. Booyse, F. M.; Scheinbuks, J.; Lin, P. H., et al. Isolation and interrelationships of the multiple molecular tissue-type and urokinase-type plasminogen activator forms produced by cultured human umbilical vein endothelial cells. J. Biol. Chem. 263:51529–51538; 1988.

    Google Scholar 

  2. Brash, D. E.; Reddel, R. R.; Quanrud, M., et al. Strontium phosphate transfection of human cells in primary culture: stable expression of the simian virus 40 large-T-antigen gene in primary human bronchial epithelial cells. Mol. Cell. Biol. 7:2031–2034; 1987.

    PubMed  CAS  Google Scholar 

  3. Camussi, G.; Aglietta, M.; Malavasi, F., et al. The release of plateletactivating factor from human endothelial cells in culture. J. Immunol. 131:2397–2403; 1983.

    PubMed  CAS  Google Scholar 

  4. Choi, Y.; Lee, I.; Ross, S. R. Requirement for the simian virus 40 small tumour antigen in tumorigenesis in transgenic mice. Mol. Cell. Biol. 8:3382–3390; 1988.

    PubMed  CAS  Google Scholar 

  5. Fickling, S. A.; Tooze, J. A.; Whitley, G. St. J. Characterization of human umbilical vein endothelial cell lines produced by transfection with the early region of SV40. Exp. Cell Res. 201:517–521; 1992.

    Article  PubMed  CAS  Google Scholar 

  6. Geisow, M. J. Anti-angiogenesis: towards a molecular tourniquet. TIBTECH 9:78–79; 1991.

    CAS  Google Scholar 

  7. Gimbrone, M. A.; Fareed, G. C. Transformation of cultured human vascular endothelium by SV40 DNA. Cell 9:685–693; 1976.

    Article  PubMed  Google Scholar 

  8. Gospodarowicz, D. Growth factors for vascular endothelial cells. In: Piper, H. M., ed. Cell culture techniques in heart and vessel research. Berlin, FRG: Springer Verlag Press; 1990:230–244.

    Google Scholar 

  9. Graham, F. L.; van der Eb, A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467; 1973.

    Article  PubMed  CAS  Google Scholar 

  10. Iijima, S.; Ishida, M.; Nakajima-Iijima, S., et al. Immortalization of human endothelial cells by origin-defective simian virus 40 DNA. Agric. Biol. Chem. 55:2847–2853; 1991.

    CAS  Google Scholar 

  11. 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.

    Article  PubMed  CAS  Google Scholar 

  12. Kaighn, M. E.; Reddel, R. R.; Sechner, J. F., et al. Transformation of human neonatal prostate epithelial cells by strontium phosphate transfection with a plasmid containing SV40 early region genes. Cancer Res. 49:3050–3056; 1989.

    PubMed  CAS  Google Scholar 

  13. Kreuzburg-Duffy, U.; MacDonald, C. Establishment of immortalized cell lines from mouse peritoneal macrophages following transformation with SV40 early region DNA deleted at the origin of replication. Immunology 72:368–372; 1991.

    PubMed  CAS  Google Scholar 

  14. Lassalle, P.; LaGrou, C.; Delneste, Y., et al. Human endothelial cells transfected by SV40 T antigens: characterization and potential use as a source of normal endothelial factors. Eur. J. Immunol. 22:425–431; 1992.

    Article  PubMed  CAS  Google Scholar 

  15. Lemoine, N. R.; Mayall, E. S.; Hones, T., et al. Characterization of human thyroid epithelial cells immortalisedin vitro by simian virus 40 DNA transfection. Br. J. Cancer 60:897–903; 1989.

    PubMed  CAS  Google Scholar 

  16. Levin, R. I.; Weksler, B. B.; Marcus, A. J., et al. Prostacyclin production by endothelial cells. In: Jaffe, E. A., ed. Biology of endothelial cells. Boston: Martinus Nijhoff Publishers; 1984:228–247.

    Google Scholar 

  17. Liotta, L. A.; Steeg, P. S.; Stetler-Stevenson, W. G. Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell 64:327–336; 1991.

    Article  PubMed  CAS  Google Scholar 

  18. Loskutoff, D. J.; Levin, E. Properties of plasminogen activators produced by endothelial cells. In: Jaffe, E. A., ed. Biology of endothelial cells. Boston: Martinus Nijhoff Publishers; 1984:200–208.

    Google Scholar 

  19. Maxwell, I. A.; Maxwell, F. Electroporation of mammalian cells with a firefly luciferase expression plasmid: kinetics of transient expression differ markedly among cell types. DNA (New York) 7:557–562; 1988.

    CAS  Google Scholar 

  20. Moncada, S.; Palmer, R. M. J.; Higgs, E. A. Vascular endothelium, receptors and transduction mechanisms. In: Catracas, J. D.; Gillis, C. N.; Ryan, U. S., eds. New York: Plenum Press; 1991:217–223.

    Google Scholar 

  21. Rubin, H.; Figge, J.; Bladon, M. T., et al. Role of small t antigen in the acute transforming activity of SV40. Cell 30:469–480; 1982.

    Article  PubMed  CAS  Google Scholar 

  22. Thornton, S. C.; Mueller, S. N.; Levine, E. M. Human endothelial cells: use of heparin in cloning and long-term serial cultivation. Science 222:623–625; 1983.

    Article  PubMed  CAS  Google Scholar 

  23. Van Hinsbergh, V. W. M.; Binnema, D.; Scheffer, M. A., et al. Production of plasminogen activators and inhibitor by serially propagated endothelial cells from adult human blood vessels. Arteriosclerosis 7:389–400; 1987.

    PubMed  Google Scholar 

  24. Wagner, D. D. Cell biology of von Willebrand factor. Ann. Rev. Cell. Biol. 6:217–246; 1990.

    PubMed  CAS  Google Scholar 

  25. Yanagisawa, M.; Kurihara, H.; Kimura, S., et al. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 332:411–415; 1988.

    Article  PubMed  CAS  Google Scholar 

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Fitzgerald, U., McLean, J.S. & MacDonald, C. Production of immortal human endothelial cell lines by strontium phosphate transfection and electroporation of SV40 sequences. In Vitro Cell Dev Biol - Animal 30, 236–242 (1994). https://doi.org/10.1007/BF02632045

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  • DOI: https://doi.org/10.1007/BF02632045

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