Skip to main content
SpringerLink
Log in
Book cover

Arthritis Research pp 305–321Cite as

Isolation and Analysis of Large and Small Vessel Endothelial Cells

  • Protocol

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 135))

Abstract

The in vitro isolation, propagation and study of endothelial cells (EC) is an invaluable means by which the function of the vascular endothelium in physiology and pathophysiology can be explored. In recent years heterogeneity between large and small vessel EC, between arteries and veins, and between microvascular EC derived from different organs has become increasingly apparent. This has led to the development of protocols for the isolation of these different EC. In addition, the data emerging on vascular EC function in gene-targeted mice has highlighted the need for reliable methods of isolation of murine EC. This chapter describes methods for the isolation, characterization and culture of macro- and microvascular EC from a variety of species and introduces simple approaches to investigating their surface antigen expression.

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

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Jaffe, E. A., Nachman, R. L., Becker, C. G., and Minick, C. R. (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 52(11), 2745–2756.

    Article  CAS  PubMed  Google Scholar 

  2. Jackson, C. J., Garbett, P. K., Marks, R. M., et al. (1989) Isolation and propagation of endothelial cells derived from rheumatoid synovial microvasculature. Ann. Rheum. Dis. 48(9), 733–736.

    Article  CAS  PubMed  Google Scholar 

  3. Jackson, C. J., Garbett, P. K., Nissen, B., and Schrieber, L. (1990) Binding of human endothelium to Ulex europaeus I-coated Dynabeads: application to the isolation of microvascular endothelium. J. Cell Sci. 96, 257–262.

    PubMed  Google Scholar 

  4. Abbot, S. E., Kaul, A., Stevens, C. R., and Blake, D. R. (1992) Isolation and Culture of Synovial Microvascular Endothelial Cells — Characterization and Assessment of Adhesion Molecule Expression. Arthritis Rheum. 35(4), 401–406.

    Article  CAS  PubMed  Google Scholar 

  5. Hewett, P. W., Murray, J. C., Price, E. A., Watts, M. E., and Woodcock, M. (1993) Isolation and Characterization of Microvessel Endothelial Cells from Human Mammary Adipose Tissue. In Vitro Cell Dev. Biol. Animal 29a, 325–331.

    Article  CAS  Google Scholar 

  6. Hewett, P. W. and Murray, J. C. (1993) Human Lung Microvessel Endothelial Cells — Isolation, Culture, and Characterization. Microvascular Res. 46(1), 89–102.

    Article  CAS  Google Scholar 

  7. Hewett, P. W. and Murray, J. C. (1993) Human Microvessel Endothelial Cells — Isolation, Culture and Characterization. In Vitro Cell Dev. Biol. Animal 29A(11), 823–830.

    Article  CAS  Google Scholar 

  8. Hewett, P. W. and Murray, J. C. (1993) Immunomagnetic Purification of Human Microvessel Endothelial Cells Using Dynabeads Coated with Monoclonal Antibodies to PECAM-1. Eur. J. Cell Biol. 62(2), 451–454.

    CAS  PubMed  Google Scholar 

  9. Scott, P. A. E. and Bicknell, R. (1993) The Isolation and Culture of Microvascular Endothelium. J. Cell Sci. 105, 269–273.

    PubMed  Google Scholar 

  10. Gerritsen, M. E., Shen, C. P., McHugh, M. C., et al. (1995) Activation-dependent isolation and culture of murine pulmonary microvascular endothelium. Microcirculation 2(2), 151–163.

    Article  CAS  PubMed  Google Scholar 

  11. Dong, Q. G., Bernasconi, S., Lostaglio, S., et al. (1997) A general strategy for isolation of endothelial cells from murine tissues. Characterization of two endothelial cell lines from the murine lung and subcutaneous sponge implants. Arterioscler. Thromb. Vasc. Biol. 17(8), 1599–1604.

    CAS  PubMed  Google Scholar 

  12. Lidington, E. A., Rao, R. M., Marelli-Berg, F. M., Jat, P. S., Haskard, D. O., and Mason, J. C. (2002) Conditional immortalization of growth factor-responsive cardiac endothelial cells from H-2Kb-tsA58 mice. Am. J. Physiol. Cell Physiol. 282(1), C67–C74.

    CAS  PubMed  Google Scholar 

  13. Marelli-Berg, F. M., Peek, E., Lidington, E. A., Stauss, H. J., and Lechler, R. I. (2000) Isolation of endothelial cells from murine tissue. J. Immunol. Meth. 244, 205–215.

    Article  CAS  Google Scholar 

  14. Lim, Y.-C., Garcia-Cardena, G., Allport, J. R., et al. (2003) Heterogeneity of Endothelial Cells from Different Organ Sites in T-Cell Subset Recruitment. Am. J. Pathol. 162(5), 1591–1601.

    Article  CAS  PubMed  Google Scholar 

  15. Gerritsen, M. E. (1987) Functional heterogeneity of vascular endothelial cells. Biochem. Pharmacol. 36(17), 2701–2711.

    Article  CAS  PubMed  Google Scholar 

  16. Page, C., Rose, M., Yacoub, M., and Pigott, R. (1992) Antigenic heterogeneity of vascular endothelium. Am. J. Pathol. 141(3), 673–683.

    CAS  PubMed  Google Scholar 

  17. Petzelbauer, P., Bender, J. R., Wilson, J., and Pober, J. S. (1993) Heterogeneity of Dermal Microvascular Endothelial Cell Antigen Expression and Cytokine Responsiveness In situ and in Cell Culture. J. Immunol. 151(9), 5062–5072.

    CAS  PubMed  Google Scholar 

  18. Swerlick, R. A. and Lawley, T. J. (1993) Role of Microvascular Endothelial Cells in Inflammation. J. Invest. Dermatol. 100(1), S111–S115.

    Article  Google Scholar 

  19. Mason, J. C., Yarwood, H., Tárnok, A., et al. (1996) Human Thy-1 is cytokine inducible on vascular endothelial cells and is a signaling molecule regulated by protein kinase C. J. Immunol. 157, 874–883.

    CAS  PubMed  Google Scholar 

  20. Kubota, Y., Kleinman, H. K., Martin, G. R., and Lawley, T. J. (1988) Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J. Cell Biol. 107(4), 1589–1598.

    Article  CAS  PubMed  Google Scholar 

  21. Voyta, J. C., Via, D. P., Butterfield, C. E., and Zetter, B. R. (1984) Identification and isolation of endothelial cells based on their increased uptake of acetylatedlow density lipoprotein. J. Cell Biol. 99, 2034–2040.

    Article  CAS  PubMed  Google Scholar 

  22. Wellicome, S. M., Thornhill, M. H., Pitzalis, C., et al. (1990) A monoclonal antibody that detects a novel antigen on endothelial cells that is induced by tumor necrosis factor, IL-1, or lipopolysaccharide. J. Immunol. (7), 2558–2565.

    Google Scholar 

  23. Thornhill, M. H., Wellicome, S. M., Mahiouz, D. L., Lanchbury, J. S., Kyan-Aung, U., and Haskard, D. O. (1991) Tumor necrosis factor combines with IL-4 or IFN-gamma to selectively enhance endothelial cell adhesiveness for T cells. The contribution of vascular cell adhesion molecule-1-dependent and-independent binding mechanisms. J. Immunol. 146(2), 592–598.

    CAS  PubMed  Google Scholar 

  24. Mason, J. C. and Haskard, D. O. (1994) The clinical importance of leucocyte and endothelial adhesion molecules in inflammation. Vasc. Med. Rev. 5(3), 249–275.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cardiovascular Medicine Unit, The Eric Bywaters Centre, Imperial College London, Hammersmith Hospital, London, UK

    Justin C. Mason

  2. BHF Cardiovascular Medicine Unit, The Eric Bywaters Centre, Imperial College London, Hammersmith Hospital, London, UK

    Elaine A. Lidington

  3. Department of Biology, Imperial College London, London, UK

    Helen Yarwood

Authors
  1. Justin C. Mason
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elaine A. Lidington
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Helen Yarwood
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. The Kennedy Institute of Rheumatology, Imperial College London, London, UK

    Andrew P. Cope

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Mason, J.C., Lidington, E.A., Yarwood, H. (2007). Isolation and Analysis of Large and Small Vessel Endothelial Cells. In: Cope, A.P. (eds) Arthritis Research. Methods in Molecular Medicine, vol 135. Humana Press. https://doi.org/10.1007/978-1-59745-401-8_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-401-8_19

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-344-2

  • Online ISBN: 978-1-59745-401-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation