CoChat
Sign Up
Cited by 175

The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.

Isolation of endothelial cells from fresh tissues

van Beijnum, Judy R, Rousch, Mat, Castermans, Karolien, van der Linden, Edith, Griffioen, Arjan W·2008·Nature Protocols
DOI 10.1038/nprot.2008.71
Open this paper in CoChat — free account

Abstract

Here, we present a protocol for the isolation of endothelial cells (ECs) from tissues. ECs make up a minor population of cells in a tissue, but play a major role in tissue homeostasis, as well as in diverse pathologies. To understand the biology of ECs, characterization of this cell population is highly desirable, but requires the availability of purified cells. For this purpose, tissues are mechanically minced and subsequently digested enzymatically with collagenase and dispase. ECs in the resulting single-cell suspension are labeled with Abs against EC surface antigens and separated from the remainder of the cells and debris by capture with magnetic beads or by high-speed cell sorting. Purified ECs are viable and suitable for characterization of diverse cellular properties. This protocol is optimized for human tissues but can also be adapted for use with other species. Depending on the tissue, the procedure can be completed in approximately 6 h.

Overview

Integrity signals

Registry

Crossref · 10.1038/nprot.2008.71

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Verification

Claims the paper makes

This paper’s full text isn’t openly available, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Citations

Cites

Research works printed but not linked: 29

  1. Griffioen, A.W. & Molema, G. Angiogenesis: potentials for pharmacologic intervention in the treatment of cancer, cardiovascular diseases, and chronic inflammation. Pharmacol. Rev. 52, 237–268 (2000). · DOI
  2. van Beijnum, J.R. et al. Gene expression of tumor angiogenesis dissected: specific targeting of colon cancer angiogenic vasculature. Blood 108, 2339–2348 (2006). · DOI
  3. Hida, K. et al. Tumor-associated endothelial cells with cytogenetic abnormalities. Cancer Res. 64, 8249–8255 (2004). · DOI
  4. Ades, E.W. et al. HMEC-1: establishment of an immortalized human microvascular endothelial cell line. J. Invest. Dermatol. 99, 683–690 (1992). · DOI
  5. Grange, C. et al. Isolation and characterization of human breast tumor-derived endothelial cells. Oncol. Rep. 15, 381–386 (2006).
  6. Folkman, J., Haudenschild, C.C. & Zetter, B.R. Long-term culture of capillary endothelial cells. Proc. Natl. Acad. Sci. USA 76, 5217–5221 (1979). · DOI
  7. Manconi, F., Markham, R. & Fraser, I.S. Culturing endothelial cells of microvascular origin. Methods Cell Sci. 22, 89–99 (2000). · DOI
  8. Hillen, H.F., Melotte, V., van Beijnum, J.R. & Griffioen, A.W. Endothelial cell biology. In Angiogenesis Assays: A Critical Appraisal of Current Techniques (eds. Staton, C., Lewis, C. & Bicknell, R.) 1–38 (John Wiley & Sons, Chichester, West Sussex, UK, 2006).
  9. van Beijnum, J.R., van der Linden, E. & Griffioen, A.W. Angiogenic profiling and comparison of immortalized endothelial cells for functional genomics. Exp. Cell Res. 314, 264–272 (2008). · DOI
  10. Ibrahim, S.F. & van den Engh, G. High-speed cell sorting: fundamentals and recent advances. Curr. Opin. Biotechnol. 14, 5–12 (2003). · DOI
  11. Wynick, D. & Bloom, S.R. Magnetic bead separation of anterior pituitary cells. Neuroendocrinology 52, 560–565 (1990). · DOI
  12. Fritsch, G. et al. Characterization of hematopoietic stem cells. Ann. NY Acad. Sci. 770, 42–52 (1995). · DOI
  13. Garner, D.L. Flow cytometric sexing of mammalian sperm. Theriogenology 65, 943–957 (2006). · DOI
  14. Favre, C.J. et al. Expression of genes involved in vascular development and angiogenesis in endothelial cells of adult lung. Am. J. Physiol. Heart Circ. Physiol. 285, H1917–H1938 (2003). · DOI
  15. Hewett, P.W. & Murray, J.C. Immunomagnetic purification of human microvessel endothelial cells using Dynabeads coated with monoclonal antibodies to PECAM-1. Eur. J. Cell Biol. 62, 451–454 (1993).
  16. Abbot, S.E., Kaul, A., Stevens, C.R. & Blake, D.R. Isolation and culture of synovial microvascular endothelial cells. Characterization and assessment of adhesion molecule expression. Arthritis Rheum. 35, 401–416 (1992). · DOI
  17. Grimwood, J., Bicknell, R. & Rees, M.C. The isolation, characterization and culture of human decidual endothelium. Hum. Reprod. 10, 2142–2148 (1995). · DOI
  18. Madden, S.L. et al. Vascular gene expression in nonneoplastic and malignant brain. Am. J. Pathol. 165, 601–608 (2004). · DOI
  19. Parker, B.S. et al. Alterations in vascular gene expression in invasive breast carcinoma. Cancer Res. 64, 7857–7866 (2004). · DOI
  20. St. Croix, B. et al. Genes expressed in human tumor endothelium. Science 289, 1197–1202 (2000). · DOI
  21. Buckanovich, R.J. et al. Tumor vascular proteins as biomarkers in ovarian cancer. J. Clin. Oncol. 25, 852–861 (2007). · DOI
  22. Seaman, S. et al. Genes that distinguish physiological and pathological angiogenesis. Cancer Cell 11, 539–554 (2007). · DOI
  23. Lu, C. et al. Gene alterations identified by expression profiling in tumor-associated endothelial cells from invasive ovarian carcinoma. Cancer Res. 67, 1757–1768 (2007). · DOI
  24. van Beijnum, J.R. & Griffioen, A.W. In silico analysis of angiogenesis associated gene expression identifies angiogenic stage related profiles. Biochim. Biophys. Acta 1755, 121–134 (2005).
  25. Dirkx, A.E. et al. Tumor angiogenesis modulates leukocyte-vessel wall interactions in vivo by reducing endothelial adhesion molecule expression. Cancer Res. 63, 2322–2329 (2003).
  26. Griffioen, A.W. Phenotype of the tumor vasculature; cell adhesion as a target for tumor therapy. Cancer J. 10, 249–255 (1997).
  27. Baeten, C.I., Wagstaff, J., Verhoeven, I.C., Hillen, H.F. & Griffioen, A.W. Flow cytometric quantification of tumour endothelial cells; an objective alternative for microvessel density assessment. Br. J. Cancer 87, 344–347 (2002). · DOI
  28. Wang, Y. & Sheibani, N. Expression pattern of alternatively spliced PECAM-1 isoforms in hematopoietic cells and platelets. J. Cell. Biochem. 87, 424–438 (2002). · DOI
  29. Castermans, K. & Griffioen, A.W. Tumor blood vessels, a difficult hurdle for infiltrating leukocytes. Biochim. Biophys. Acta 1776, 160–174 (2007).

Cited by

Sorted by citations

See all 175

Metadata

Authors
van Beijnum, J. R., Rousch, M., Castermans, K., van der Linden, E., & Griffioen, A. W.
Venue
Nature Protocols
Publisher
Springer Science and Business Media LLC
Published
2008-06
Volume
3
Issue
6
Pages
1085-1091
ISSN
  • 1754-2189
  • 1750-2799
DOI
10.1038/nprot.2008.71
Cited by
175
References
2
Language
en
Licence
www.springer.com/tdm (tdm)

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review