Quantitation of endothelial cell adhesiveness in vitro

Donna Lowe, Kenneth Raj*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)


One of the cardinal processes of inflammation is the infiltration of immune cells from the lumen of the blood vessel to the surrounding tissue. This occurs when endothelial cells, which line blood vessels, become adhesive to circulating immune cells such as monocytes. In vitro measurement of this adhesiveness has until now been done by quantifying the total number of monocytes that adhere to an endothelial layer either as a direct count or by indirect measurement of the fluorescence of adherent monocytes. While such measurements do indicate the average adhesiveness of the endothelial cell population, they are confounded by a number of factors, such as cell number, and do not reveal the proportion of endothelial cells that are actually adhesive. Here we describe and demonstrate a method which allows the enumeration of adhesive cells within a tested population of endothelial monolayer. Endothelial cells are grown on glass coverslips and following desired treatment are challenged with monocytes (that may be fluorescently labeled). After incubation, a rinsing procedure, involving multiple rounds of immersion and draining, the cells are fixed. Adhesive endothelial cells, which are surrounded by monocytes are readily identified and enumerated, giving an adhesion index that reveals the actual proportion of endothelial cells within the population that are adhesive.

Original languageEnglish
Article numbere52924
JournalJournal of Visualized Experiments
Issue number100
Publication statusPublished - 18 Jun 2015

Bibliographical note

Publisher Copyright:
© 2015 Creative Commons Attribution-NonCommercial License.


  • Adhesion
  • Adhesion assay
  • Atherosclerosis
  • Cellular biology
  • Endothelial cells
  • Inflammation
  • Issue 100
  • Monocytes


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