Technique for assessment of leukocyte adherence to human umbilical vein endothelial cell monolayers

Patrick E. McGregor, Devendra K. Agrawal, John D. Edwards

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The interaction between endothelial cells and immuno/inflammatory cells plays an important role in the pathogenesis of vascular diseases. Inflammatory cells also activate endothelial cells and release both proliferative and cytotoxic mediators. In order to examine the interaction between leukocytes and endothelial cells and the effect of various drugs, we established the methodology for isolating and culturing the endothelial cells from human umbilical vein. Endothelial cells were harvested by using 0.1% collagenase within 48 hr of collecting the cord. Cells were grown to confluency in 96-well plates in Medium 199 containing 20% fetal calf serum, endothelial cell growth supplement, heparin, and antibiotics. Using this method, we obtained a confluent layer of the cells in all the 96 wells within 48 hr. We then examined the effect of peptides, endothelin-1, substance P, and neurokinin-A on the adherence of human blood neutrophils (purity and viability > 98%) to endothelial monolayers. All the peptides enhanced (p <0.05) the adherence of neutrophils to endothelial cells in a time-dependent manner. This method of endothelial cell culturing is reliable, reproducible, and effective in evaluating the role of various mediators and drugs on the adherence of various white blood cells to endothelium.

Original languageEnglish
Pages (from-to)73-77
Number of pages5
JournalJournal of Pharmacological and Toxicological Methods
Volume32
Issue number2
DOIs
StatePublished - 1994

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Endothelial cells
Human Umbilical Vein Endothelial Cells
Monolayers
Leukocytes
Endothelial Cells
Neutrophils
Blood
Neurokinin A
Peptides
Endothelin-1
Collagenases
Substance P
Vascular Diseases
Pharmaceutical Preparations
Cell growth
Endothelium
Heparin
Anti-Bacterial Agents
Cells
Growth

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Toxicology

Cite this

Technique for assessment of leukocyte adherence to human umbilical vein endothelial cell monolayers. / McGregor, Patrick E.; Agrawal, Devendra K.; Edwards, John D.

In: Journal of Pharmacological and Toxicological Methods, Vol. 32, No. 2, 1994, p. 73-77.

Research output: Contribution to journalArticle

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