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 language | English |
|---|---|
| Pages (from-to) | 73-77 |
| Number of pages | 5 |
| Journal | Journal of Pharmacological and Toxicological Methods |
| Volume | 32 |
| Issue number | 2 |
| DOIs | |
| State | Published - 1994 |
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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 journal › Article
}
TY - JOUR
T1 - Technique for assessment of leukocyte adherence to human umbilical vein endothelial cell monolayers
AU - McGregor, Patrick E.
AU - Agrawal, Devendra K.
AU - Edwards, John D.
PY - 1994
Y1 - 1994
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0028035319&partnerID=8YFLogxK
U2 - 10.1016/1056-8719(94)90056-6
DO - 10.1016/1056-8719(94)90056-6
M3 - Article
C2 - 7532469
AN - SCOPUS:0028035319
VL - 32
SP - 73
EP - 77
JO - Journal of Pharmacological and Toxicological Methods
JF - Journal of Pharmacological and Toxicological Methods
SN - 1056-8719
IS - 2
ER -