Methods for the isolation, culture and characterisation of equine pulmonary artery endothelial cells.
Equine endothelial cells were isolated from the pulmonary artery by enzymatic digestion and grown to confluency. The cells were characterised by positive immunofluorescent staining for von Willebrand factor and NADPH-diaphorase staining for nitric oxide synthase. Measurements of endothelins indicated that there were significant release rates from the cells for up to six hours. Measurements of intracellular calcium concentration showed that the application of bradykinin caused a transient increase in calcium concentration with similar characteristics to those observed in other endothelial cell preparations. These tests verify the endothelial character of these cells and establish the method as a reliable means of producing a primary culture of equine endothelial cells.
MacEachern
K E
KE
University of Glasgow Veterinary School.
Smith
G L
GL
Nolan
A M
AM
D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002118:Calcium / Q000032:analysis; D002450:Cell Communication / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary*; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002478:Cells, Cultured; D016232:Endothelins / Q000032:analysis; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D005455:Fluorescent Antibody Technique / Q000379:methods / Q000662:veterinary; D006736:Horses / Q000502:physiology*; D009252:NADPH Dehydrogenase / Q000032:analysis; D018377:Neurotransmitter Agents / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000032:analysis; D009573:Nitrites / Q000032:analysis; D011651:Pulmonary Artery / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D014841:von Willebrand Factor / Q000032:analysis
D016232:Endothelins; D018377:Neurotransmitter Agents; D009573:Nitrites; D014841:von Willebrand Factor; D019001:Nitric Oxide Synthase; D009252:NADPH Dehydrogenase; D001920:Bradykinin; D002118:Calcium