ISOLATION AND MAINTENANCE OF HUMAN PULMONARY-ARTERY ENDOTHELIAL-CELLSIN CULTURE ISOLATED FROM TRANSPLANT DONORS

Even though endothelial cells from different locations have similariti es, there are potential morphological and functional differences betwe en cells from different vascular regions, as well as between species. Our laboratory is interested in studying the molecular regulation of v asoactive substances in pulmonary vasculature. Therefore, we have deve loped reproducible methodology to isolate and maintain cultures of hum an pulmonary artery endothelial cells. The major innovation has been t he employment of sections of pulmonary artery from heart transplant do nors, from which endothelial cells are isolated. Cell monolayers were identified as endothelial cells by phase-contrast microscopy. Represen tative dishes of cells were further characterized by indirect immunofl uorescent staining for factor VIII antigen, uptake of acetylated low-d ensity lipoprotein, and electron microscopy. These cells were also eva luated for the expression of endothelin-1 (ET-1), a vasoactive 21-amin o acid peptide derived from endothelial cells. The cells expressed ET- 1 peptide and mRNA as determined by radioimmunoassay and Northern anal ysis, respectively. We also demonstrated that these cells are useful i n transient transfection experiments for potential evaluation of promo ter elements. The availability and relevance of these cells provide an important investigative tool for studies on human pulmonary vascular disease.