Isolation and maintenance of human pulmonary artery endothelial cells in culture isolated from transplant donors.
Even though endothelial cells from different locations have similarities, there are potential morphological and functional differences between cells from different vascular regions, as well as between species. Our laboratory is interested in studying the molecular regulation of vasoactive substances in pulmonary vasculature. Therefore, we have developed reproducible methodology to isolate and maintain cultures of human pulmonary artery endothelial cells. The major innovation has been the employment of sections of pulmonary artery from heart transplant donors, from which endothelial cells are isolated. Cell monolayers were identified as endothelial cells by phase-contrast microscopy. Representative dishes of cells were further characterized by indirect immunofluorescent staining for factor VIII antigen, uptake of acetylated low-density lipoprotein, and electron microscopy. These cells were also evaluated for the expression of endothelin-1 (ET-1), a vasoactive 21-amino acid peptide derived from endothelial cells. The cells expressed ET-1 peptide and mRNA as determined by radioimmunoassay and Northern analysis, respectively. We also demonstrated that these cells are useful in transient transfection experiments for potential evaluation of promoter elements. The availability and relevance of these cells provide an important investigative tool for studies on human pulmonary vascular disease.
Visner
G A
GA
Department of Pediatrics, University of Florida, Gainesville.
Staples
E D
ED
Chesrown
S E
SE
Block
E R
ER
Zander
D S
DS
Nick
H S
HS
D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D019332:Endothelin-1; D016232:Endothelins / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D005169:Factor VIII / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D013006:Growth Hormone / Q000235:genetics; D016027:Heart Transplantation; D006801:Humans; D051379:Mice; D008854:Microscopy, Electron; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014019:Tissue Donors; D014162:Transfection
D019332:Endothelin-1; D016232:Endothelins; D011498:Protein Precursors; D012333:RNA, Messenger; D005169:Factor VIII; D013006:Growth Hormone