title: Characterization of immortalized human umbilical and iliac vein endothelial cell lines after transfection with SV40 large T-antigen.
abstract: Most in vitro studies of human endothelial cells have relied on cells derived from human umbilical veins (HUVEC); however, heterogeneity of primary cultured endothelial cells can make critical interpretation of results difficult. Several endothelial cell lines have been produced to serve as a more constant source of endothelial cells. In this study, we characterized the endothelial cell lines EVLB3 and EVLC2 derived from HUVEC, and EVLK1 and EVLK2 derived from human iliac vein endothelial cells (HIVEC). These cell lines maintained the typical endothelial cell cobblestone morphology and appeared to be growth factor independent. They lost PECAM-1 and von Willebrand factor, GP96 was reduced to the level of vascular smooth muscle cells (SMC), but aSMC-actin was far less than in vascular SMC. Antigen levels of tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor (PAI-1) were comparable with young endothelial cells, and mRNA was present for tPA, PAI-1, tissue factor (TF), tissue factor pathway inhibitor and thrombomodulin. This study revealed that mRNA and protein expression of coagulation and fibrinolytic factors was influenced by the stage of cell confluence. No differences could be detected between the endothelial cell lines derived from HUVEC and HIVEC. These cell lines may be a useful tool for studies on cellular interactions of fibrinolytic components or exploring the regulation of TF expression.
authors.affiliation: Department of Haematology, University of Groningen and University Hospital Groningen, The Netherlands. e.van.leeuwen@path.azg.nl
authors.forename: E B
authors.initials: EB
authors.lastname: van Leeuwen
authors.forename: R
authors.initials: R
authors.lastname: Veenstra
authors.forename: R
authors.initials: R
authors.lastname: van Wijk
authors.forename: G
authors.initials: G
authors.lastname: Molema
authors.forename: A
authors.initials: A
authors.lastname: Hoekstra
authors.forename: M H
authors.initials: MH
authors.lastname: Ruiters
authors.forename: J
authors.initials: J
authors.lastname: van der Meer
mesh_terms: D000952:Antigens, Polyomavirus Transforming / Q000494:pharmacology*; D015415:Biomarkers; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed / Q000737:chemistry; D019976:Cloning, Organism; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007084:Iliac Vein / Q000737:chemistry / Q000166:cytology*; D007150:Immunohistochemistry; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013925:Thromboplastin / Q000032:analysis; D010959:Tissue Plasminogen Activator / Q000032:analysis; D014162:Transfection; D014471:Umbilical Veins / Q000737:chemistry / Q000166:cytology*; D014841:von Willebrand Factor / Q000032:analysis
chemical_list: D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D017395:Plasminogen Activator Inhibitor 1; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014841:von Willebrand Factor; D013925:Thromboplastin; D010959:Tissue Plasminogen Activator