title: Human vascular endothelial cells with extended life spans: in vitro cell response, protein expression, and angiogenesis.
abstract: An in vitro angiogenesis system was designed for screening angiogenic agonists and antagonists. In order to obtain large quantities of cells and reproducibility, human endothelial cells with extended life spans were developed by retroviral transfection. The resulting cells grown in a serum-free medium containing endothelial cell growth supplement (ECGS) have a telomerase activity, extended life spans of at least 21 passages, and an endothelial cell phenotype (diI-acetylated-LDL upake, factor VIII-related antigen, VEGFR-1 and R-2, and tissue-type plasminogen activator (tPA)) that resembled that of unaltered primary endothelial cells. Exceptions were (i) a higher expression of tPA, and (ii) a non-significant growth response to FGF-2 or VEGF stimulation. Within three-dimensional fibrin gels, specific cell clones rapidly formed tubular structures in a more reproducible manner than those observed with low-passage primary cells. Tube formation by primary endothelial cells and those with extended life spans was dependent upon FGF-2 and ECGS, respectively. Both cell types produced FGF-2 and VEGF cytokines. Increasing doses of suramin significantly decreased the size of microvessels formed by both cell lines. These functional results indicate that a vascular matrix system containing human cells with extended life spans can be successfully utilized as an in vitro assay for antiangiogenic compounds.
authors.affiliation: Oncology and Molecular Endocrinology Research Center, CHUL, Québec, Canada.
authors.forename: Edith
authors.initials: E
authors.lastname: Gagnon
authors.forename: Paola
authors.initials: P
authors.lastname: Cattaruzzi
authors.forename: May
authors.initials: M
authors.lastname: Griffith
authors.forename: Lea
authors.initials: L
authors.lastname: Muzakare
authors.forename: Katell
authors.initials: K
authors.lastname: LeFlao
authors.forename: Robert
authors.initials: R
authors.lastname: Faure
authors.forename: Richard
authors.initials: R
authors.lastname: Béliveau
authors.forename: Sabah N
authors.initials: SN
authors.lastname: Hussain
authors.forename: Michael
authors.initials: M
authors.lastname: Koutsilieris
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Doillon
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D002460:Cell Line; D016895:Culture Media, Serum-Free; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000502:physiology*; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*
chemical_list: D020533:Angiogenesis Inhibitors; D016895:Culture Media, Serum-Free; D011506:Proteins