title: Postnatal neovascularization by endothelial progenitor cells immortalized with the simian virus 40T antigen gene.
abstract: Endothelial progenitor cells (EPCs) contribute to blood vessel formation in ischemic and tumorous tissues, but comprise only a small population in circulation. We attempted to immortalize putative EPCs from human cord blood. Human CD34+ cord blood cells were cultured in the presence of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF), and transfected with a retroviral vector encoding the simian virus 40 large T (SV40T) antigen. This resulted in the immortalization of cord blood cells, leading to the establishment of several cell lines. One of these lines, HYCEC-1, exhibited a phenotype characteristic of the endothelial lineage, including expression of von Willebrand factor and VEGF receptor-2 (VEGFR-2/KDR/Flk-1) and uptake of acetylated-low density lipoprotein. Flow cytometric analysis revealed that HYCEC-1 cells were strongly positive for CD31 and CD146, moderately positive for CD144, weakly positive for CD133 and CD34, and negative for CD14 and CD45. HYCEC-1 cells formed capillary-like structures on basement matrix gel in vitro. Upon transplantation into the ischemic hind limb of nude rats, HYCEC-1 cells efficiently participated in neovascularization and augmented blood flow. The immortalized HYCEC-1 cells are suggested to be a class of EPCs that can efficiently participate in postnatal neovasculogenesis in the ischemic hind limb, and may also be a useful tool for studying tumor vessel formation.
authors.affiliation: Division of Hematology and Oncology, Department of Medicine, Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan.
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Qiu
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Fujimori
authors.forename: Keisuke
authors.initials: K
authors.lastname: Nishioka
authors.forename: Nobuko
authors.initials: N
authors.lastname: Yamaguchi
authors.forename: Tomoko
authors.initials: T
authors.lastname: Hashimoto-Tamaoki
authors.forename: Ayako
authors.initials: A
authors.lastname: Sugihara
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Terada
authors.forename: Noritoshi
authors.initials: N
authors.lastname: Nagaya
authors.forename: Munetake
authors.initials: M
authors.lastname: Kanda
authors.forename: Naoya
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Noriaki
authors.initials: N
authors.lastname: Tanaka
authors.forename: Karen A
authors.initials: KA
authors.lastname: Westerman
authors.forename: Philippe
authors.initials: P
authors.lastname: Leboulch
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hara
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D001783:Blood Flow Velocity; D051929:CD146 Antigen / Q000032:analysis; D015820:Cadherins / Q000032:analysis; D002460:Cell Line; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000235:genetics / Q000276:immunology; D017690:Cell Transplantation / Q000379:methods; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006023:Glycoproteins / Q000032:analysis; D006614:Hindlimb / Q000098:blood supply / Q000601:surgery; D006801:Humans; D007511:Ischemia / Q000503:physiopathology; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic; D010455:Peptides / Q000032:analysis; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D051381:Rats; D011916:Rats, Inbred F344; D011923:Rats, Nude; D013234:Stem Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D000099152:Cadherin 5
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D000952:Antigens, Polyomavirus Transforming; D051929:CD146 Antigen; D015820:Cadherins; D016222:Fibroblast Growth Factor 2; D006023:Glycoproteins; D010455:Peptides; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A; D000099152:Cadherin 5; C000606023:PROM1 protein, human; C000606025:Prom1 protein, rat