Postnatal neovascularization by endothelial progenitor cells immortalized with the simian virus 40T antigen gene.
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.
Qiu
Hui-Ying
HY
Division of Hematology and Oncology, Department of Medicine, Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan.
Fujimori
Yoshihiro
Y
Nishioka
Keisuke
K
Yamaguchi
Nobuko
N
Hashimoto-Tamaoki
Tomoko
T
Sugihara
Ayako
A
Terada
Nobuyuki
N
Nagaya
Noritoshi
N
Kanda
Munetake
M
Kobayashi
Naoya
N
Tanaka
Noriaki
N
Westerman
Karen A
KA
Leboulch
Philippe
P
Hara
Hiroshi
H
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
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