Spontaneous transformation and immortalization of human endothelial cells |
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Authors: | Kimiko Takahashi Yoshio Sawasaki Jun-Ichi Hata Kiyoshi Mukai Tamotsu Goto |
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Institution: | (1) Department of Biochemistry, National Defense Medical College, Tokorozawa, Saitama, Japan;(2) Department of Pathophysiology, National Children's Medical Research Center, Tokyo, Japan;(3) Pathology Division, National Cancer Institute, Tokyo, Japan |
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Abstract: | Summary A new cell line from the human umbilical vein has been established and maintained for more than 5 yr (180 generations; 900
population doublings). This strain, designated ECV304, is characterized by a cobblestone monolayer growth pattern, high proliferative
potential without any specific growth factor requirement, and anchorage dependency with contact inhibition. Karyotype analysis
of this cell line reveals it to be of human chromosomal constitution with a high trisomic karyotype (mode 80). Ultrastructurally,
endothelium-specific Weibel-Palade bodies were identified. Although one of the endothelial cell markers, Factor VIII-related
antigen (VIIIR:Ag) was negative in this cell line, immunocytochemical staining for the lectin Ulex europaeus I (UEA-I), and
PHM5 (anti-human endothelium as well as glomerular epithelium monoclonal antibody) was positive, and angiotensin-converting
enzyme (ACE) activity was also demonstrated. In addition, ECV304 displayed negativity for alkaline and acid phosphatase and
for the epithelial marker keratin. All of these findings suggest that ECV304 cells originated from umbilical vein endothelial
cells by spontaneous transformation. Ultrastructurally, no viruslike particles have been detected intracellularly. Nude mouse
tumorigenicity and rabbit cornea tests were both positive. This is a report on a novel case of phenotypic alteration of normal
venous endothelial cells of human origin in vitro, and generation of a transformant with indefinite life spans. This line
may be useful in studies of some physiologically active factors available for medical use. |
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Keywords: | endothelial cells spontaneous transformation Weibel-Palade body immortalization |
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