Generation of human pulmonary microvascular endothelial cell lines

Details

Serval ID
serval:BIB_48BF8B2E3588
Type
Article: article from journal or magazin.
Collection
Publications
Title
Generation of human pulmonary microvascular endothelial cell lines
Journal
Laboratory Investigation
Author(s)
Lehr HA, Krump-Kovalinkova V, Bittinger F, Unger RE, Peters K, Kirkpatrick CJ
ISSN
0023-6837
Publication state
Published
Issued date
2001
Peer-reviewed
Oui
Volume
81
Number
12
Pages
1717-1727
Language
english
Abstract
The limited lifespan of human microvascular endothelial cells in cell culture represents a major obstacle for the study of microvascular pathobiology. To date, no endothelial cell line is available that demonstrates all of the fundamental characteristics of microvascular endothelial cells. We have generated endothelial cell lines from human pulmonary microvascular endothelial cells (HPMEC) isolated from adult donors. HPMEC were cotransfected with a plasmid encoding the catalytic component of telomerase (hTERT) and a plasmid encoding the simian virus 40 (SV40) large T antigen. Cells transfected with either plasmid alone had an extended lifespan, but the cultures eventually entered crisis after several months of proliferation. Only those cells that were transfected with both plasmids acquired the capacity to grow in vitro without demonstrating major crisis, and these cells have been in culture for 24 months. HPMEC isolated from two different donors were used, generating two populations of immortalized cells, HPMEC-ST1 and HPMEC-ST2. Single cell-derived clones of the immortalized cells HPMEC-ST1 exhibited growth characteristics that were similar to those of the parental HPMEC. One selected clone, HPMEC-ST1.6R, displayed all major constitutively expressed and inducible endothelial phenotypic markers, including platelet endothelial cell adhesion molecule (PECAM-1, CD31), von Willebrand factor (vWF), and the adhesion molecules, intercellular adhesion molecule (ICAM-1), vascular adhesion molecule (VCAM-1), and E-selectin. In addition, an angiogenic response was demonstrated by sprout formation on a biological extracellular matrix (Matrigel). The HPMEC-ST1.6R cells did not form tumors in nude mice. The microvascular endothelial cell line, HPMEC-ST1.6R, will be a valuable tool for the study of microvascular endothelial physiology and pathology including gene expression, angiogenesis, and tumorigenesis.
Keywords
, HUMAN EPITHELIAL-CELLS, TELOMERASE ACTIVITY, ADHESION MOLECULE-1, LIFE-SPAN, EXPRESSION, IMMORTALIZATION, CULTURE, LENGTH, SV40, IDENTIFICATION
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Create date
15/12/2011 21:10
Last modification date
20/08/2019 13:55
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