Cell adhesion markers are expressed by a stable human endothelial cell line transformed by the SV40 large T antigen under vimentin promoter control

Abstract

Markers of endothelium have been studied in a new endothelial cell line derived from human umbilical cord vein cells by microinjection of a recombinant gene that includes a deletion mutant of the human vimentin gene regulatory region controlling the large T and small t antigen coding region of the SV40 virus. In culture, this immortalized venous endothelial cell line (IVEC) demonstrated morphological characteristics of endothelium; uptake of acetylated low density lipoprotein and presence of the Factor VIII-related antigen. Treatment of IVEC cells with Interleukin-1 beta (IL-1 beta) at 10 U.ml-1 activates the expression of cell adhesion molecules such as endothelial leucocyte adhesion molecule (ELAM-1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1), as observed in primary culture. Prostacyclin secretion was induced in the IVEC cells by 100 nM PMA treatment and thrombin at 0.5 U/ml. Angiotensin converting enzyme (ACE) activity detected in IVEC cells was present but lower than ACE activity in primary endothelial cells and was completely blocked by enalaprilat (1 microM), a specific ACE inhibitor. The presence of ACE mRNA was also demonstrated in IVEC cells by RT-PCR amplification. Our data demonstrate that endothelial cells immortalized by use of this recombinant gene retain the morphological organization and numerous differentiated properties of endothelium.

Immortalized bone‐marrow derived pig endothelial cells

“…Although BM‐derived EC may be crucial for the engraftment of porcine BM during the induction of mixed chimerism [1], to date there is no report on the generation of a porcine EC line derived from the BM microvasculature. In contrast, such lines have been established from murine and human BM [34–36]. In this view, it was of relevance to generate immortalized EC lines from both large vessels and the microvasculature with preserved EC properties and a well‐defined phenotype including the MHC haplotype.…”

Plasmodium falciparum domain mediating adhesion to chondroitin sulfate A: A receptor for human placental infection

“…A stable transfectant of Chinese hamster ovary (CHO)-745 cells (CSA negative) (12) permanently expressing cDNA of CD36 (13), ICAM-1 (14), vascular cell adhesion molecule 1 (VCAM-1) (15), and E-selectin (16) was constructed by using Fugene 6 transfection reagent (Roche Diagnostics). A stably transformed human umbilical vein endothelial cell line was kindly provided by D. Paulin and P. Vicart (Institut Pasteur) (17). Surface expression of platelet endothelial cell adhesion molecule 1, ICAM-1, E-selectin, and VCAM-1 was analyzed by using specific mAbs (R & D Systems).…”

Characterization of Novel Clonal Murine Endothelial Cell Lines With an Extended Life Span

“…To circumvent the limitations associated with primary EC cultures, many laboratories have established immortalized EC lines. Transfection or infection with viral oncogenes has allowed generation of EC lines endowed with an extended life span, but which have frequently lost endothelial-specific differentiation properties and thus differ considerably from their normal counterparts (Williams et al, 1988;O'Connell and Edidin, 1990;Durieu-Trautmann et al, 1991;Harder et al, 1991;Ades et al, 1992;Fickling et al, 1992;Vicart et al, 1993;Roux et al, 1994;Fontijn et al, 1995; 1 To whom correspondence should be addressed at E-mail: Roberto.Montesano@medecine.unige.ch Lechardeur et al, 1995;Candal et al, 1996;Kanda et al, 1996;Moldovan et al, 1996;Schweitzer et al, 1997). On the other hand, only a few spontaneously immortalized EC lines are available (Gumkowski et al, 1987;Cockerill et al, 1994;Bastaki et al, 1997).…”