Phenotypic Characterization of Human Umbilical Vein Endothelial (Ecv304) and Urinary Carcinoma (T24) Cells: Endothelial Versus Epithelial Features

Abstract

ECV304 cells reported as originating from human umbilical vein endothelial cells by spontaneous transformation have been used as a model cell line for endothelia over the last decade. Recently, deoxyribonucleic acid fingerprinting revealed an identical genotype for ECV304 and T24 cells (urinary bladder carcinoma cell line). In order to resolve the apparent discrepancy between the identical genotype and the fact that ECV304 cells phenotypically show important endothelial characteristics, a comparative study was performed. Immortalized porcine brain microvascular endothelial cells/C1-2, and Madin Darby canine kidney cells were included as typical endothelial and epithelial cells, respectively. Various methods, such as confocal laser scanning microscopy. Western blot, and protein activity tests, were used to study the cell lines. ECV304 and T24 cells differ in criteria, such as growth behavior, cytoarchitecture, tight junction arrangement. transmembrane electrical resistance, and activity of gamma-glutamyltransferase. Several endothelial markers (von Willebrand factor, uptake of low-density lipoprotein, vimentin) could clearly be identified in ECV304, but not in T24 cells. Desmoglein and cytokeratin, both known as epithelial markers, were found in ECV304 as well as in T24 tells. However, differences were found for the two cell lines with respect to the type of cytokeratin: in ECV304 cells mainly cytokeratin 18 (45 kDa) is found, whereas in T24 cells cytokeratin 8 (52 kDa) is predominant. As we could demonstrate, the ECV304 cell line exposes many endothelial features which, in view of the scarcity of suitable endothelial cell lines, still make it an attractive in vitro model for endothelia.

Vaccination with xenogeneic tumor endothelial proteins isolated in situ inhibits tumor angiogenesis and spontaneous metastasis

“…45,46 Nevertheless, ECV304 shows many endothelial characteristics, resembles Cl-2, a porcine brain microvascular endothelial cells and differs from T24 in phenotype. 47 ECV304 cells not only show endothelial morphology, but also express endothelial markers, uptake Ac-LDL and undergo tube formation in Matrigel. 47,48 So ECV304 is still frequently used as an endothelial-like cell line.…”

“…47 ECV304 cells not only show endothelial morphology, but also express endothelial markers, uptake Ac-LDL and undergo tube formation in Matrigel. 47,48 So ECV304 is still frequently used as an endothelial-like cell line. In this study, we used ECV304 rather than HUVEC in the cell migration assay, because HUVECs are extremely sensitive to serum-free condition and die quickly without the presence of serum, but the depletion of serum in the medium during the assay is necessary to avoid the interference of cell growth and division.…”

L-methionine reduces oxidant stress in endothelial cells: Role of heme oxygenase-1, ferritin, and nitric oxide

“…20 ECV304 endothelial cells were maintained and subcultured in M199 medium containing 10% fetal bovine serum and gentamycin (50 µg/mL). The cells were grown in a humidified incubator at 37°C and 5% CO 2 .…”

Protein tyrosine phosphatase hPTPN20a is targeted to sites of actin polymerization

“…African-green-monkey COS1 and HEK (human embryonic kidney)-293 cells, as well as human HeLa cervical adenocarcinoma, U2OS osteosarcoma, U87MG glioblastoma, MDA-MB-231 breast adenocarcinoma and ECV304 urinary carcinoma [21,22] cells were cultured at 37…”