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VARIABILITY IN E‐SELECTIN EXPRESSION, mRNA LEVELS AND sE‐SELECTIN RELEASE BETWEEN ENDOTHELIAL CELL LINES AND PRIMARY ENDOTHELIAL CELLS
Author(s) -
Galley H.F,
Blaylock M.G,
Dubbels A.M,
Webster N.R
Publication year - 2000
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1006/cbir.1999.0455
Subject(s) - umbilical vein , e selectin , flow cytometry , cell culture , human umbilical vein endothelial cell , endothelium , tumor necrosis factor alpha , endothelial stem cell , microbiology and biotechnology , immunofluorescence , cell , biology , cell adhesion , immunology , chemistry , in vitro , endocrinology , biochemistry , antibody , genetics
Endothelial cell lines express markers and are assumed to exhibit other endothelial cell responses. We investigated E‐selectin expression from human umbilical vein endothelial cells, the spontaneously transformed ECV304 line and the hybrid line EA.hy926 by flow cytometry and immunofluorescence, mRNA and soluble E‐selectin release. In cells exposed to tumour necrosis factor α (TNF‐α) and interleukin‐1β (IL‐1β), median (range) percentage of E‐selectin‐positive HUVECs increased from 1.6(0.9–6.2)% to 91.4(83.0–96.1)%, ( P =0.001) using flow cytometry. In contrast, E‐selectin expression by ECV304 and EA.hy926 cell lines was 100‐fold lower. E‐selectin mRNA was detectable after 2h, maximal at 6h in HUVECs and undetectable in EA.hy926 and ECV304 cell lines after exposure to TNF‐α/IL‐1β. sE‐selectin accumulation increased ( P =0.004) in HUVECs only. Neutrophil adherence to ECV304 and EA.hy926 cells was poor compared to HUVECs ( P =0.004). The cell lines ECV304 and EA.hy926 do not exhibit normal endothelium expression of E‐selectin, and may not be appropriate for studies of adhesion.