Characterization of ovine umbilical vein endothelial cells and their expression of cell adhesion molecules:...

This paper reports the isolation and characterization of sheep umbilical vein endothelial cells (ShUVEC) and examines the kinetics of the expression of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). In culture, the cells demonstrate the classical endothelial cell phenotype, including the expression of von Willebrand Factor (vWF), the metabolism of acetylated-low density lipoprotein (Ac-LDL) and the typical cobblestone monolayer appearance. Northern analysis of the expression of ICAM-1 and VCAM-1 mRNA by ShUVEC stimulated with either LPS or recombinant ovine (ro)-TNF-alpha or IL-1 beta demonstrated peak RNA expression for both molecules at between 2 and 6 h after stimulation. Maximum VCAM-1 protein expression occurred between 6 and 12 h after stimulation with ro-TNF-alpha, followed by a slight decrease to a plateau extending beyond 48 h. The levels and kinetics of expression of additional cell surface proteins on ShUVEC, including P-selectin (CD62P), beta 1 integrin (CD29) and ovine MHC classes I and II, were found to be almost identical to that observed on human endothelial cells (HUVEC) cultured under similar conditions. Based on the above results, we conclude that the ShUVEC represent the ovine equivalent of HUVEC, both phenotypically and functionally.