Effects of osteopontin on functional activity of late endothelial progenitor cells

Abstract

The aim of this study is to investigate the effect of osteopontin (OPN) on functional activity of late endothelial progenitor cells (EPCs). Total mononuclear cells (MNCs) were isolated from human umbilical cord blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture plates. Late EPCs were positive for both 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine-labeled acetylated low-density lipoprotein (DiI-acLDL) and fluorescein-isothiocyanate-conjugated Ulex europaeus agglutinin lectin (UEA-1). Expression of von Willbrand factor (vWF) and kinase insert domain receptor (KDR) were detected by indirect immunofluorescence staining. Late EPCs of 3-5 passages were treated for 24 h with OPN (to make a series of final concentration: 0.005 µg/ml, 0.01 µg/ml, 0.05 µg/ml, 0.5 µg/ml, 2.5 µg/ml), or vehicle control. The proliferation, migration, and in vitro vasculogenesis activity of late EPCs were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, modified Boyden chamber assay and an in vitro angiogenesis assay, respectively. Late EPCs adhesion assay was performed by replating cells on fibronectin-coated plates, and then adherent cells were counted. Incubation with OPN dose-dependently inhibited the proliferative, adhesive, and in vitro vasculogenesis capacity and increased migratory activity of late EPCs.

PEDF expression regulates the proangiogenic and proinflammatory phenotype of the lung endothelium

“…Osteopontin also has antiadhesive activity. In endothelial progenitor cells, os- teopontin decreases cell adhesion (47). Thus decreased production of tenascin-C and osteopontin may contribute to altered cell adhesion properties of PEDFϪ/Ϫ EC.…”

Expression and localisation of osteopontin and prominin-1 (CD133) in patients with endometriosis

“…This cell subset seems to originate from bone marrow, and to display endothelial progenitor cell-like features (24). The proliferation and migration of endothelial progenitor cells are influenced by OPN (16,17), which may be required for the homing of these cells (25). OPN shows opposite effects depending on tissue and the physiological state of the cell.…”

“…It is known that OPN may influence the angiogenesis, proliferation and migration of endothelial progenitor cells, acting as a regulator of CD133 + progenitor cells (16,17). …”

Lipopolysaccharide Impaired the Functional Activity of Endothelial Colony-Forming Cells

“…It was mainly carried out from December 2010 to July 2012. ECFCs were isolated, cultured and characterized according to previously described techniques [18,19]. Briefly, total MNCs were isolated from umbilical cord blood by Ficoll density gradient centrifugation and cultured on 6-well plates coated with human fibronectin (Chemicon, Billerica, Mass., USA) in endothelial cell basal medium-2 (EBM-2; Lonza, Walkersville, Md., USA) supplemented with EGM-2 MV single aliquots (Lonza) consisting of 5% fetal bovine serum, vascular endothelial growth factor, basic fibroblast growth factor-2, epidermal growth factor, insulin-like growth factor-1, ascorbic acid, gentamicin sulfate, amphotericin-B and hydrocortisone.…”