Effects of homocysteine on number and activity of endothelial progenitor cells from peripheral blood

• Published in: Journal of molecular and cellular cardiology Vol. 36; no. 2; pp. 233 - 239
• Main Authors: Chen, J.Z., Zhu, J.H., Wang, X.X., Xie, X.D., Sun, J., Shang, Y.P., Guo, X.G., Dai, H.M., Hu, S.J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.02.2004
• Subjects: Cell Division - drug effects; Cell Movement - drug effects; Cell Movement - physiology; Coronary artery disease; Endothelial dysfunction; Endothelial progenitor cell; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Endothelium, Vascular - physiology; Fluorescent Dyes; Homocysteine; Homocysteine - pharmacology; Humans; Laser scanning confocal microscope; Monocytes - cytology; Monocytes - drug effects; Monocytes - physiology; Stem Cells - cytology; Stem Cells - drug effects; Stem Cells - physiology; Endothelial dysfunction; Endothelial progenitor cell; Homocysteine; Coronary artery disease; Laser scanning confocal microscope
• ISSN: 0022-2828; 1095-8584
• DOI: 10.1016/j.yjmcc.2003.10.005

The aim of this study is to investigate whether homocysteine (Hcy) has influences on endothelial progenitor cells (EPCs) number and activity. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 d cultured, attached cells were stimulated with Hcy (to make a series of final concentrations: 10, 50, 100 and 200 μmol/l) or vehicle control for the respective time points (6, 12, 24 and 48 h). EPCs were characterized as adherent cells double positive for DiLDL uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, and then adherent cells were counted. Incubation of isolated human MNCs with Hcy dose and time dependently decreased the number of EPCs, maximum at 200 μmol/l, 24 h (approximately 50% reduction, P < 0.01). In addition, Hcy dose and time dependently impaired EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity. In conclusion, hyperHcy may induce the reduction of EPCs with decreased functional activity.