Apolipoprotein A-I Mimetic Peptide R-D4F Could Improve The Adhesion And Migration Of Endothelial Progenitor Cells Derived From Human Umbilical Cord Blood

Objectives:To establish a method for isolating, culturing and identifying human umbilical cord blood derived EPCs and to investigate whether Apolipoprotein A-I mimetic peptide R-D4F of different concentrations have influences on the function of EPCs in vitro.Methods: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 dishes. After20days cultured, attached cells were isolated and assessed with a laser scanning confocal microscope. Differentiating EPCs were characterized as adherent cells double positive for Dil-ac-LDL and FITC-UEA-I. The influence of R-D4F on EPCs’ adhesion was assayed with adhesion ability experimental determination. EPCs migration was assayed with transwell assay.Results:On the7th day, the adherent cells exhibited the small colonies. After20days cultured, the colonies were expanded, confluenced and displayed a typical "cobblestone" morphology, similar to that of EPCs.On the20th day,90%attached cells took up Dil-ac-LDL, and bound FITC-UEA-I (double positive fluorescence).R-D4F strikingly improved the ability in adhesion and migration of endothelial progenitor cells isolated from human umbilical cord blood (P<0.05). While the concentration of R-D4F was further increased, the improvement effects showed a tendency to enhance. Conclusion:The present study established that R-D4F could improved EPCs’adhesion and migration capability. EPCs derived from human umbilical cord blood may serve as another source of seeding cells for vascular tissue engineering.