Experimental study on rabbit endothelial progenitor cells-seeded stents in vitro / 中国介入心脏病学杂志

ABSTRACT

Objective To fabricate rabbit endothelial progenitor cells(EPCs)-seeded stents in vitro and evaluate its feasibility.Methods The mononuclear cells of rabbit were separated by density-gradient centrifugation using lymphocyte isolation.Cells were seeded on fibronectin-coated 6-well plates and maintained on M199 with endothelial cell growth supplements(ECGS,30 ?g/mL).After 2 weeks,cultured cells were identified by immunofluorescence and immunocytochemistry.EPCs proliferation and migration were measured by drawing a growth curve and modified Boyden chamber assay.EPCs adhesion assay was performed by replating EPCs on fibronectin-coated dishes.VEGF and G-CSF were assayed by ELISA while NO was tested by nitrate reductase method.EPCs were poured on fibronectin-coated or uncoated stents.After 6 days,scanning electron microscopy and fluorescence microscopy observation were performed.Results About 14 days after seeding,a majority of spindle-like cells appeared which were characterized as cells,double positive for Dil-acLDL and FITC-UEA-I.EPCs were further confirmed by its expression of the endothelial cell marks of vWF.After 2 weeks culture,EPCs proliferated in exponential growth.They showed high ability of adhesion(96.7?3.2%),migration(15?4) and could secret VEGF,G-CSF and NO.Compared with uncoated stents group,more EPCs migrated and adhered on fibronectin-coated stents group(27.80?4.26 vs 6.10?3.07cells/?400 field,P