Plasma triacylglycerol-rich lipoproteins (TRL) are being considered as a key lipid fraction in the pathogenesis of atherosclerotic cardiovascular disease. Here we compared the influence of two monounsaturated oils [virgin olive oil (VOO) and high-oleic sunflower oil (HOSO)] on the capability of postprandial TRL to interact with two human vascular cell lines [umbilical vein endothelial (HUVEC) and aorta smooth muscle (HASMC) cells]. A fluorescent probe was used for labeling TRL and to determine receptor activity of HUVEC and HASMC. The values for total cell-associated, bound, and internalized TRL were higher in HUVEC, and TRL from VOO was the better ligand recognized but at lower affinity than TRL from HOSO. There was a competitive effect of very low density lipoproteins (VLDL) for the uptake of TRL by cells, which was found to be dependent on the origin/lipid composition of the ligands and cell-type specific. We also conclude that the VLDL receptor (VLDLr) may contribute significantly to the HASMC binding capacity for postprandial TRL mediated by lipoprotein lipase (LPL) or LPL-binding molecules. Our findings are compatible with a selective role of the clustered O-linked sugar domain of the VLDLr in the catabolism of TRL by human vascular cells.