Ab201336

Human Umbilical Endothelial Cells (HUVECs, Gibco) were cultured in Medium 200 (MÀ200, Gibco) supplemented with Large Vessel Endothelial Supplement (LVES), at 37 °C with 5 % of CO 2 . Upon confluence of 80 %, HUVECs were trypsinized and cultured on the coated substrates at a density of 5000 cells/cm 2 (for evaluation of cell morphology and adhesion) or 50,000 cells/cm 2 (for evaluation of reendothelization) using Low Serum Growth Supplement (LSGS, Gibco). After 24 h of culture, cell viability was quantified with Alamar Blue (BIO-RAD) following the manufacturer's instructions. The substrates were then washed with PBS and fixed with 10 % formalin for the following immunocytochemical analysis. The expression of von Willebrand factor (vWF), a key endothe-lial marker, was visualized by staining with anti-vWF antibody (0.5 mg/mL, ab201336, abcam) and secondary antibody antimouse IgG1 (Alexa Fluor 488, 1:400, ThermoFisher Scientific). Nuclei were stained with DAPI (1:500) and actin filaments with phalloidin (1:50). Images were taken with a confocal laser scanning microscope (TCS SP8, Leica). Cell morphology analysis was performed with Fiji (Image J). TCPS was used as a control substrate.