Fitc uea l

A total of 20 ml fresh human umbilical cord blood was obtained from the Obstetrics Department of Shanghai Sixth People's Hospital (Shanghai) and all participants (totally 20 patients; mean age: 24 years old) provided written informed consent. EPCs were isolated from the human umbilical cord blood by Ficoll gradient centrifugation (1,500 × g) for 10 min at room temperature and cultured in endothelial basal medium (Lonza Group Ltd.) containing growth factors (hydrocortisone, 0.2 ml; human basic fibroblast growth factor-B, 2 ml; vascular endothelial growth factor, 0.5 ml; Recombinant human R3 insulin-like growth factor-1, 0.5 ml; human epidermal growth factor, 0.5 ml; ascorbic acid, 0.5 ml; and gentamicin sulfate-amphotericin, 0.5 ml). Isolation, cultivation and identification of EPCs were performed as described previously (6 (link)). Fluorescent staining was used to detect the uptake of Dil-ac-LDL (Molecular Probes; Thermo Fisher Scientific, Inc.) and binding of FITC-UEA-l (Sigma-Aldrich; Merck KGaA). Briefly, the cells were incubated with Dil-ac-LDL (15 µg/ml) for 4 h, and then stained with FITC-UEA-l (10 µg/ml) for 1 h and with DAPI for 5 min at room temperature. The cells were washed three times and analyzed under a fluorescence microscope (Olympus Corporation). EPCs at passages 2–4 were used in subsequent experiments.