Isolation and Characterization of EPCs

EPCs were obtained from mononuclear cells (MNCs) as the previous reports [23 (link)]. MNCs were discontinued into -complement medium (Gibco, USA) with endothelial growth factors (Sigma-Aldrich, USA), 10% FBS (Gibco, USA) and seeded on fibronectin (50 μg/mL, Sigma-Aldrich, USA)-pre-coated 6-well dishes (Sigma-Aldrich, USA). Cells were cultured at the condition of 5% CO₂ and 37° C. Dead cells were removed after three days, and then the culture mediums were refreshed every three days. And the EPCs were identified and characterized by DiI-acLDL and bind fluorescein iso-thiocyanate UEA-1 using immunofluorescent analysis after two weeks. The SAG, cyclopamine, and Y294002 were purchased (Sigma-Aldrich, USA).