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late EPCs, which emerged 2�C4 weeks after the start ofMNC culture. The characterization of EPCs was conducted as previously described. The tube formation assay was performed on EPCs to assess their angiogenic capacity, which is involved in new vessel formation. The in vitro tube formation assay was performed using the Angiogenesis Assay Kit according to the manufacturer��s protocol. In brief, the ECMatrix gel solution was thawed at 4 overnight, mixed with the ECMatrix diluent buffer, and placed in a 96-well plate at 37 for 1 hour to allow the matrix solution to solidify. EPCs were treated with or without 2.5�C10 ��Mof atorvastatin or rosuvastatin for 24 hours and then harvested. A total of 104 cells were placed on the matrix solution with 10 ng/mL recombinant human stromal cell-derived factor 1 , and the samples were incubated at 37 for 12 hours. The na?ve EPCs group was used as the negative control. Tube formation was inspected under an inverted light microscope. Images of four representative Sirtuin modulator 1 fields were taken, and the averages of the branching points formed by the cells were quantified. The migratory function of late EPCs, which is essential for vasculogenesis, was evaluated by a modified Boyden transwell chamber assay. Late EPCs were incubated with 10 ��Matorvastatin or rosuvastatin for 24 h. Then, 4×104 treated EPCs were placed in the upper chamber of 24-well Transwell plates with polycarbonate membranes in EGM-2 MV medium. EGM-2MV medium with VEGF and SDF-1 was placed in the lower chamber. After incubation for 24 h, the membranes were washed briefly with PBS and fixed with 4 paraformaldehyde. The upper sides of the membranes were wiped gently with a cotton ball. The membranes were then stained using hematoxylin solution and removed. The magnitude of late EPC migration was evaluated by counting the migrated cells in six random high-power microscope fields. To evaluate the angiogenic effects of atorvastatin and rosuvastatin, we performing unilateral hindlimb ischemia surgery on wild-type male ICR mice. Ligation and 1624602-30-7 customer reviews excision of the femoral artery effectively blocked blood flow in ICR mice. Blood flow in ischemic limbs was significantly recovered in the

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