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PAD is characterized by the occlusion of blood vessels, and its progression results in ischemic ulceration and gangrene, leading to amputation in more than a third of patients. Peripheral arterial disease (PAD) is an indication of systemic atherosclerosis that is undertreated in the United States, and is present in 29% of people over the age of 70 and prevalent in those over the age of 50 with a history of smoking and/or diabetes. Our data suggest that ECFC + MPC delivery could be used to reestablish blood flow in ischemic tissues, and this may be enhanced by coordinated recruitment of host myeloid cells.
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Systemic myeloid cell depletion with anti-Gr-1 antibody blocked the improved blood flow observed with ECFC + MPC and reduced ECFC and MPC retention. Neutrophils declined by day 7, while the number of myeloid cells, macrophages, and monocytes did not. Flow cytometric analysis of ischemic muscles at day 2 revealed increased myeloid lineage cells in ECFC + MPC-injected muscles compared to vehicle-injected muscles. In vivo bioluminescence imaging showed ECFC persisted for 14 days in ECFC + MPC-injected hind limbs. Tail vein injection of human endothelial specific Ulex europaeus agglutinin- I demonstrated an increased number of perfused human vessels in ECFC + MPC compared with ECFC. At day 5, hind limbs injected with ECFC + MPC showed greater blood flow recovery compared with ECFC, MPC, or vehicle. ECFC + MPC, ECFC alone, MPC alone, or vehicle alone were injected into the hind limb ischemic muscle one day after ligation of femoral artery and vein. Here we investigated whether endothelial colony forming cells (ECFC) and mesenchymal progenitor cells (MPC) form vascular networks and restore blood flow in ischemic skeletal muscle, and whether host myeloid cells play a role.