Endothelial progenitor cells (EPCs) provide revascularization for many vascular pathologies, including cardiovascular disease, and the expansion of these cells opens up the possibility of their use as a cell therapy. Herein, we reveal that novel combinations of capture and growth molecules strongly expand a population of peripheral blood-derived human endothelial forming cells (ECFCs) to 108 cells from 50 ml peripheral blood within 21 days that exhibit low immunogenicity as well as pro-angiogenic capabilities, making their therapeutic utilization a realistic option. Cell surface marker phenotyping confirmed expression of the progenitor cell markers CD117 and CD34, vascular cell markers VEGFR2, CD144, CD146 and CD31 and absence of leukocyte and mesenchymal markers CD45, CD14, CD11b, CD90 and CD38. Functional experiments revealed that these ECFCs exhibited classical properties of endothelial cells (ECs), namely binding of Ulex europaeus lectin, up-take of acetylated-low density lipoprotein and EC tube formation in vitro. These cells demonstrated a pro-angiogenic phenotype within an independent in vivo rodent model. Taken together, these studies reveal a novel combination of capture and expansion molecules for human peripheral blood EPC expansion with clear pro-angiogenic properties, in vitro and in vivo and thus may provide clinical utility for humans in the future.
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