Study the Effect of Differentiated Endothelial Progenitor Cells on Experimental Model of Peripheral Ischemia

Background: The adult vasculature possesses the ability to remodel itself in response to a variety of stimuli, including hemodynamic forces (shear) and tissue metabolic demands (ischemia).Objectives: We aimed to study the effect of human cord blood Endothelial Progenitor Cells (hEPCs) on angiogenesis in a rat model of unilateral peripheral vascular disease (unilateral femoral artery ligation).Methods: The work divided into 2 parts in vitro one for cells isolation, characterization and differentiation followed by second in vivo part for cells transplantation in surgical induced peripheral ischemia. Animals were grouped; control sham-operated rats (n=12), ischemia group (n=12) subjected to surgical left femoral artery ligation with no further intervention, and a third ischemic group (n=12) subjected to surgical left femoral artery ligation followed by 24hr transplantation with differentiated hEPCs (IM injection of 5 X 106). In vitro, hEPCs were isolated from human cord blood. The cells characterization was assessed by flow cytometry. After rat scarification, muscle tissues of all studied groups were harvested for pathological assessment and for Vascular Endothelial Growth Factor (VEGFR2), von Willbirand Factor (vWF) genes expression using quantitative real time PCR.Results: In vitro part of the study showed successful isolation of hEPCs. The hEPCs were identified by having CD133+, CD34+ and KDR. Ischemic group transplanted with differentiated hEPCs group showed an improvement in blood vessels formation, detected by high significant level of VEGFR2, and vWF genes expression and histopathological angiogenesis compared to ischemic group without further transplantation.Conclusion: EPCs can be isolated and identified from human cord blood. Differentiated EPCs showed successful transplantation and blood vessel formation in ischemic experimental animals.