Hypoxia-inducible factor over-expression on human peripheral blood endothelial progenitor cells in vitro.docVIP

 PAGE \* MERGEFORMAT 2 Hypoxia-inducible factor over-expression on human peripheral blood endothelial progenitor cells in vitro Authors: Meng Jiang, Wang Qian, Bin Yao, Ben He, Qin Shao, Wong Ting-9 [Abstract] In order to study hypoxia-inducible factor (hypoxia inducible factor-1alpha, HIF-1α ) in vitro on human peripheral blood progenitor cells (endothelial progenitor cells, EPC) to vascular endothelial cells (endothelial cells, EC) differentiation, by density gradient centrifugation from human peripheral blood EPC, Electroporation HIF-1α plasmid transfected to the EPC, calculate transfection efficiency, RT-PCR was determined HIF-1α , HIF-1β and HIF-1α downstream target gene vascular endothelial growth factor (vascular endothelial growth factor, VEGF) mRNA in the plasmid content changes before and after transfection, immunocytochemistry to detect the under normoxic HIF-1α protein in HIF-1α plasmid transfected protein expression before and after different time points, cell surface antigen clusters detected by flow cytometry FITC-CD31 EPCs / EC in the non-transfected group, pEGFP empty vector or the HIF-1α plasmid transfected group transfected 3-10 days after the difference between the groups observed under light microscope before and after transfection of cell morphology and the degree of differentiation. The results showed that: EPC obtained after transfected by electroporation, plasmid transfection efficiency of about 20%; RT-PCR showed HIF-1α mRNA expression in normoxia there, and over-expression of HIF-1α content increased (P