Vascular Endothelial Progenitor Cells Transferred By HGF Gene Promote Angiogenesis In Vivo

ObjectiveMononuclear cells were isolated from rats bone marrow, then special Medium was used to make them differentiate to endothelial progenitor cells(EPCs) hepatocyte growth factor (HGF) gene were transfered into endothelial progenitor cells (EPCs) of rats primary cultured in vitro; Then the EPCs transferred by HGF gene were transplanted to null rats with ischemia hindlimb to detect the angiogenesis in vivo.Methods1. Bone marorw mononuclear cells(BMMNCs) were obtained from bone marrow of4-6w Wistar rats (120-150g) by flushing rat’s femur and tibia and were isolated by density gradient centrifugation.cultured in10%FBS DMEM with recombined human VEGF, FGF.2. Endothelial progenitor cells were transferred by HGF gene with defective adenovirus (Ad-GFP) as a carrier and transfection efficiency were identified.3.The null rats ischemia hindlimb model were established, and were aparted to3group:Ad-HGF-EPCs group which were transplanted by EPCs transferred by HGF gene, EPCs group which were transplanted by EPCs. control group that were injected to equal NS. The expression of HGF protein was detected by ELISA. The blood perfusion of ischemia hindlimb were detected by LDPI. and the MVP were detected by CD31immunohistochemistry.Results1. Rat EPCs were isolated and cultured successfully in vitro.2. HGF were transfected into primary cultured EPCs successfully, green fluorescent protein(GFP) were observed through fluorescence microscope. The efficient of transfection was above80%72h later.3. The null rats ischemia hindlimb model were established successfully. The function scores of ischemia hindlimb in three groups raised quickly and achieved tip in2days after operation. The rate of limb autotomy and necrosis reduced after transplanted by EPCs tranfered by HGF gene. The function score of ischemia hinlimb was less in Ad-HGF-EPCs group than EPCs group. HGF protein were over expressed in plasm of Ad-HGF-EPCs group. The ischemia hindlimb of Ad-HGF-EPCs group had more blood perfusion than EPCs group, and vascular density was higher.ConclusionsThe ability of angiogenesis of EPCs transferred by HGF gene were greater than EPCs in vivo.