A Study Of Transplanting Endothelial Progenitor Cells To Protect The Ischemia-Reperfusion Injury Of Rat After Liver Graft

Objective: Successfully isolate the bone marrow mononuclear cellsfor culture and induce it to EPCs, immunocytochemical stain and doublefluorescence stain to identify; Successfully use PKH26GL to stain the EPCsfor stem cell transplantation; Successfully establish the rat OLT model tostudy the liver IRI and observe whether the EPCs can enter into the rathepatic sinusoid after the EPC transplanting; Observe the affecting of liverand liver function after transplantation.Methods: Used percoll density gradient centrifugation to isolate thebone marrow mononuclear cells from the femoral and tibial bone marrow,then cultured in selective DMEM medium to induce； Observed themorphological changes and the expression of the cell markers byimmunocytochemistry, stained with Dil-ac-LDL and FITC-UEA-1, andmarked by the red fluorescent dye PKH26GL for the following experiment;48female SD rats were randomly divided into two groups and used themodified Kamada cuff technique to establish the rat OLT models; One group was OLT+EPCs transplantation group and the other was control group onlywith OLT, then the EPC group rats were transported with EPCs via the tailvein and the control group were transported with saline; Both groups takedthe blood samples and the liver tissue samples after6hours,24hours,48hours to detect the ALT level, the serum TNF-α level, the EPC homing，theEPC antigen expression, and the liver tissue injury.Results: Successful isolated the rat bone marrow mononuclear cellsand inducted by1week; morphologically observing, immune cells stain, anddouble stain were identified as the EPCs; Compared with the control group,fluorescence microscope could detect the homing of EPCs and the EPCantigen could be positive express in the liver tissue at all time point；Compared with the control group, the liver injury was lighter, the liverfunction improved significantly and the expression of inflammatorycytokines in the serum was less in the EPC group(p <0.05).Conclusion: Percoll density gradient centrifugation method anddifferentiation culture can get high quantity and purity EPCs; Aftertransplantation, the EPCs could homing in the liver tissue to repair thedamage of the liver sinusoids, thus improve the microcirculation, andpartially release the inflammation, so ultimately improve the ischemia-reperfusion injury after liver graft.