Background and Objective; The adipose-derived stem cells (ADSCs) for repairing damaged tissue has become a potential new therapy in various diseases related to cell loss. We tested the effect of genetically engineered ADSCs to secrete EPO in a rat ischemic myocardium compared to wild type ADSCs.
Methods; Recombinant Lentiviral vectors containing the GFP (Lenti-GFP) or rat EPO gene (Lenti-EPO) were made by co-transfection of 293T cell. We identified GFP expression in rat ADSCs infected Lenti-GFP (rADSC/Lenti-GFP) and measured EPO level at the supernatant of rat ADSC infected Lenti-EPO (rADSC/Lenti-EPO). In vitro, tube formation was evaluated by Matrigel assay. FACS analysis was evaluated by dual chamber system with CMC and rADSC under the hypoxia injury. EPO receptor (EPO-R) expression and survival signals were evaluated by Western blot. Ischemia - reperfusion injury was performed to heart with reperfusion follow by 4 hour ligation of coronary artery in male SD rats. DAPI-labeled rADSC/Lenti-GFP (1x10⁶ cells, n=5) and rADSC/Lenti-EPO (1x10⁶ cells, n=5) were injected in vicinity of infarction area. GFP and DAPI expression, hemodynamic factor, Western blot and angiogenesis were evaluated on 2 weeks after cell injection.
Results; Higher EPO level in media and increased tube formation was observed in rADSC/Lenti-EPO group than that of rADSC/Lenti-GFP group. Apoptosis under the hypoxia decreased and EPO-R, JAK 2, STAT 3 and p-Akt expression increased in both CMC and rADSC infected Lenti-EPO. Heart function and angiogenesis was increased in hADSC/Lenti-EPO group. Expressions of EPO-R, JAK 2, STAT 3 and p-Akt were enhanced at hADSC/Lenti-EPO group (Fig).
Conclusion; These results suggest that ADSCs combined with EPO gene transfer may be a useful strategy for ischemic heart disease.