O6 Analysis of angiogenic gene profiling after E-selectin + stem cell therapy in murine ischemic limb

Abstract Introduction There remains a paucity of novel therapeutics for limb salvage in patients with critical ischemia (CLI) whom revascularization procedures have failed and amputation is imminent. We shown that E-selectin+/Mesenchymal Stem Cell (MSC) injections into the ischemic tissue CLI mouse model improves function. Thus, we sought to determine mechanism action E-selectin+/MSC’s pro-angiogenic properties. Methods MSC were extracted from donor mice bone marrow subsequently engineered via viral transduction E-selectin-ires-GFP/AAV GFP/AAV (control) create E-selectin-GFP+/MSC vs GFP+/MSC. Intramuscular E-selectin-GFP+/MSC, GFP+/MSC, or PBS performed hindlimb ischemia. Laser doppler imaging (LDI), confocal laser microscopy, treadmill exhaustion test utilized neovascularization RNA extraction (E-selectin-GFP+/MSC GFP+/MSC) tissues treated GFP+/MSC was performed, followed by RT2 Profiler PCR Array analysis 84 genes involved angiogenesis. served as control. Student’s t-test ANOVA compare means significance set at P < 0.05. Results Compared PBS, treatment increased leg LDI reperfusion (54% vs. 39% 22%, 0.001), distance traversed (162 m 111 110 m, 0.01) footpad vessel density (23% 14% 14%, 0.01). demonstrated gene upregulation occurred 7 (Csf3, Cxcl2, Cxcl5, Serpine1, F2, Lep, Tbx1, Table I.) while tumour necrosis factor (TNF) found be downregulated, when compared tissues. Of these upregulated genes, CXCl2, Leptin T-box1 (Table are likely produced cellular expression profiles also revealed 2-fold more factors Validation functions in-vivo under investigation. Conclusion cell therapy using murine CLI, confers both augmented postnatal The pro-repair effects mediated panel chemokines/cytokines down-regulation TNF E-selectin-GFP+/MSC.