Background: Recent studies have demonstrated that adipose-derived mesenchymal stem cells (MSCs) can attenuate ischemic tissue injury, possibly by their paracrine effect or differentiation into local cell types. But homing of transplanted cells either transfused systematically or via direct intra-lesional injection, is reported to be very low and transient, which reduces the therapeutic effects. CXCR4 is identified as crucial for homing and engraftment of stem cells. In this study, we overexpressed CXCR4 on Adipose-derive MSC to enhance their level of engraftment and perhaps improve their translational effectiveness. Materials and Methods: Wild type variant of CXCR4 gene was chemically synthesized and cloned in pCDH-513b lentiviral vector. Viral particles were generated and their concentrations were determined according to Trono lab protocol after transfection of HEK293 cells. Adipose mesenchymal stem cells in passage 3 and 4 were transduced with CXCR4. 72 h Post transduction, cells were selected with puromycin for 3 days. Migration ability of transduced cells was assessed by transwell chamber assay. Results: Molecular cloning of synthetic genes fragments was confirmed by DNA sequencing. The presence of GFP positive MSCs confirmed the high efficiency of transduction. Our qPCR result showed overexpression of CXCR4 at mRNA level in transduced MSCs. Genetically modified MSCs significantly gain enhanced migration capabilities in comparison with control group (p<0.05). Conclusion: This strategy represents a step to improve MSCs‟ homing and overcome one of the hurdles of cell therapy.

Keywords