Assessment of ability of human adipose derived stem cells for long term overexpression of IFN-β and LIF as therapeutic cytokines

Introduction: Adipose-derived stem cells (ADSCs) are a subset of MSCs that their therapeutic effects in various diseases make them an interesting tool in cell therapy. ADSCs can be genetically engineered by lentiviral vectors for cell-based therapy purposes.Objectives: In the current study we aimed to overexpress IFN-β and LIF cytokines in human ADSCs and assess the ability of transduced ADSCs for long-term expression of IFN-β and LIF, simultaneously.Materials & Methods: Here, we designed a construct containing IFN-β and LIF and then, transduced characterized hADSCs by this construct via a lentiviral vector (PCDH-513B). We assessed the ability of long term expression of the transgene in transduced cells by western blotting and ELISA techniques on days 15, 45 and 75 after transduction.Results: According to the obtained data, 95.8% and 94.7% of isolated hADSCs cells were positive for expression of CD73 and CD105, respectively while the expression of CD45, as a negative marker, was only 5.85% and isolated hADSCs successfully differentiated into osteocytes and adipocytes. In addition, our results showed high-efficiency transduction with highest expression rates on day 75 after transduction which were 70 pg/ml for IFN-β and 77.9 pg/ml for LIF compared with 25.60 pg/ml and 27.63 pg/ml, respectively in un-transducted cells (p=0.0001).Conclusion: All in all, we successfully constructed a hADSC population stably overexpress IFN-β and LIF cytokines. Considering the IFN-β and LIF anti-inflammatory and neuroprotective effects as well as immune-regulatory properties of hADSCs, the obtained cells of this study could be subjected for further detailed evaluations of their impact on the EAE mice model.

کلیدواژه ها:

Mazdak Ganjalikhani-Hakemi ، PhD ، Associate Professor ، Department of Immunology ، School of Medicine ، Isfahan University of Medical Sciences ، Isfahan ، Iran