Infection of MSCs by adenovirus expression system expressing NRF2 as a cytoprotective factor

Poor viability of Mesenchymal Stem Cells [MSCs] following transplantation is one of the major challenges in their therapeutic application. Manipulation of MSCs by the genetic engineering method is one of the strategies used to protect the cells against cytotoxic microenvironment. However, maintaining multi differentiation capacity of MSCs following manipulation is important. We investigated if the manipulation of MSCs with NRF2 affects the multi differentiation capacity. MSCs were isolated from bone marrow. NRF2 was isolated and TOPO cloned into the pENTR vector. The recombinant vector was transferred into pAD/CMV/V5-DEST vector by gateway technology. Recombinant adenovirus was produced in AD293 cells, followed by being infected into MSCs. Expression of NRF2 was verified by RT-PCR. The NRF2 engineered MSCs were exposed to stress conditions followed by the evaluation of the cells viability and apoptosis. Finally, NRF2 expressing MSCs differentiation into osteoblast and adipocyte lineages was studied. NRF2 was successfully expressed in MSCs. NRF2- MSCs differentiation into osteoblast and adipocyte lineages indicating overexpression of NRF2 does not affect the differentiation property of MSCs. Expression of NRF2, a well known cytoprotective factor, by using adenovirus expression system does not intervene in the differentiation capacity of MSCs. NRF2-MSCs might be applicable for stem cell-based cell therapy in future

[Application of expired platelets in the preparation of platelet gel and study of proliferative effects of expired platelet derived growth factor on variety of cell lines]

There is growing evidence indicating that growth factors derived from platelets can be used in wound healing. This study aimed to investigate whether old platelets can be used as the main material for preparation of platelet gel and as substitute for FBS and FCS in cell culture medium. In this exprimental study, platelets were prepared from voluntary blood donors by centrifugation. To prove the hypothesis that the platelet gel and the growth factor derived from expired platelets are able to propagate different cells, platelet derived factors were prepared from both new and expired platelet-rich plasma. The concentration of platelet-derived growth factors was measured by ELISA and cell proliferation was measured by MTT assay. The results showed the high quality of platelet gel obtained from old platelets. Our results also revealed that old platelets released growth factors similar to those released by new platelets. The growth factors derived from old and new platelets had the same proliferation effects on MSC, CHO, and Fibroblast cell lines Old platelets released the same growth factors that new platelets did; this showed that old platelets as valuable constituents of blood are cost effective to be used