Elevated expression of stemness genes in adipose-derived mesenchymal stem cells cultured on fibrin scaffold

Abstract

In regenerative medicine, MSCs need to be pluripotent for better results. In this study, the effect of fibrin scaffold on expression of stemness genes was examined. Adipose-derived MSCs were cultured in tissue culture plates (2D) and 3-dimensional (3D) fibrin scaffolds. The effect of fibrin scaffold on proliferation of adiposederived MSCs was evaluated by MTT assay. The expression of stemness genes (OCT4 and SOX2) were evaluated by qRT-PCR, and flow cytometry was done for Nanog protein level. Cultured MSCs on fibrin scaffold were able to proliferate according to data obtained by MTT assay. Expression of OCT4 and SOX2 had a significant increase in cells were cultured in 3D condition compared to 2D condition (P < 0.05). Also, increased expression of Nanog protein in 3D culture was observed (P < 0.05). OCT4 and SOX2 in 3D condition increased two-fold and three-fold respectively in 2D and 3D conditions. Moreover, expression of Nanog increased 30% more than in 2D condition. Evaluation of important pluripotency regulators such as OCT4, SOX2, and Nanog showed that fibrin scaffolds are useful instruments to maintain stemness of MSCs, which is essential in field of stem cell therapy and regenerative medicine.