The Effects of Hif-1 Gene on the Expression of IL-10, TGF-β and Versican Genes in Human Bone Marrow and Adipose Tissue Mesenchymal Stem

Mesenchymal stem cells (MSCs) derived from bone marrow and adipose tissues are often hypoxic. These cells are important in tissue repair as well as in the secretion of anti-inflammatory factors and reduction of inflammation. IL-10 and TGF-β are anti-inflammatory cytokines and play an important role in the immune system by modulating inflammation. Versican is a protein that plays an important role in repairing damaged tissues. In this study, the effect of hypoxia on expression of IL-10, TGF-β and versican genes in MSCs was studied in comparison with normoxia. Flow cytometry for surface markers (CD105, CD73 and CD90) was used to confirm MSCs. Desferrioxamine and cobalt chloride (200 µM and 300 µM final concentrations respectively) were added to the cell culture media to induce hypoxia-mimetic conditions. Western Blotting was used to detect HIF-1α protein and confirm the hypoxia-mimetic conditions in MSCs. The expression of VEGF, IL-10, TGF-β and versican genes in MSCs was evaluated using RT-PCR. Microscopic examination and results of flow cytometry confirmed the mesenchymal nature of the cells. Western blotting results also showed that the expression of HIF-1α gene in MSCs increased significantly under hypoxia-mimetic conditions. RT-PCR analyses showed that the expression of VEGF, IL-10, TGF-β and versican genes in hypoxic conditions increased significantly compared to normoxia in both mesenchymal cells derived from bone marrow and from adipose tissues. The results of this study showed that MSCs in hypoxia can repair tissue and inhibit inflammation by expressing anti-inflammatory cytokines and versican restorative factor.