The effect of differentiation inducing factors on osteoblastogenesis from mesenchymal stem cells

Mesenchymal Stem Cells are pluripotent cells differentiating through development from mesenchymal cells.This cells have the potential to differentiate to osteoblasts, osteocytes, adipocytes and cartilage cells. In vitro differentiation to osteoblasts needs various factors to produce a microenvironment mimicking the natural microenvironment in bone tissue. In this research the effect of Dexamethazone, ascorbic acide and beta glycerolphosphate on the differentiation process of bone marrow mesenchymal stem cells to osteocytes is investigated. Mesenchymal stem cells were separated from 2 ml aspirated bone marrow using ficole, HBSS, and the attachment ability of mesenchymal stem cells to plastic surfaces. Different markers and Alizarin red staining were implemented to study the success of in vitro differentiation Mesenchymal stem cells were fusiform in the beginning and after differentiation the phenotype changed to calcium producing cells. Staining with Alizarin red confirmed this process. RT-PCR on osteocalcin and alkalin phosphatase showed a change from negative expression to positive expression. These results show that in vitro osteoblastogenesis from mesenchymal stem cells is confirmed and these cells could be implemented as a model to study genetic and epigenetic factors in the differentiation cells.