Isolation and differentiation of murine mesenchymal stem cells into osteoblasts in the presence and absence of dexamethasone

Bone marrow is rich source of multipotent stem cells that can be easily isolated and differentiated. mesenchymal stem cells (mscs) from the bone marrow have multilineage potential and have many therapeutic applications in the form of cellular therapy,tissue repair and regenerative medicine. mscs differentiation into osteoblasts is a complex process that is regulated by many internal and external factors. we studied the differentiation of mscs isolated from tibia and femur of mouse into osteoblasts using osteoblast inductive medium in the presence and absence of dexamethasone. alkaline phosphatase (alp) activity,total protein concentration,total cell number and mineralization was checked during differentiation process in the presence and absence of dexamethasone. total protein of induced and un-induced cells were also analyzed. alp activity was higher in induced cells as compared to un-induced cells but total protein concentration and cell number was higher in un-induced cells. after differentiation the induced cells were positive for alizarin red stain even in the absence of dexamethasone whereas the un-induced cells were negative and very less mineralization was observed. no difference in protein profile was observed in induced and un-induced cells by sds-page. mscs were successfully differentiated into osteoblasts even in the absence of dexamethasone. dexamethasone treatment however accelerates the cell proliferation and induces early differentiation of osteoblasts. copyright 2012 zoological society of pakistan.