بر روي سلولهاي بنيادي مزانشيمي در محيط كشت )MAPKI( MITOGEN ACTIVATED PROTEIN KINASE INHIBITOR بررسي اثر فاكتور

Studying the effect of Mitogen Activated Protein Kinase Inhibitor (MAPKI) on Mesenchymal Stem Cells in vitro

OBJECTIVES: Multipotent mesenchymal stem cells (MSc) are known for a long period of time. These cells are adherent, clonogenic and fibroblastic and can be isolated from bone marrow stroma of postnatal organisms. Under appropriate conditions, these cells can give rise to the broad spectrum of fully differentiated connective tissues including cartilage, bone, adipose tissues and muscle cells. Upon isolating of cells in vitro, these cells tend to differentiate and it is very difficult to direct these cells into self-renewal in order to get more Multipotent MSc for medical purposes. In the previous study, the effect of leukemia inhibitory factor (L1F) on MSc was investigated and it was shown that in the presence of LIF, MSc have more proliferation ability and the self renewal ability of cells increases. Also it was shown that these cells tend to differentiate into bones. In this study the effect of Mitogen Activated Protein Kinase Inhibitor (MAPKI) on MSc was investigated. METHODS: Bone marrow stroma cells were extracted from 2-month-old mice. These cells were cultured in a medium containing 25 micro moles MAPKI. After formation of colonies, cells were stained by methylen blue. The activity of alkaline Phosphatase was also investigated. To study the effect of both LEF and MAPKI simultaneously on colony formation ability of MSc, cells were cultured in a culture containing 25 micro moles MAPKI and 500 U/ml LIF. The number and size of colonies was studied. Results: In the presence of MAPKI the number and the size of colonies (CFU-F) formed in the culture was significantly decreased. The number of CFU-ALPʹ in the culture containing MAPKI was decreased according to the decrease of CFU-F number. Adding MAPKI to a medium containing LIF neutralized the effect of LIF on MSc. Conclusion: The presence of MAPKI in the culture affects the number of colonies. This factor, despite LIF, does not affect the expression of alkaline phosphatase in Mesenchymal Stem Cells. LIF increases the self renewal and proliferative ability of cells. Adding MAPKI factor to a culture containing LIF neutralizes the proliferative effect of LIF.