As an undifferentiated cell, stem cell can differentiate into specialized cells (pluripotency) and can divide with mitosis to produce more stem cells (self renewal). Mesenchymal stem cells obtain in adult organisms are multipotent cells that can differentiate into a variety of cell types. They generally coordinate tissue repair mechanism. So, they use widely in tissue engineering protocols because of their pluripotency capacity. As known tissue engineering is a promising therapeutic approach to repair or regenerate damaged tissues. In our study, we aimed to obtain mouse bone marrow derived mesenchymal stem cells from bone marrow. First, bone marrow samples were obtained from mouse (mus musculus), by using long-term bone marrow culture methodology, mesenchymal stem cells were obtained. In microscope, colony forming, specific for mesenchymal stem cells were visualized. The cells in cell culture were analyzed for cell viability with trypan blue and MTT analyses in early phase of culture and late phase of culture. CD73, CD90, CD105, CD34, CD45, CD11b genes were used as genetic marker for finding the stemness properties of mesenchymal stem cells. In that step, the expression profiles of these genes were found by using real time polymerase chain reaction analyses. The cell viability and the gene expression Results of cultured cells in early phase were found higher than the Results of cultured cells in late phase. The Results represent that the methodology applied in our study is proper for having mesenchymal stem cells from bone marrow. The cultured cells have high viability and stemness properties in the early phase of cell culture. The cells lose their vitality and stemness properties in the late phase of cell culture. This finding represents that mesenchymal stem cells obtained from in the early stages of culture could be used in the advanced application of stem cells.
Keywords: Stem cells, mesenchymal stem cells, cell survival, cell surface antigen, gene expression
