كشت و تكثير سلولهاي بنيادي مزانشيمي موش صحرايي با استفاده از سرم تهيه شده از خون محيطي

Culture and Expansion of Rat Mesenchymal Stem Cells Using the Serum Prepared from Ratʹs Peripheral Blood

Purpose: The aim of this study is to evaluate the effects of prepared serum from rat peripheral blood on growth and viability of rat bone marrow-derived mesenchymal stem cells (MSCs) in vitro. Materials and Methods: Bone marrow cells prepared from ratʹs tibia and femur central canal were cultured either in medium containing fetal bovin serum (FBS) or rat peripheral blood-derived serum (PBDS) for three successive passages during which viability, growth ana proliferation of the cells were evaluated by MTT [3- (4, 5- dimethylthiazol-2-yl)-2, 5- diphernyltetrazolium bromide], coiony forming assay, calculation of population doubling number and daily examination of the cell growth for drawing growth curve. Each experiment was performed ten times and the mean values were statistically compared. In this study, third passaged cells were evaluated in terms of their bone and adipocyte differentiation potentials. Results: Morphologically, the cells cultured with PBDS appeared to be somewhat more spindle than of FBS cells. Comparatively, these cells seemed to have clear cut border than of FBS-cultured cells. According to MTT assay, the cells from PBDS group significantly showed more absorption value than of FBS group indicating that they would be more viable than FBS cells. Furthermore, in this group, more colon were formed and more population doubling number (about twice of FBS cells) occurred (p<0.004). Growth curve of two cells revealed further differences; in contrast to FBS, the cell from PBDS group showed no lag phase and their log phase was much more longer than of FBS cells (2 versus 6 days). Third Passaged cells were readily differentiated to bone and adipocyte lineages confirming their mesenchymal stem cell nature. Conclusion: Taken together, the serum prepared from rat peripheral blood could significantly improve the viability and proliferation potential of the rat MSCs and in this term they could be considered as appropriate substitute for FBS.