Characterization and Expression of Senescence Marker in Prolonged Passages of Rat Bone Marrow-Derived Mesenchymal Stem Cells

Figure 1

Generation and optimization of rBM-MSCs culture. Bone marrow was harvested from femur and tibia of SD rats and nucleated cells were cultured in T25 flask in day 0. By day 3, cells began to attach and heterogeneous population with predominant fibroblast-like morphology were observed by day 7 (a). One million of nucleated cells from bone marrow were cultured for 10 days in respective media and FBS concentrations. Colonies were subjected to crystal violet staining and colonies which brightly stained were counted (b). Four different basal media with 10% and 20% FBS concentration were utilized to culture 1 × 10⁶ freshly isolated BM nucleated cells for CFU and proliferation assays. CFU-f and proliferation assays were measured using crystal violet staining and trypan blue exclusion test, respectively. Results were representative of three independent experiments. ignificant values were compared between LDMEM 20% and other basal media with 20% FBS; ignificant values were compared between 10% FBS and 20% FBS of the respective media. . Microscopic magnification: 200x.

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