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

Figure 3

Growth kinetics and doubling time of rBM-MSC. Morphological observation of rBM-MSC cultured in LDMEM (20% FBS) in the presence of 20 ng/mL, 1% ITS, and 1% NEAA at various passages. Cells were successfully expanded until passage 5 and assumed a spindle-shaped fibroblast-like morphology. As the passage increased, polygonal and flatten shaped cells were predominating (a). Rat BM-MSCs were plated in 6-well plate at 150,000 cells/well, and media were changed every 2 days for 6 days. Cells depicted an initial lag phase for 1 day, followed by exponential log phase for 4 days, and then a plateau phase was observed (b). Doubling time was determined by Patterson Formula and expressed as mean doubling time (c). The result represented the mean ± SD. Microscopic magnification: 200x.

(a)

(b)

(c)