Figure 1
MM-BMMSCs exhibit a higher senescence state than HD-BMMSCs. Asterisks indicate p-values with * <0.05, ** < 0.01, *** < 0.001 and **** < 0.0001. All data were analyzed using Mann–Whitney U test and unpaired t-test (ELISA analysis). (A) Flow cytometric analysis of SA-βGalA. ND-MM-BMMSCs (n = 12) and R-MM-BMMSCs (n = 9) displayed higher activity of SA-βGal in passage 1 and 4 of cell cultures compared to HD-BMMSCs (n = 8). (B) Histological analysis of SA-βGalA. MM-BMMSC (59 years) showed aproximately 17 cells with high SA-βGalA compared to HD-BMMSC (67 years) with only 10 cells. Positive cells are indicated by arrows. (C) QPCR analysis displayed decreased cyclin E1, increased cyclin D1 and p21 expression in ND-MM-BMMSCs (n = 24) and R-MM-BMMSCs (n = 21) compared to HD-BMMSCs (n = 10). (D) Measurement of protein level in HD-BMMSCs (n = 2) and MM-BMMSCs (n = 3). Cyclin E1 was significantly decreased in MM-BMMSCs compared to HD-BMMSCs whereas cyclin D1 and p21 were increased. The protein amount of p16 was slightly reduced in MM-BMMSCs compared to HD-BMMSCs. (E) Cell cycle analysis showed a higher amount of cell in S phase in ND-MM-BMMSCs (n = 11) and R-MM-BMMSCs (n = 5) compared to HD-BMMSCs (n = 6). Percentage of cells in G1/G0 phase was lower compared to HD-BMMSCs.