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The mRNA MPEP levels of p16INK4a, p21Cip1, and p53 and the expression levels of pRB in hMSCs were measured after culture for 10 or 50 days in FGF-2(?) or FGF-2(+). After 50 days’ culture in FGF-2(?), the mRNA expression levels of p16INK4a, p21Cip1, and p53 were significantly higher than after 10 days’ culture, but not after culture in FGF-2(+) (Fig. 3A–C). On the other hand, after 50 days’ culture in FGF-2(?), the expression levels of total pRB and ppRB were decreased compared with after 10 days’ culture, but not after culture in FGF-2(+) (Fig. 3D). These results suggest that FGF-2 suppresses hMSCs cellular senescence depending on the length of culture.
Fig. 3.
FGF-2 suppresses G1 cell cycle arrest due to passaging. hMSCs were maintained in the medium in the presence or absence of FGF-2 (1 ng/ml), and total RNAs and proteins were extracted when approaching confluence. (A–C) p16INK4a (A), p53 (B), and p21Cip1 (C) mRNA expression levels were measured using real time RT-PCR at Days 10 and 50. (D) Total pRB and phospho-pRB proteins detected using Western blot analysis at Days 10 and 50.