| Objective:Bone marrow derived mesenchymal stem cells (MSCs) are reported can differentiate towards nucleus pulposus (NP)-like phenotype under specific cultural conditions. Transforming growth factor (TGF)-beta is the mostly used growth factor in NP-like differentiation of MSCs currently and other growth factors such as fibroblast growth factor (FGF)-2 is also being studied widely. However, the effects of FGF-2 on MSCs and the cocktail effects of TGF-beta and FGF-2 remains unclear, therefore, we design this study aiming at making it clarified.Methods:Human MSCs were cultured in conditioned medium contained FGF-2 or TGF-beta/FGF-2, and compared with TGF-beta and basal medium. The cells were harvested at day 7,14 and 21, and then CCK-8, dimethyl methylene blue (DMMB), real time-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the differences.Results:The groups with FGF-2 showed increased cell proliferation, especially together with TGF-beta. Functional gene markers and novel NP markers decreased both in FGF-2 group and TGF-beta group, together with functional protein expression. We found glycosaminoglycan (GAG) synthesized less in FGF-2 additive groups. Pho-ERK1/2 and pho-Smad3 differed significantly in the two conditioned groups.Conclusions:All these results demonstrate that FGF-2 can promote MSCs proliferation, synergistically with TGF-beta. However, FGF-2 plays a negative role in cartilage homeostasis. We also demonstrate FGF-2 has no positive effect in differentiating MSCs into NP-like cells, but hinders the accelerating effect of TGF-beta. |
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