Sequential exposure to fibroblast growth factors (FGF) 2, 9 and 18 enhances hMSC chondrogenic differentiation.
Sequential exposure to fibroblast growth factors (FGF) 2, 9 and 18 enhances hMSC chondrogenic differentiation.
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DOI：
10.1016/j.joca.2014.11.013
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发表时间：
2015-03
影响因子：
7
通讯作者：
Caplan AI
中科院分区：
文献类型：
--
作者：
Correa D;Somoza RA;Lin P;Greenberg S;Rom E;Duesler L;Welter JF;Yayon A;Caplan AI
检测体外软骨形成过程中连续暴露于FGF 2、9和18对人间充质干细胞（hMSC）分化的影响。在软骨诱导程序期间的不同时间开始，在rhFGF 9、rhFGF 18或rhFGFR 3特异性信号转导FGF变体的存在下，将对照和FGF 2扩增的hMSC培养在聚集体中。在不同阶段进行qRT-PCR和免疫细胞化学。将聚集体培养物与rhFGF和抗FGFR 1和FGFR 3的中和抗体一起沿着转移至肥大诱导培养基中。进行组织学/免疫组织化学/生化分析。在扩增过程中暴露于FGF 2的hMSC上调Sox 9，表明软骨形成机制的早期激活。FGF 2、FGF 9和18在hMSC扩增和软骨形成过程中调节FGFR 1和FGFR 3的表达谱。与TGF-β组合，当在程序开始时（≤ d 7）添加时，FGF 9和FGF 18抑制软骨形成，而当稍后（≥d14）添加时，表现出合成代谢作用，该作用由FGFR 3介导。最后，由FGF 9或FGF 18诱导的FGFR 3信号传导延迟了自发性和诱导的肥大相关变化的出现。加入生长因子的hMSC依赖性软骨形成阶段影响分化程序的进展：增加细胞增殖和引发（FGF 2）;通过更早地改变软骨形成程序刺激早期软骨形成分化（TGF-β，FGF 9/FGF 18）;增加ECM产生（FGF 9/FGF 18）;和延迟终末肥大（FGF 9/FGF 18）。总的来说，这些因素可以用来优化植入前条件的hMSC时，用于工程软骨移植。
To test the effects of sequential exposure to FGF2, 9 and 18 on human Mesenchymal Stem Cells (hMSC) differentiation during in vitro chondrogenesis. Control and FGF2-expanded hMSC were cultured in aggregates in the presence of rhFGF9, rhFGF18 or rhFGFR3-specific signaling FGF variants, starting at different times during the chondroinductive program. qRT-PCR and immunocytochemistry were performed at different stages. The aggregate cultures were switched to a hypertrophy-inducing medium along with rhFGFs and neutralizing antibodies against FGFR1 and FGFR3. Histological/immunohistochemical/biochemical analyses were performed. FGF2-exposed hMSC during expansion up-regulated Sox9 suggesting an early activation of the chondrogenic machinery. FGF2, FGF9 and 18 modulated the expression profile of FGFR1 and FGFR3 in hMSC during expansion and chondrogenesis. In combination with TGF-β, FGF9 and FGF18 inhibited chondrogenesis when added at the beginning of the program (≤d7), while exhibiting an anabolic effect when added later (≥d14), an effect mediated by FGFR3. Finally, FGFR3 signaling induced by either FGF9 or FGF18 delayed the appearance of spontaneous and induced hypertrophy-related changes. The stage of hMSC-dependent chondrogenesis at which the growth factors are added impacts the progression of the differentiation program: increased cell proliferation and priming (FGF2); stimulated early chondrogenic differentiation (TGF-β, FGF9/FGF18) by shifting the chondrogenic program earlier; augmented ECM production (FGF9/FGF18); and delayed terminal hypertrophy (FGF9/FGF18). Collectively, these factors could be used to optimize pre-implantation conditions of hMSC when used to engineer cartilage grafts.