Fibroblast growth factor-2 primes human mesenchymal stem cells for enhanced chondrogenesis.
Fibroblast growth factor-2 primes human mesenchymal stem cells for enhanced chondrogenesis.
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DOI：
10.1371/journal.pone.0022887
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Li WJ
中科院分区：
文献类型：
--
作者：
Handorf AM;Li WJ
人间充质干细胞（hMSCs）是一种多能细胞，能够分化为多种成熟细胞类型，包括成骨细胞、脂肪细胞和软骨细胞。先前的研究表明，当在添加成纤维细胞生长因子-2 （FGF-2）的培养基中扩增时，hMSCs显示出增强的软骨形成（CG）。先前的研究表明，CG的增强可能归因于具有固有软骨形成潜力的细胞亚群的选择。在这项研究中，我们发现FGF-2预处理实际上通过增加基础Sox9蛋白水平来启动hMSCs进行增强的CG。我们的研究结果表明，Sox9蛋白水平在暴露于FGF-2 30分钟内升高，并随着暴露时间的延长而逐渐升高。此外，我们使用流式细胞术显示，FGF-2增加了增殖和非增殖hMSCs中每个细胞的Sox9蛋白水平，这强烈表明FGF-2通过调节Sox9启动hMSCs进行后续CG。事实上，当hMSCs暴露于FGF-2 2小时后，随后通过颗粒培养分化为软骨细胞谱系时，磷酸化sox9 （pSox9）蛋白水平升高，最终导致CG增强。然而，在hMSC扩增过程中，小干扰RNA （small interfering RNA, siRNA）介导的Sox9的敲低并不能否定FGF-2的促软骨作用，这表明FGF-2介导的hMSC CG增强仅部分通过Sox9调节。我们的研究结果为FGF-2调节预分化hMSCs进行增强CG的机制提供了新的见解。
Human mesenchymal stem cells (hMSCs) are multipotent cells capable of differentiating into a variety of mature cell types, including osteoblasts, adipocytes and chondrocytes. It has previously been shown that, when expanded in medium supplemented with fibroblast growth factor-2 (FGF-2), hMSCs show enhanced chondrogenesis (CG). Previous work concluded that the enhancement of CG could be attributed to the selection of a cell subpopulation with inherent chondrogenic potential. In this study, we show that FGF-2 pretreatment actually primed hMSCs to undergo enhanced CG by increasing basal Sox9 protein levels. Our results show that Sox9 protein levels were elevated within 30 minutes of exposure to FGF-2 and progressively increased with longer exposures. Further, we show using flow cytometry that FGF-2 increased Sox9 protein levels per cell in proliferating and non-proliferating hMSCs, strongly suggesting that FGF-2 primes hMSCs for subsequent CG by regulating Sox9. Indeed, when hMSCs were exposed to FGF-2 for 2 hours and subsequently differentiated into the chondrogenic lineage using pellet culture, phosphorylated-Sox9 (pSox9) protein levels became elevated and ultimately resulted in an enhancement of CG. However, small interfering RNA (siRNA)-mediated knockdown of Sox9 during hMSC expansion was unable to negate the prochondrogenic effects of FGF-2, suggesting that the FGF-2-mediated enhancement of hMSC CG is only partly regulated through Sox9. Our findings provide new insights into the mechanism by which FGF-2 regulates predifferentiation hMSCs to undergo enhanced CG.
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