The combination of SOX5, SOX6, and SOX9 (the SOX trio) provides signals sufficient for induction of permanent cartilage
The combination of SOX5, SOX6, and SOX9 (the SOX trio) provides signals sufficient for induction of permanent cartilage
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DOI：
10.1002/art.20611
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发表时间：
2004-11-01
影响因子：
--
通讯作者：
Chung, UI
中科院分区：
文献类型：
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作者：
Ikeda, T;Kamekura, S;Chung, UI
Objective.为了再生永久性软骨，不仅要知道软骨形成的必要条件，而且要知道充分条件。的目标。本研究是为了确定足以用于软骨形成的信号。用软骨形成所必需的因子的组合处理已经被工程化以在软骨细胞分化时发荧光的胚胎干细胞，并且软骨细胞分化被检测为荧光。我们筛选了可以在3天内诱导荧光的组合。然后，原代间充质干细胞，nonchondrogenic永生化细胞系，和原代真皮成纤维细胞的组合处理，软骨细胞分化的诱导进行了评估，通过检测软骨标记基因的表达和蛋白多糖丰富的基质的积累。比较了单层、球状体和三维培养系统对转录因子组合诱导的影响。通过检测特征性标志基因的表达来评价联合用药对肥大细胞和成骨细胞分化的影响。无单因素诱导荧光。在检测的各种组合中，只有SOX 5、SOX 6和SOX 9组合（SOX三重奏）在3天内诱导荧光。SOX三人组成功地诱导了所有测试细胞类型（包括非软骨形成类型）中的软骨细胞分化，并且无论使用何种培养系统，诱导都发生了。与传统的软骨形成技术相反，SOX三联体同时抑制了肥大和成骨分化。这些数据有力地表明SOX三联体提供了足以诱导永久性软骨的信号。
Objective. To regenerate permanent cartilage, it is crucial to know not only the necessary conditions for chondrogenesis, but also the sufficient conditions. The objective of. this study was to determine the signal sufficient for chondrogenesis.Methods. Embryonic stem cells that had been engineered to fluoresce upon chondrocyte differentiation were treated with combinations of factors necessary for chondrogenesis, and chondrocyte differentiation was detected as fluorescence. We screened for the combination that could induce fluorescence within 3 days. Then, primary mesenchymal stem cells, nonchondrogenic immortalized cell lines, and primary dermal fibroblasts were treated with the combination, and the induction of chondrocyte differentiation was assessed by detecting the expression of the cartilage marker genes and the accumulation of proteoglycan-rich matrix. The effects of monolayer, spheroid, and 3-dimensional culture systems on induction by combinations of transcription factors were compared. The effects of the combination on hypertrophic and osteoblastic differentiation were evaluated by detecting the expression of the characteristic marker genes.Results. No single factor induced fluorescence. Among various combinations examined, only the SOX5, SOX6, and SOX9 combination (the SOX trio) induced fluorescence within 3 days. The SOX trio successfully induced chondrocyte differentiation in all cell types tested, including nonchondrogenic types, and the induction occurred regardless of the culture system used. Contrary to the conventional chondrogenic techniques, the SOX trio suppressed hypertrophic and osteogenic differentiation at the same time.Conclusion. These data strongly suggest that the SOX trio provides signals sufficient for the induction of permanent cartilage.