In Vitro Stage-Specific Chondrogenesis of Mesenchymal Stem Cells Committed to Chondrocytes
In Vitro Stage-Specific Chondrogenesis of Mesenchymal Stem Cells Committed to Chondrocytes
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DOI：
10.1002/art.24265
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发表时间：
2009-02-01
影响因子：
--
通讯作者：
Deng, Win-Ping
中科院分区：
文献类型：
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作者：
Chen, Wei-Hong;Lai, Ming-Tang;Deng, Win-Ping
客观的。骨关节炎的特点是软骨稳态失衡，这种失衡可以通过基于间充质干细胞（MSC）的疗法来纠正。然而，未分化间充质干细胞的体内植入却带来了意想不到的结果。本研究的目的是建立一种对 MSC 进行软骨形成预处理的模型，作为软骨再生的更有效的临床工具。方法。共培养预处理系统用于提高人 MSC 的软骨形成潜力，并使用人 MSC 系 Kp-hMSC 与人软骨细胞系 hPi（用绿色荧光蛋白 [GFP] 标记）共培养，研究 MSC 软骨形成的详细阶段。此外，将定向的间充质干细胞接种到胶原蛋白支架中并分析其新软骨形成能力。结果。 hPi-GFP 软骨细胞与 Kp-hMSC 共培养可诱导软骨形成，如软骨形成基因表达增加和软骨形成基质积累所示，但对成骨标志物没有影响。定向间充质干细胞的软骨形成过程是由高度活化的软骨形成粘附分子和刺激的软骨发育生长因子启动的，包括转化生长因子β超家族的成员及其下游调节因子Smads，以及内皮生长因子、成纤维细胞生长因子、胰岛素样生长因子和血管内皮生长因子。此外，定向的 Kp-hMSC 在胶原支架内获得了新软骨形成潜力。结论。这些发现有助于定义软骨形成的分子标记，并更准确地描绘人类间充质干细胞软骨细胞分化过程中软骨形成的阶段。结果表明，进入软骨细胞分化的软骨祖细胞阶段的人类间充质干细胞经历了详细的软骨形成变化。这种人类间充质干细胞体外软骨形成模型代表了未来临床应用细胞移植的进步。
Objective. Osteoarthritis is characterized by an imbalance in cartilage homeostasis, which could potentially be corrected by mesenchymal stem cell (MSC)-based therapies. However, in vivo implantation of undifferentiated MSCs has led to unexpected results. This study was undertaken to establish a model for preconditioning of MSCs toward chondrogenesis as a more effective clinical tool for cartilage regeneration.Methods. A coculture preconditioning system was used to improve the chondrogenic potential of human MSCs and to study the detailed stages of chondrogenesis of MSCs, using a human MSC line, Kp-hMSC, in commitment cocultures with a human chondrocyte line, hPi (labeled with green fluorescent protein [GFP]). In addition, committed MSCs were seeded into a collagen scaffold and analyzed for their neocartilage-forming ability.Results. Coculture of hPi-GFP chondrocytes with Kp-hMSCs induced chondrogenesis, as indicated by the increased expression of chondrogenic genes and accumulation of chondrogenic matrix, but with no effect on osteogenic markers. The chondrogenic process of committed MSCs was initiated with highly activated chondrogenic adhesion molecules and stimulated cartilage developmental growth factors, including members of the transforming growth factor beta superfamily and their downstream regulators, the Smads, as well as endothelial growth factor, fibroblast growth factor, insulin-like growth factor, and vascular endothelial growth factor. Furthermore, committed Kp-hMSCs acquired neocartilage-forming potential within the collagen scaffold.Conclusion. These findings help define the molecular markers of chondrogenesis and more accurately delineate the stages of chondrogenesis during chondrocytic differentiation of human MSCs. The results indicate that human MSCs committed to the chondroprogenitor stage of chondrocytic differentiation undergo detailed chondrogenic changes. This model of in vitro chondrogenesis of human MSCs represents an advance in cell-based transplantation for future clinical use.