Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds
Simple and Robust Differentiation of Human Pluripotent Stem Cells toward Chondrocytes by Two Small-Molecule Compounds
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DOI：
10.1016/j.stemcr.2019.07.012
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发表时间：
2019-09-10
影响因子：
5.9
通讯作者：
Saito, Taku
中科院分区：
文献类型：
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作者：
Kawata, Manabu;Mori, Daisuke;Saito, Taku
使用小分子化合物区分软骨细胞和多能干细胞（PSC）的简单诱导方案有利于软骨再生医学和软骨形成的机制研究。在这里，我们证明，通过两种化合物的简单组合，CHIR99021（一种糖原合成酶激酶 3 抑制剂）和 TTNPB（一种视黄酸受体 (RAR) 激动剂），在无血清和饲养条件下在 5-9 天内从人 PSC 中强烈诱导软骨细胞。证明了其优异的分化效率和在体内形成透明软骨组织的潜力。每个方案阶段的全面基因表达和开放染色质分析揭示了向软骨细胞的逐步分化。 RAR 和 β-连环蛋白与 DNA 关联的全基因组分析表明，视黄酸和 Wnt/β-连环蛋白信号协同调节每个分化阶段的关键标记基因。该方法为再生医学提供了一种有前途的细胞来源，并且作为体外模型，可能有助于阐明软骨细胞分化的分子机制。
A simple induction protocol to differentiate chondrocytes from pluripotent stem cells (PSCs) using small-molecule compounds is beneficial for cartilage regenerative medicine and mechanistic studies of chondrogenesis. Here, we demonstrate that chondrocytes are robustly induced from human PSCs by simple combination of two compounds, CHIR99021, a glycogen synthase kinase 3 inhibitor, and TTNPB, a retinoic acid receptor (RAR) agonist, under serum- and feeder-free conditions within 5-9 days. An excellent differentiation efficiency and potential to form hyaline cartilaginous tissues in vivo were demonstrated. Comprehensive gene expression and open chromatin analyses at each protocol stage revealed step-by-step differentiation toward chondrocytes. Genome-wide analysis of RAR and beta-catenin association with DNA showed that retinoic acid and Wnt/beta-catenin signaling collaboratively regulated the key marker genes at each differentiation stage. This method provides a promising cell source for regenerative medicine and, as an in vitro model, may facilitate elucidation of the molecular mechanisms underlying chondrocyte differentiation.