Sensory & Motor System Medicine, Faculty of Medicine, The University of Tokyo, Tokyo, Japan; Bone and Cartilage Regenerative Medicine, Faculty of Medicine, The University of Tokyo, Tokyo, Japan.
Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo, Tokyo, Japan.
Stem Cell Reports. 2019 Sep 10;13(3):530-544. doi: 10.1016/j.stemcr.2019.07.012. Epub 2019 Aug 8.
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 β-catenin association with DNA showed that retinoic acid and Wnt/β-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.
一种使用小分子化合物将多能干细胞（PSCs）分化为软骨细胞的简单诱导方案有益于软骨再生医学和软骨发生的机制研究。在这里，我们证明了在无血清和无饲养条件下，两种化合物 CHIR99021（糖原合酶激酶 3 抑制剂）和 TTNPB（视黄酸受体（RAR）激动剂）的简单组合可在 5-9 天内从人 PSCs 中快速诱导产生软骨细胞。该方案具有良好的分化效率和在体内形成透明软骨组织的潜力。在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逐步分化的过程。对 RAR 和 β-连环蛋白与 DNA 的全基因组分析表明，视黄酸和 Wnt/β-连环蛋白信号协同调控各分化阶段的关键标记基因。该方法为再生医学提供了有前途的细胞来源，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。
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