Zujur Denise, Al-Akashi Ziadoon, Nakamura Anna, Zhao Chengzhu, Takahashi Kazuma, Aritomi Shizuka, Theoputra William, Kamiya Daisuke, Nakayama Koichi, Ikeya Makoto
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Center for Regenerative Medicine Research, Faculty of Medicine, Saga University, Saga, Japan.
Front Cell Dev Biol. 2023 May 10;11:1140717. doi: 10.3389/fcell.2023.1140717. eCollection 2023.
To date, there is no effective long-lasting treatment for cartilage tissue repair. Primary chondrocytes and mesenchymal stem/stromal cells are the most commonly used cell sources in regenerative medicine. However, both cell types have limitations, such as dedifferentiation, donor morbidity, and limited expansion. Here, we report a stepwise differentiation method to generate matrix-rich cartilage spheroids from induced pluripotent stem cell-derived mesenchymal stem/stromal cells (iMSCs) via the induction of neural crest cells under xeno-free conditions. The genes and signaling pathways regulating the chondrogenic susceptibility of iMSCs generated under different conditions were studied. Enhanced chondrogenic differentiation was achieved using a combination of growth factors and small-molecule inducers. We demonstrated that the use of a thienoindazole derivative, TD-198946, synergistically improves chondrogenesis in iMSCs. The proposed strategy produced controlled-size spheroids and increased cartilage extracellular matrix production with no signs of dedifferentiation, fibrotic cartilage formation, or hypertrophy . These findings provide a novel cell source for stem cell-based cartilage repair. Furthermore, since chondrogenic spheroids have the potential to fuse within a few days, they can be used as building blocks for biofabrication of larger cartilage tissues using technologies such as the Kenzan Bioprinting method.
迄今为止,对于软骨组织修复尚无有效的长效治疗方法。原代软骨细胞和间充质干/基质细胞是再生医学中最常用的细胞来源。然而,这两种细胞类型都有局限性,如去分化、供体发病以及扩增受限。在此,我们报告一种逐步分化方法,在无血清条件下通过诱导神经嵴细胞,从诱导多能干细胞衍生的间充质干/基质细胞(iMSCs)生成富含基质的软骨球体。研究了在不同条件下生成的iMSCs软骨形成敏感性的调控基因和信号通路。使用生长因子和小分子诱导剂的组合实现了增强的软骨形成分化。我们证明,噻吩并吲唑衍生物TD - 198946的使用可协同改善iMSCs的软骨形成。所提出的策略产生了大小可控的球体,并增加了软骨细胞外基质的产生,且没有去分化、纤维化软骨形成或肥大的迹象。这些发现为基于干细胞的软骨修复提供了一种新的细胞来源。此外,由于软骨形成球体有可能在几天内融合,它们可用作使用如Kenzan生物打印方法等技术生物制造更大软骨组织的构建模块。