Division of Molecular and Regenerative Prosthodontics, Tohoku University Graduate School of Dentistry, Sendai, Miyagi, Japan.
Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Methods Mol Biol. 2022;2454:509-520. doi: 10.1007/7651_2021_361.
In the developing embryo, bone and cartilage share the same progenitors. However, osteo-chondrogenic induction of mouse induced pluripotent stem cells (iPSCs) remains difficult. Here we describe a protocol to guide iPSCs to differentiate into osteochondral cells that form hybrid bone/cartilage constructs in vitro. Single mouse iPSCs are first reaggregated in ultra-low-attachment micro-space culture plates. At day 12, iPSC spheres are subjected to shaking culture and maintained in an osteogenic induction medium for 31 days (Os induction). In another condition, the osteogenic induction medium is replaced by chondrogenic induction medium at day 22 and maintained for a further 21 days (Os-Chon induction). Os induction produced robust mineralization and some cartilage-like tissue, whereas Os-Chon induction resulted in partial mineralization and a large area of cartilage tissue.
在发育中的胚胎中，骨骼和软骨共享相同的祖细胞。然而，诱导多能干细胞（iPSCs）向成骨-软骨诱导仍然很困难。本文描述了一种方案，可指导 iPSCs 分化为软骨细胞，从而在体外形成杂交骨/软骨构建体。首先将单个小鼠 iPSCs 在超低附着微空间培养板中重新聚集。在第 12 天，将 iPSC 球进行摇动培养，并在成骨诱导培养基中维持 31 天（Os 诱导）。在另一种条件下，在第 22 天将成骨诱导培养基替换为软骨诱导培养基，并进一步维持 21 天（Os-Chon 诱导）。Os 诱导产生了强大的矿化和一些软骨样组织，而 Os-Chon 诱导则导致部分矿化和大面积的软骨组织。
