In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells
In Vitro Fabrication of Hybrid Bone/Cartilage Complex Using Mouse Induced Pluripotent Stem Cells
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DOI：
10.3390/ijms21020581
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发表时间：
2020-01-02
影响因子：
5.6
通讯作者：
Egusa, Hiroshi
中科院分区：
文献类型：
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作者：
Limraksasin, Phoonsuk;Kondo, Takeru;Egusa, Hiroshi
细胞凝聚和机械刺激在成骨和软骨形成中起作用；因此，它们有望促进体外诱导多能干细胞（iPSCs）自组织骨/软骨组织的形成。在这里，首先将单个小鼠iPSCs植入微空间培养板中形成三维球体。在第12天，iPSC球进行摇培养，并在成骨诱导培养基中维持31天（Os诱导）。在另一种情况下，在第22天将成骨诱导培养基替换为软骨诱导培养基，并继续维持21天（Os-Chon诱导）。Os诱导产生了强健的矿化和一些软骨样组织，促进了成骨和软骨标记基因的表达。相反，Os-Chon诱导导致部分矿化和大面积软骨组织，软骨标记基因以及骨和胶原1a1的表达显著增加。Os-Chon诱导增强了中胚层谱系的承受性，通过近轴表达，随后高表达侧板和近轴中胚层标记基因。这些结果表明，微空间培养和机械刺激的结合使用有助于iPSC形成混合骨/软骨组织，并且可以通过诱导方案操纵iPSC构建中的骨/软骨组织比例。
Cell condensation and mechanical stimuli play roles in osteogenesis and chondrogenesis; thus, they are promising for facilitating self-organizing bone/cartilage tissue formation in vitro from induced pluripotent stem cells (iPSCs). Here, single mouse iPSCs were first seeded in micro-space culture plates to form 3-dimensional spheres. At day 12, iPSC spheres were subjected to shaking culture and maintained in osteogenic induction medium for 31 days (Os induction). In another condition, the osteogenic induction medium was 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, which promoted expression of osteogenic and chondrogenic marker genes. In contrast, Os-Chon induction resulted in partial mineralization and a large area of cartilage tissue, with greatly increased expression of chondrogenic marker genes along with osterix and collagen 1a1. Os-Chon induction enhanced mesodermal lineage commitment with brachyury expression followed by high expression of lateral plate and paraxial mesoderm marker genes. These results suggest that combined use of micro-space culture and mechanical stimuli facilitates hybrid bone/cartilage tissue formation from iPSCs, and that the bone/cartilage tissue ratio in iPSC constructs could be manipulated through the induction protocol.