Stepwise strategy for generating osteoblasts from human pluripotent stem cells under fully defined xeno-free conditions with small-molecule inducers
Stepwise strategy for generating osteoblasts from human pluripotent stem cells under fully defined xeno-free conditions with small-molecule inducers
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在完全确定的无异源条件下使用小分子诱导剂从人多能干细胞生成成骨细胞的逐步策略
DOI：
10.1016/j.reth.2019.12.010
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发表时间：
2020
影响因子：
4.3
通讯作者：
Ohba Shinsuke
中科院分区：
文献类型：
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作者：
Zujur Denise;Kanke Kosuke;Onodera Shoko;Tani Shoichiro;Lai Jenny;Azuma Toshifumi;Xin Xiaonan;Lichtler Alexander C.;Rowe David W.;Saito Taku;Tanaka Sakae;Masaki Hideki;Nakauchi Hiromitsu;Chung Ung-il;Hojo Hironori;Ohba Shinsuke
临床相关的人诱导多能干细胞（hiPSC）衍生物需要有效的方案来将hiPSC分化成特定谱系。在这里，我们开发了一种完全定义的无异种策略，以在21天内将hiPSC导向成骨细胞。该策略通过连续使用小分子诱导剂的组合成功地实现了四种独立衍生的hiPSC系的成骨诱导。诱导首先产生中胚层细胞，其随后概括了主要成骨细胞基因和蛋白质的发育表达模式。重要的是，Col2.3-Cherry hiPSC在这种策略下强烈表达在体内成骨成骨细胞中观察到的樱桃荧光。此外，与三维（3D）支架相结合的方案适合于生成无异种3D成骨系统。因此，我们的策略为骨生物学研究提供了一个具有显着优势的平台，也将有助于hiPSC在骨骼再生医学中的临床应用。
Clinically relevant human induced pluripotent stem cell (hiPSC) derivatives require efficient protocols to differentiate hiPSCs into specific lineages. Here we developed a fully defined xeno-free strategy to direct hiPSCs toward osteoblasts within 21 days. The strategy successfully achieved the osteogenic induction of four independently derived hiPSC lines by a sequential use of combinations of small-molecule inducers. The induction first generated mesodermal cells, which subsequently recapitulated the developmental expression pattern of major osteoblast genes and proteins. Importantly, Col2.3-Cherry hiPSCs subjected to this strategy strongly expressed the cherry fluorescence that has been observed in bone-forming osteoblastsin vivo. Moreover, the protocol combined with a three-dimensional (3D) scaffold was suitable for the generation of a xeno-free 3D osteogenic system. Thus, our strategy offers a platform with significant advantages for bone biology studies and it will also contribute to clinical applications of hiPSCs to skeletal regenerative medicine.