Directed Induction of Functional Multi-ciliated Cells in Proximal Airway Epithelial Spheroids from Human Pluripotent Stem Cells.
Directed Induction of Functional Multi-ciliated Cells in Proximal Airway Epithelial Spheroids from Human Pluripotent Stem Cells.
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DOI：
10.1016/j.stemcr.2015.11.010
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发表时间：
2016-01-12
影响因子：
5.9
通讯作者：
Mishima M
中科院分区：
文献类型：
--
作者：
Konishi S;Gotoh S;Tateishi K;Yamamoto Y;Korogi Y;Nagasaki T;Matsumoto H;Muro S;Hirai T;Ito I;Tsukita S;Mishima M
多纤毛气道细胞（MCACs）在肺的粘液纤毛清除中发挥作用。然而，从人多能干细胞有效诱导功能性MCACs尚未报道。使用羧肽酶M（CPM）作为NKX 2 -1+腹前肠内胚层细胞（VAFECs）的表面标志物，我们报告了一种三维分化方案，用于从CPM+ VAFECs生成近端气道上皮祖细胞球状体。这些球状体可以被诱导产生MCACs和其他气道谱系细胞，而不需要肺泡上皮细胞。此外，通过添加DAPT（一种Notch途径抑制剂）促进MCACs和肺神经内分泌谱系细胞的定向诱导。诱导的MCACs表现出具有“9 + 2”微管排列的运动纤毛和能够搏动并产生用于粘液纤毛运输的流动的动力蛋白臂。该方法有望为人类气道疾病建模和再生医学的未来研究提供有用的信息。CPM是产生人近端气道上皮的有用标记物三维培养可用于体外诱导人气道上皮DAPT促进多纤毛和肺神经内分泌细胞的诱导诱导的多纤毛气道细胞具有功能性运动纤毛多纤毛气道细胞（MCACs）在粘液纤毛清除中起关键作用。Gotoh，Konishi及其同事证明，CPM+腹侧前肠内胚层细胞的三维（3D）分化可用于产生近端气道上皮，涉及功能性MCACs以及肺神经内分泌细胞（PNEC）。通过添加Notch途径抑制剂DAPT促进MCACs和PNEC的诱导。
Multi-ciliated airway cells (MCACs) play a role in mucociliary clearance of the lung. However, the efficient induction of functional MCACs from human pluripotent stem cells has not yet been reported. Using carboxypeptidase M (CPM) as a surface marker of NKX2-1+-ventralized anterior foregut endoderm cells (VAFECs), we report a three-dimensional differentiation protocol for generating proximal airway epithelial progenitor cell spheroids from CPM+ VAFECs. These spheroids could be induced to generate MCACs and other airway lineage cells without alveolar epithelial cells. Furthermore, the directed induction of MCACs and of pulmonary neuroendocrine lineage cells was promoted by adding DAPT, a Notch pathway inhibitor. The induced MCACs demonstrated motile cilia with a “9 + 2” microtubule arrangement and dynein arms capable of beating and generating flow for mucociliary transport. This method is expected to be useful for future studies on human airway disease modeling and regenerative medicine. CPM is a useful marker for generating human proximal airway epithelium Three-dimensional culture is useful for inducing human airway epithelium in vitro DAPT promotes the induction of multi-ciliated and pulmonary neuroendocrine cells Induced multi-ciliated airway cells have functionally motile cilia Multi-ciliated airway cells (MCACs) play a crucial role in mucociliary clearance. Gotoh, Konishi, and colleagues demonstrated that three-dimensional (3D) differentiation of CPM+ ventralized anterior foregut endoderm cells is useful for generating proximal airway epithelium, involving functional MCACs as well as pulmonary neuroendocrine cells (PNECs). The induction of MCACs and PNECs was promoted by adding DAPT, a Notch pathway inhibitor.