Journal
STEM CELL REPORTS
Volume 6, Issue 1, Pages 18-25Publisher
CELL PRESS
DOI: 10.1016/j.stemcr.2015.11.010
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Funding
- MEXT of Japan
- Kyoto-Funding for Innovation in Health-related R&D Fields of Kyoto City
- Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Agency
- Uehara Memorial Foundation
- Grants-in-Aid for Scientific Research [15K21114, 16K15457, 15H02537] Funding Source: KAKEN
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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 NKX21(+)-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.
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