Method of producing airway epithelial cells

A method for producing airway epithelial cells from mammalian pluripotent stem cells comprising Steps (1) to (6) below:

• (1) culturing mammalian pluripotent stem cells in a medium containing activin A and a GSK3β inhibitor;
• (2) culturing the cells obtained in Step (1) in a medium containing a BMP inhibitor and a TGFβ inhibitor;
• (3) culturing the cells obtained in Step (2) in a medium containing BMP4, retinoic acid, and a GSK3β inhibitor to obtain ventral anterior foregut cells;
• (4) optionally culturing the ventral anterior foregut cells obtained in Step (3) in a medium containing a GSK3β inhibitor and FGF10 to induce the cells to differentiate into airway epithelial progenitor cells;
• (5) subjecting the ventral anterior foregut cells obtained in Step (3) or the airway epithelial progenitor cells obtained in Step (4) to three-dimensional culture in a medium containing a GSK3β inhibitor, FGF10, and a ROCK inhibitor to obtain proximal airway epithelial progenitor cells;
• and
• (6) subjecting the proximal airway epithelial progenitor cells obtained in Step (5) to three-dimensional culture in a medium containing a ROCK inhibitor but not containing a GSK3β inhibitor, thereby producing airway epithelial cells;
• wherein the airway epithelial cells are selected from the group consisting of ciliated airway epithelial cells, airway mucin-producing cells, basal airway epithelial cells, and Club cells.