Method for inducing differentiation of alveolar epithelial cells

A method for producing a cell population comprising type II alveolar epithelial cells from pluripotent stem cells comprising Steps (1) to (5) below:

• (1) culturing pluripotent stem cells in a medium containing activin A and a glycogen synthase kinase 3β (GSK3β) inhibitor;
• (2) culturing the cells obtained in Step (1) in a medium containing a morphogenetic protein (BMP) inhibitor and a transforming growth factor beta (TGFβ) inhibitor;
• (3) culturing the cells obtained in Step (2) in a medium containing bone morphogenetic protein 4 (BMP4), retinoic acid, and a GSK3β inhibitor;
• (4) culturing the ventral anterior foregut cells obtained in Step (3) in a medium containing a GSK3β inhibitor, Fibroblast Growth Factor 10 (FGF10), keratinocyte growth factor (KGF), and a NOTCH signal inhibitor for a duration of time until alveolar epithelial progenitor cells are induced;
• followed by a step of isolating carboxypeptidase M-positive (CPM-positive) cells as alveolar epithelial progenitor cells;
• and
• (5) subjecting the induced alveolar epithelial progenitor cells obtained in Step (4) to three-dimensional culture in a basal medium supplemented with additives consisting of a steroid drug, a cAMP derivative, a phosphodiesterase inhibitor, and KGF;
• following Step (5), a further step of isolating cells positive for one or more type II alveolar epithelial cell markers selected from the group consisting of surfactant protein C (SFTPC), epithelial cell adhesion molecule (EpCAM), and carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) as type II alveolar epithelial cells;
• wherein cells positive for staining of acidic fractions for live cells are isolated as type II alveolar epithelial cells;
• thereby obtaining a cell population wherein the type II alveolar epithelial cells are at least 50% of the cell population, relative to total epithelial cells.