Cell replacement therapy for pulmonary diseases

A method for generating lung progenitor cells, the method comprising:

• (a) producing anterior foregut endoderm cells from mammalian pluripotent stem cells (PSCs);
• (b) culturing the anterior foregut endoderm cells in a suspension culture comprising a glycogen synthase kinase (GSK) inhibitor, a bone morphogenic protein (BMP) agonist, at least one FGF agonists, and retinoic acid, to generate a lung bud organoid (LBO);
• (c) dissociating the LBO and culturing the dissociated LBO on a first cell culture substrate in presence of a GSK inhibitor, a BMP agonist, at least one FGF agonist, and retinoic acid, to generate plated LBO, wherein the first cell culture substrate is coated with a first biomolecule;
• and
• (d) culturing the plated LBO on a second cell culture substrate in a first culture medium or a second culture medium, wherein the first culture medium comprises an inhibitor of Rho kinase (ROCK), a GSK inhibitor, at least one FGF agonist, a BMP agonist, and retinoic acid, wherein the second culture medium comprises insulin, an EGF agonist, a corticosteroid, a 3′,5′-cyclic adenosine monophosphate (cAMP) pathway activator, and an inhibitor of ROCK, wherein the second cell culture substrate is coated with a second biomolecule.