Methods of promoting esophageal differentiation of pluripotent stem cells

A method for inducing esophageal differentiation of pluripotent stem cells, comprising the steps of

• a. differentiating the pluripotent stem cells into endoderm cells by culturing the pluripotent stem cells in an endoderm differentiation medium for about one day to about four days;
• b. differentiating the endoderm cells produced from step a. into anterior foregut cells by culturing the endoderm cells in an anterior foregut differentiation medium and (i) contacting or incubating the cultured endoderm cells with Noggin and SB431542 for about 24 hours, and (ii) further contacting or incubating the cultured endoderm cells with SB431542 and IWP-2 and wherein step b(ii) does not comprise contacting or incubating the cultured endoderm cells with Noggin for about 24 hours;
• c. further culturing the anterior foregut cells produced from step b. in a first esophageal differentiation medium for about two days to ten days, wherein the first esophageal differentiation medium comprises Noggin and SB431542;
• and
• d. further culturing the cells produced from step c in a second esophageal progenitor cell differentiation medium under conditions and for a time sufficient to allow the cells produced by step c to differentiate into esophageal progenitor cells, wherein the esophageal progenitor cells are SOX2+p63+EPCAM+ITGβ+NKX2.1-.