The study describes a robust and efficient method to direct the differentiation of human pluripotent stem cells (PSCs) into intestinal tissue in vitro. The process involves a temporal series of growth factor manipulations to mimic embryonic intestinal development, including activin-induced definitive endoderm formation, FGF7/Wnt-induced posterior endoderm patterning, hindgut specification, and morphogenesis. The resulting three-dimensional intestinal "organoids" consist of a polarized, columnar epithelium with villus-like structures and crypt-like proliferative zones that express intestinal stem cell markers. The epithelium contains functional enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. The study also identifies the combined activity of Wnt3a and FGF4 as essential for hindgut specification, and NEUROG3 as both necessary and sufficient for human enteroendocrine cell development. This method allows for the generation of human intestinal tissue with a three-dimensional architecture and cellular composition similar to the fetal intestine, facilitating studies on human intestinal development and disease.The study describes a robust and efficient method to direct the differentiation of human pluripotent stem cells (PSCs) into intestinal tissue in vitro. The process involves a temporal series of growth factor manipulations to mimic embryonic intestinal development, including activin-induced definitive endoderm formation, FGF7/Wnt-induced posterior endoderm patterning, hindgut specification, and morphogenesis. The resulting three-dimensional intestinal "organoids" consist of a polarized, columnar epithelium with villus-like structures and crypt-like proliferative zones that express intestinal stem cell markers. The epithelium contains functional enterocytes, goblet cells, Paneth cells, and enteroendocrine cells. The study also identifies the combined activity of Wnt3a and FGF4 as essential for hindgut specification, and NEUROG3 as both necessary and sufficient for human enteroendocrine cell development. This method allows for the generation of human intestinal tissue with a three-dimensional architecture and cellular composition similar to the fetal intestine, facilitating studies on human intestinal development and disease.