The invention disclosed herein generally relates to methods and systems for converting stem cells into specific tissue(s) or organ(s) through directed differentiation. In particular, the invention disclosed herein relates to methods and systems for promoting vascularized human intestinal organoid tissue having an intestine-specific EC transcriptional signature from hindgut spheroid tissue produced in vitro from the described methods.

A problem addressed by the present invention is to provide a method for producing pancreatic cells from endothermal cells that have been induced to differentiate from pluripotent stem cells, wherein the method allows the production of pancreatic cells of high quality, and to provide pancreatic progenitor (PP) cells, pancreatic endocrine precursor (EP) cells, and pancreatic cells produced by the above-mentioned method. The present invention provides a method for producing pancreatic cells that includes culturing primitive gut tube (PGT) cells, which have been induced to differentiate from pluripotent stem cells, in the presence of a protein kinase C (PKC) activator, thereby producing posterior foregut (PFG) cells; next culturing the cells in the presence of retinoic acid or an analog thereof, thereby producing pancreatic progenitor (PP) cells; next culturing the cells in the presence of a Notch signaling inhibitor and a ROCK signaling inhibitor, thereby producing pancreatic endocrine precursor (EP) cells; and next culturing the cells in the presence of an insulin receptor signaling activator, transferrin, and selenous acid, thereby producing pancreatic cells.

The invention relates to methods for developing and maintaining organoids and the organoids produced thereby.

A feeder cell for maintaining or proliferating, by a two-dimensional culture, a cell deemed to be difficult to maintain and proliferate by a two-dimensional culture; and a method of producing the feeder cell. A fibroblast-like cell is produced from a pluripotent stem cell, and derived from an intestinal structure containing an endoderm-derived cell and a mesoderm-derived cell is proliferated, isolated, and used as a feeder cell.