A method for inducing differentiation into pancreatic α cells includes: a step (a) of culturing endodermal cells, which have been induced to differentiate from pluripotent stem cells, in the presence of a bone morphogenetic protein (BMP) signaling inhibitor, and retinoic acid or a retinoic acid analog to induce differentiation into primitive gut tube (PGT) cells; a step (b) of culturing the primitive gut tube (PGT) cells to induce differentiation into pancreatic endocrine precursor (EP) cells; and a step (c) of culturing the pancreatic endocrine precursor (EP) cells to induce differentiation into pancreatic α cells, in which the step (b) and the step (c) are performed in the absence of ascorbic acid.

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.

The present invention features, in some embodiments, compositions comprising ex vivo or in vitro generated functional enteroendocrine (EE) cells, or enteroids, rectoids, or organoids comprising functional enteroendocrine cells, and methods of obtaining and using such cells for treating metabolic and gastrointestinal diseases, disorders, or conditions.

The present invention relates to a method for preparing a human intestinal epithelial model. The human intestinal epithelial model, prepared by the method according to the present invention, has all characteristics of goblet cells, enteroendocrine cells, and Paneth cells, and thus can highly mimic the function of actual human intestinal cells, so that the human intestinal epithelial model can be effectively used for development of new drugs, evaluation of drug absorption and toxicity, or evaluation of engraftment of intestinal microorganisms, or as a composition for in vivo transplantation.

An object of the present invention is to provide a ventral hindgut organoid for producing a bladder organoid that comprises a layer structure of bladder epithelial cell types like the urinary bladder. An aspect of the present invention is to provide a method for producing a ventral hindgut organoid, comprising culturing a pluripotent stem cell with an inducer medium A containing activin A and GSK3β inhibitor to induce differentiation into definitive endoderm cells and culturing the definitive endoderm cells with an inducer medium B containing fibroblast growth factor, GSK3β inhibitor, and optionally further containing bone morphogenetic protein, and then culturing them in the presence of extracellular matrix with an inducer medium B containing fibroblast growth factor, GSK3β inhibitor, and optionally further containing bone morphogenetic protein to form a ventral hindgut organoid.

Provided are chemical inducers of pluripotency (CIP) which include glycogen synthase kinase inhibitors, TGFβ receptor inhibitors, cyclic AMP agonists and S-adenosylhomocysteine hydrolase (SAH) inhibitors or histone acetylators. A method of inducing pluripotency in a partially or completely differentiated cell by using such chemical inducers of pluripotency is also provided. The method includes: (i) contacting a cell with the CIPs for a sufficient period of time to result in reprograming the cell into a pluripotent stem cell having ESC-like characteristics (CiPSC). Isolated chemically induced pluripotent stem cells (CiPSCs) and their progeny, produced by inducing differentiation of the CiPSCs, can be used in a number of applications, including but not limited to cell therapy and tissue engineering.

Provided herein are compositions, systems, kits, and methods that employ a colitic induced human colitic organoid (iHCO) that has both an epithelial compartment and mesenchymal compartment, and provides at least one feature (e.g., leaky epithelial barrier) of IBD patient tissue (e.g., ulcerative colitis or Crohn's disease tissue). In certain embodiments, such iHCO's are employed in vitro or in vivo to screen candidate IBD treating compounds (e.g., to determine effectiveness for a particular patient who was the source of the original colonic fibroblasts used to generate the iHCO).

Provided are a small molecule compound combination for reprogramming digestive tract derived epithelial cells to endodermal stem/progenitor cells, a reprogramming method and an application. Human gastric epithelial cells (hGECs) are used as initiating cells, human gastric subepithelial myofibroblasts (aGSEMFs) are used as a trophoblast, a compound combination having all or a plurality of FBP, Bay K 8644, Bix01294, SB431542 or A813-01, VPA, RG108, PD0325901 and PS48 including SB or A83 is used to reprogram digestive tract derived epithelial cells to endodermal stem/progenitor cells, and the endodermal stem/progenitor cells can be used for inducing differentiation towards liver cells, pancreatic beta cells and intestinal cells.

A problem addressed by the present invention is to provide a method for producing pancreatic β cells from endothermal cells that have been induced to differentiate from pluripotent stein cells, wherein the method allows the production of pancreatic β cells of high quality, and to provide pancreatic progenitor (PP) cells, pancreatic endocrine precursor (EP) cells, and pancreatic β cells produced by the above-mentioned method. The present invention provides a method for producing pancreatic β 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 β cells.

The present invention relates to a combination of microbes, cell culture systems and microfluidic fluidic systems for use in providing a human Intestine On-Chip with optimal intestinal motility. More specifically, in some embodiments, a microfluidic chip containing intestinal epithelial cells co-cultured with intestinal endothelial cells in the presence of bacteria, such as probiotic bacteria, may find use in providing an Intestine-On-Chip for testing intestinal motility function. In some embodiments, an Intestine On-Chip may be used for identifying (testing) therapeutic compounds continuing probiotic microbes or compounds for inducing intestinal motility for use in treating gastrointestinal disorders or diseases related to intestinal function.