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 method for obtaining epithelial organoids is provided. In one embodiment, the method comprises culturing one or more epithelial ducts, epithelial duct fragments and/or epithelial stem cells isolated therefrom in contact with an extracellular matrix in the presence of a basal medium, wherein the medium is free of FGF and/or nicotinamide. Organoids obtained by the methods described herein, and uses thereof, are also provided.

Disclosed herein are methods involving the targeting of 5HT biosynthesis in gut insulin-negative cells to convert them into insulin-positive cells. Also, disclosed are methods for treating a disease or disorder in a mammal, preferably a human, associated with impaired pancreatic endocrine function, by administering a therapeutically effective amount of an enumerated active agent that reduces the expression, biosynthesis, signaling or biological activity of serotonin or increases its degradation, wherein administering comprises delivering the agent to Gut Ins cells in the mammal. Other embodiments of the method are directed to therapy wherein an agent that significantly reduces FOXO1 expression, biosynthesis, signaling or biological activity or increases its degradation is administered in addition to the agent that reduces serotonin, or alternatively an agent that reduces FOXO1 expression is targeted to serotonin-positive gut enteroendocrine cells.

The current invention provides for methods of promoting differentiation of human pluripotent stem cells into esophageal progenitor cells as well as the cells obtained from the methods, solutions, compositions, and pharmaceutical compositions comprising such cells. The current invention also provides for methods of using the esophageal progenitor cells for treatment and prevention of disease, and kits.

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

The technical result of the invention is to simplify the technology of obtaining insulin-producing cells, obtaining at least 70% of functionally active insulin-producing cells in cell culture, that underwent differentiation. The method comprises obtaining epithelial progenitor cells and their subsequent differentiation into pancreatic cells, capable to glucose-sensitive insulin secretion in which pancreatic differentiation is performed in two stages: (a) at the first stage cells are differentiated within 4-15 days in a culture medium containing at least serum of a mammal, glutamine, epidermal growth factor, transferrin, sodium selenite, retinoic acid, isoproterenol; (b) at the second stage, cells are differentiated within 4-15 days in a culture medium containing at least serum of a mammal, glutamine, epidermal growth factor, retinoic acid, nicotinamide, hepatocyte growth factor, dexamethasone; moreover, the cultivation in both stages is carried out in gas atmosphere of 5% CO2 at 37 C. Group of inventions includes cell product of insulin-producing cells of a mammal, and a method of differentiation of pancreatic epithelial progenitor cells of mammals, including humans, as well as a method for replacement therapy of diabetes mellitus using cell product.

Disclosed herein are methods involving the targeting of 5HT biosynthesis in gut insulin-negative cells to convert them into insulin-positive cells. Also disclosed are methods for treating a disease or disorder in a mammal, preferably a human, associated with impaired pancreatic endocrine function, by administering a therapeutically effective amount of an enumerated active agent that reduces the expression, biosynthesis, signaling or biological activity of serotonin or increases its degradation, wherein administering comprises delivering the agent to Gut Ins cells in the mammal. Other embodiments of the method are directed to therapy wherein an agent that significantly reduces FOXO1 expression, biosynthesis, signaling or biological activity or increases its degradation is administered in addition to the agent that reduces serotonin, or alternatively an agent that reduces FOXO1 expression is targeted to serotonin-positive gut enteroendocrine cells.

Provided are chemical inducers of pluripotency (CIP) which include glycogen synthase kinase inhibitors, TGF? 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 reprogramming 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.

The current invention provides for methods of promoting differentiation of human pluripotent stem cells into esophageal progenitor cells as well as the cells obtained from the methods, solutions, compositions, and pharmaceutical compositions comprising such cells. The current invention also provides for methods of using the esophageal progenitor cells for treatment and prevention of disease, and kits.

The invention relates to culture methods, in particular methods of obtaining organoid-derived monolayers, and to uses of the organoid-derived monolayers obtained by said methods. The invention also relates to assays for epithelial barrier function and methods of screening compounds using said assays.