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.

The present invention relates to a method of culturing and/or monitoring epithelial cells using a microfluidic cell culture system comprising a microfluidic channel network. In the method epithelial cells are lined, in the microfluidic cell culture system by cells of mesenchymal origin. The cells may form a tubular or tube-like structure, i.e. a.tube in a tube. The method allows for improved epithelial models suitable for a wide variety of applications, including but not limited to high-throughput screening and analysis of epithelium in health and disease.

An object of the present invention is to provide a method of producing an enteroendocrine cell by using an intestinal stem cell induced from a pluripotent stem cell, a method of analyzing a disease mechanism, an evaluation method for a test substance, a screening method for a therapeutic drug for a digestive system disease, an enteroendocrine cell derived from a pluripotent stem cell, a culture medium or a culture medium kit, and a kit for producing an enteroendocrine cell. According to the present invention, there is provided a method of producing an enteroendocrine cell, the method including an intestinal stem cell or an intestinal epithelial cell, which is induced from a pluripotent stem cell, by using a culture medium that substantially does not contain a ligand of an epidermal growth factor but contains a cAMP activator and a DNA methyltransferase inhibitor.

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.

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.

Embodiments of the present invention are directed to various methods for generating airway and lung progenitors and epithelial cells and three-dimensional anterior foregut spheres, and to populations of cells made using the methods. The airway and lung progenitors and epithelial cells can be used as a model to study diseases that primarily affect airway epithelial cells, and to study human lung development. Methods are also provided for drug screening. Anterior foregut spheres can be used as a model for lung fibrosis.

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 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.

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.

Tumor stem cells can be obtained by culturing a tumor cell population, and exposing the cultured tumor cell population to free radicals. In certain embodiments, the free radical agent can be a nitric oxide (NO) donor. In one embodiment, the free radical agent can be Diethylenetriamine NONOate (DETA NONOate) or agents that constitutively increase cellular nitric oxide, such as phosphodiesterase inhibitors or L-arginine, or agents that increase NO synthase in the population. The methods can further include inducing stem cells present in the population to expand and/or inducing dedifferentiation of tumor cells into tumor stem cells. Additionally, the present invention provides methods of selecting stem cells from a tumor cell population. Another aspect provides methods of screening for anti-tumor stem cell teherapeutic compounds by providing high nitric oxide (HNO) tumor cells, exposing the HNO cells to at least one compound, assessing one or more indicators of HNO cell health and determining toxicity of the compound to HNO tumor cells.

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.