The present invention is directed to an agonist of NOD2 for use in therapy for increasing the autonomous capacity of survival of vertebrate adult stem cells, without loss of their capacity to multiply and differentiate, and preferably the capacity of survival of intestinal stem cells, especially in response to a stress. The invention also concerns the use of an agonist of Nod2 for increasing in vitro or ex vivo the autonomous capacity of survival, without loss of multiplication and differentiation capacity of mammalian adult stem cell. The invention also discloses different media and support for mammalian adult stem cells. The invention also concerns an in vitro screening process for identifying molecules capable increasing, in response to a stress, the autonomous capacity of survival, without loss of multiplication and differentiation capacity of mammalian adult stem cells.

Disclosed herein are cell cultures comprising dorsal and/or ventral PDX1-positive foregut endoderm cells and methods of producing the same. Also disclosed herein are cell populations comprising substantially purified dorsal and/or ventral PDX1-positive foregut endoderm cells as well as methods for enriching, isolating and purifying dorsal and/or ventral PDX1-positive foregut endoderm cells from other cell types. Methods of identifying differentiation factors capable of promoting the differentiation of dorsal and/or ventral PDX1-positive foregut endoderm cells, are also disclosed.

Described are cell culture solutions and systems for epithelial stem cell and organoid cultures, formation of epithelial constructs and uses of the same in transplantation.

This presently disclosed subject matter relates to an in vitro cell culture comprising a cell monolayer comprising mucus producing cells and a mucus layer, and methods of making and using the same. The methods including culturing mucus producing cells on a cell support structure under conditions to establish a mucus layer on the luminal side of the cell monolayer, thereby producing a live cell construct comprising a cell monolayer comprising mucus producing cells and a mucus layer. The mucus layer can be substantially impenetrable to micro-objects, and have a thickness of about 1 micron to about 1 cm.

The present invention relates to a cell culture medium for preparing liver, gastric, pancreatic, colon or intestinal adult stem cell isolated from adult tissue, as well as for maintaining such stem cell in the undifferentiated state. The cell culture medium comprises a base medium; an ABL and SRC dual kinase inhibitor/an ABL kinase inhibitor and a SRC kinase inhibitor; a mitogenic factor; a WNT signalling pathway activator; a stimulator for NAD+ and NADP+ generation; and a cAMP/P KA pathway activator. In a particular embodiment, the ABL and SRC dual kinase inhibitor is Dasatinib; the mitogenic factor is EGF; the WNT signalling pathway activator is R-Spondin 1; the stimulator for NAD+ and NADP+ generation is nicotinamide; and the cAMP/PKA pathway activator is cholera endotoxin.

The present invention provides a cell culture for obtaining an epithelial organoid, the cell culture comprising i) epithelial stem cells, or tissue fragments comprising said epithelial stem cells, ii) a basal medium for animal or human cells, iii) a Bone Morphogenetic Protein (BMP) inhibitor, iv) a mitogenic growth factor selected from the group consisting of epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), basic fibroblast growth factor (bFGF), brain-derived neurotrophic factor (BDNF) and keratinocyte growth factor (KGF), v) at least one soluble culture enhancer, wherein said at least one culture enhancer induces correct polarization of the cells in said cell culture within the developing organoid such as a laminin/entactin complex or entactin, and vi) a Wnt agonist if said epithelial stem cells, or tissue fragments comprising said epithelial stem cells are healthy cells, wherein said at least one soluble culture enhancer in said cell culture is a laminin/entactin complex in a concentration between 0.2 mg/ml and 3.4 mg/ml. A cell culture medium, an in-vitro method for obtaining an epithelial organoid, and an epithelial organoid obtained by said method are also disclosed.

It is an object to provide a culture method which is capable of maintaining and/or culturing iPS cell-derived intestinal stem cells, while maintaining the properties of intestinal stem cells. The induced pluripotent stem cell-derived intestinal stem cell-like cells are cultured in the presence of a GSK-3β inhibitor, a histone deacetylation inhibitor, and a serum replacement, or in the presence of a GSK-3β inhibitor and a serum replacement. Preferably, the culture is carried out under conditions in which one or more compounds selected from the group consisting of an epidermal growth factor, a TGFβ receptor inhibitor and a fibroblast growth factor are further present.

The present invention relates to a culture media system that is useful for the isolation and epigenetically stable propagation of normal stem cells in culture which are derived from columnar epithelial tissues and cancer stem cells from epithelial cancers. In certain embodiments, the culture system is a feeder-free system.

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.

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.