It is an object of the present invention to provide a method for efficiently directing differentiation into insulin-producing cells in a xeno-free culture system. According to the present invention, there is provided a method for directed differentiation into insulin-producing cells, comprising culturing stem cells in the following steps (1) to (5): (1) a step of culturing stem cells in a medium comprising an activator of activin receptor-like kinase-4/-7 and a GSK3 inhibitor and then culturing in a medium comprising an activator of activin receptor-like kinase-4/-7; (2) a step of culturing the cells obtained in step (1) in a medium comprising a hedgehog signaling inhibitor and an FGF; (3) a step of culturing the cells obtained in step (2) in a medium comprising a retinoic acid receptor agonist, a hedgehog signaling inhibitor and a BMP signaling inhibitor; (4) a step of culturing the cells obtained in step (3) in a medium comprising a TGF- type I activin receptor-like kinase-4/-5/-7 inhibitor and a BMP signaling inhibitor; and (5) a step of culturing the cells obtained in step (4) in a medium comprising a phosphodiesterase inhibitor.

The present invention relates to compositions, kits and methods for making and using RNA compositions comprising in vitro-synthesized ssRNA inducing a biological or biochemical effect in a mammalian cell or organism into which the RNA composition is repeatedly or continuously introduced. In certain embodiments, the invention provides compositions and methods for changing the state of differentiation or phenotype of a human or other vertebrate cell. For example, the present invention provides mRNA and methods for reprogramming cells that exhibit a first differentiated state or phenotype to cells that exhibit a second differentiated state or phenotype, such as to reprogram human somatic cells to pluripotent stem cells.

The present invention relates to methods of producing chondrocytes, improving the phenotype of a chondrocyte population, promoting chondrogenesis, maintaining a cell population in a differentiated state or increasing the number of cells of a population in a differentiated state, producing a population of differentiated cells, and treating certain diseases or disorders. Each of these methods involves treating a cell population with, and/or culturing a cell population in, a ceramidase

An undifferentiated stem cell-removing agent which contains at least one kind selected from the group consisting of a fatty acid synthesis inhibitor, a fatty acid utilization inhibitor, and a cholesterol synthesis inhibitor; a method for removing undifferentiated stem cells which includes culturing a cell mixture that contains an undifferentiated stem cell and a differentiated cell in the presence of the undifferentiated stem cell-removing agent; and a production method of cells for transplantation which includes the following steps (i) and (ii): a step (i) of inducing a desired differentiated cell from an undifferentiated stem cell, and a step (ii) of culturing the cell mixture obtained in the step (i) in the presence of the undifferentiated stem cell-removing agent are provided.

This invention relates to compositions and methods for activating and promoting mineralization in tissue that does not normally mineralize, specifically intervertebral discs. The composition comprises agents that increase the expression of the gene that encodes TNAP and/or the activation, amount or activity of TNAP protein, and agents that decrease the expression of ANK and/or ENPP and/or the activation, amount or activity of these proteins. The composition can be in the form of a cell or cells. The invention also relates to methods of using the composition.

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.

This invention relates to compositions and methods for activating and promoting mineralization in tissue that does not normally mineralize, specifically intervertebral discs. The composition comprises agents that increase the expression of the gene that encodes TNAP and/or the activation, amount or activity of TNAP protein, and agents that decrease the expression of ANK and/or ENPP and/or the activation, amount or activity of these proteins. The composition can be in the form of a cell or cells. The invention also relates to methods of using the composition.

The present disclosure provides compounds and pharmaceutically acceptable salt thereof, and methods of using the same. The compounds and methods have a range of utilities as therapeutics, diagnostics, and research tools. In particular, the subject compositions and methods are useful for reducing signaling output of oncogenic proteins.

This invention provides a method for stably producing airway epithelial cells from pluripotent stem cells. Specifically, the invention relates to a method for producing airway epithelial cells from pluripotent stem cells comprising steps: (1) culturing pluripotent stem cells in a medium containing activin A and a GSK3 inhibitor; (2) culturing the cells obtained in Step (1) in a medium containing a BMP inhibitor and a TGF inhibitor; (3) culturing the cells obtained in Step (2) in a medium containing BMP4, retinoic acid, and a GSK3 inhibitor; (5) subjecting the cells obtained after Step (3) to three-dimensional culture in a medium containing a GSK3 inhibitor, FGF10, and a ROCK inhibitor; and (6) subjecting the proximal airway epithelial progenitor cells obtained in Step (5) to three-dimensional culture in a medium containing a ROCK inhibitor.

The present invention relates to a method for obtaining an antibody from an avian B cell antibody library, comprising the following steps (a) to (d): (a) a step of allowing an avian B cell antibody library to come into contact with an antigen in the presence of a calcineurin inhibitor and avian serum, (b) a step of selecting avian B cells that bind to the antigen in the step (a), (c) a step of culturing the avian B cells selected in the step (b) in the presence of a calcineurin inhibitor and avian serum, and (d) a step of obtaining the avian B cells obtained through the step (c) and/or an antibody expressed by the avian B cells.