Disclosed herein are methods, compositions, kits, and agents useful for inducing ? cell maturation, and isolated populations of SC-? cells for use in various applications, such as cell therapy.

The invention provides for methods of differentiating pancreatic endocrine cells into pancreatic beta cells expressing PDX1, NKX6.1, MAFA, UCN3 and SLC2A. These pancreatic beta cells may be obtained by step-wise differentiation of pluripotent stem cells. The pancreatic beta cells exhibit glucose-dependent mitochondrial respiration and glucose-stimulated insulin secretion similar to islet cells.

The present invention provides methods, cell cultures and differentiation media to promote differentiation of pluripotent stem cells to pancreatic endocrine cells of a mature phenotype. The resulting pancreatic endocrine cells express single hormonal insulin, PDX1, NKX6.1, and MAFA. In one or more differentiation stages, culturing may be carried out in a culture vessel at the air-liquid interface.

Disclosed herein are methods, compositions, kits, and agents useful for inducing ? cell maturation, and isolated populations of SC-? cells for use in various applications, such as cell therapy.

Compositions for generating a pancreatic beta-like cell population from a population of undifferentiated cells and methods of use thereof, are provided. The method is an 8 stage process interrupted by a priming step, and it includes; using a chemically defined protocol for the efficient generation of pancreatic progenitors (PPs); improved assembly PPs into 3D clusters; a priming step which uses a chemical/factor cocktail (PP-10C) to maintain 3D-PPs status and enhances their potential to differentiate into ? cells ;and a 3-step differentiation protocol using select chemical cocktails that efficiently converts PP-10C-treated 3D-PPs into functional ? cells.

The disclosed methods result in a population of functional ? cells which expresses pancreatic cell markers selected from the group of c-peptide, the transcription factors NKX6.1, PDX1, PAX6, NEUROD1 and are INS+, and which do not express substantial levels Glucagon (GCG).

The functional ? cells can be used to treat conditions such as diabetes.

Disclosed herein are methods, compositions, kits, and agents useful for inducing ? cell maturation, and isolated populations of SC-? cells for use in various applications, such as cell therapy.

The present invention discloses a method for culturing human limbal stem cells comprising the steps of: pre-treating a feeder cell; seeding the feeder cells in a porous membrane; placing the porous membrane in a container to separate the container into a first cell culture compartment and a second cell culture compartment, wherein the feeder cells are located in the second cell culture compartment; injecting the culture medium into the container; and placing the human limbal stem cells in the first cell culture compartment. Compared to the prior art, the method for culturing human limbal stem cells of the present invention utilizes the porous membrane to culture the human limbal stem cells to make the expansion rate better than the traditional culture method. In addition, the culture medium used in the present invention does not contain the dimethyl sulfoxide (DMSO) with cytotoxicity to reduce the probability of cell death.

Disclosed herein are methods, compositions, kits, and agents useful for inducing ? cell maturation, and isolated populations of SC-? cells for use in various applications, such as cell therapy.

Disclosed herein are methods, compositions, kits, and agents useful for inducing ? cell maturation, and isolated populations of SC-? cells for use in various applications, such as cell therapy.

Disclosed herein are methods, compositions, kits, and agents useful for inducing ? cell maturation, and isolated populations of SC-? cells for use in various applications, such as cell therapy.