Provided is a cell culture method comprising the step of culturing cells using a medium containing a laminin fragment having integrin binding activity, the method not comprising the step of coating a culture vessel with a laminin or a laminin fragment before seeding the cells in the culture vessel. The cell culture method of the present invention uses a smaller amount of a laminin fragment and still achieves a comparable culture efficiency as compared with the conventional cell culture method that uses a culture vessel precoated with a laminin or a laminin fragment.

The present invention relates to a method of expanding pluripotent stem cells (PSC) in suspension culture in a bioreactor, the method comprising (i) adding an inhibitor of ROCK (ROCKi) to pluripotent stem cells being cultivated in suspension in the bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of the pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding an excess volume of culture medium sufficient to decrease the concentration of the cell dissociation agent to a concentration at which cell aggregates can form again; and (iv) culturing of the mixture obtained in step (iii) under suitable conditions that allow the expansion of the PSCs.

High-yield mammalian transient expression systems can include a cell culture media (particularly serum free, non-animal derived, and/or chemically defined media) for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). Cells containing such introduced materials can then be cultured in the cell culture media. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly mammalian cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention also relates to compositions and kits useful for culturing and transforming/transfecting cells.

The present invention provides a new method for producing Engineered Heart Muscle (EHM) under chemically fully defined conditions all compatible with GMP regulations. The resulting human myocardium generates force and shows typical heart muscle properties.

Disclosed herein are novel methods for hydrolyzing eggshell membrane (ESM). In one embodiment, the method includes cultivating thermophilic bacteria in a solution containing 1-10% (wt %) ESM to decompose the ESM into the ESM hydrolysate; wherein, the thermophilic bacteria grow on the ESM as their sole source of nutrient. In another embodiment, the method includes treating ESM with a keratinase in the presence of a reducing agent at a condition sufficient to produce the ESM hydrolysate, in which the keratinase, the reducing agent, and the ESM are present in a weight ratio of 1:120:600. The thus produced ESM hydrolysate is enriched in essential amino acids, collagen, peptides and glycosaminoglycans.

The disclosure provides methods and compositions for affecting the development of antigen presenting cell (APC, e.g., a macrophage or dendritic cell). The methods include maturing an APC, promoting anti-inflammatory phenotype, promoting development of a T regulatory cell (Treg) from a naive T cell. The methods generally include exposing an APC to a tryptophan derived microbiota metabolite (TDMM), such as an anti-inflammatory or pro-mucosal TDMM, and permitting the APC to mature. In some embodiments, the conditioned APC is exposed to a naive T cell to further promote development of a T regulatory cell (Treg). In some embodiments, the TDMM is selected from the group consisting of indole, indole-3-acetate, 5-hydroxyindole, and indole-3-pyruvate.

The present invention discloses formulation of a serum-free medium used for human pluripotent stem cells, which comprises the following raw materials: inorganic salt components, organic components, amino acids and amino acid salts, energy substances and metabolic intermediates, vitamins and antioxidants, proteins and polypeptides, trace elements and chromogenic substances; while the culture process comprises the following steps: selecting a basic formulation, performing combination screening, identifying and evaluating results, and testing a new formulation of culture; and proportioning according to the following methods: adding aforesaid raw materials into 950 ml of water for injection, stirring gently until dissolved, and finally adding 2.438 g of sodium bicarbonate, and stirring gently until dissolved, and then adding 1 liter of water for injection, adjusting the pH to the desired value with 1 mol/L sodium hydroxide solution or 1 mol/L hydrochloric acid solution, finally filtering sterilized with 0.1 μm diameter filter under positive pressure, and storing the medium solution in dark place at 2° C.-8° C., the invention solves the problem of high cost of domestic import of serum-free formulation.

A method including adding choline at a specific concentration to a medium is useful for culturing cells.

The present invention relates to ex vivo methods for obtaining populations of human keratinocytes derived from human pluripotent stem cells.

A method for producing an epithelial cell sheet, comprising culturing cells derived from oral mucosal epithelial cells on a substrate in a serum-free medium, wherein the serum-free medium comprises (i) EGF protein or KGF protein, (ii) B-27 supplement, and (iii) a ROCK inhibitor.