The present invention provides an improved method of producing highly pure retinal pigment epithelial (RPE) cells by differentiation of pluripotent stem cells.

[Problem] To provide a supporting bath useful for three-dimensional tissue culture and a method for producing cultured three-dimensional tissue by using the supporting bath. [Solution] Use of a supporting bath for three-dimensional tissue culture, that comprises polymer and water, wherein a thixotropic gel is formed in the bath with the gel being dissolvable in solvent. Also, use of a method for producing the supporting bath.

The present disclosure addresses IBD from the standpoint of inhibiting or ablating pathogenic mucosal stem cells cloned from defined regions of disease in the gastrointestinal tract. In the case of Crohn's disease, for example, isolation of those stem cells according to the methods of the present disclosure reveals a pattern of inflammatory gene expression in stem cells from the terminal ileum and colon that is epigenetically maintained despite months of continuous cultivation in the absence of immune or stromal cells, or of intestinal microbes. Superimposed on this distributed inflammatory phenotype is a differentiation defect that profoundly and specifically alters the mucosal barrier properties of the terminal ileum. The co-existence of diseased and normal stem cells within the same endoscopic biopsies of Crohn's disease patients implicates an epigenetically enforced heterogeneity among mucosal stem cells in the dynamics of this condition.

Embodiments of the disclosure encompass methods and compositions related to treatment of Acute Respiratory Distress Syndrome caused by any reason, including caused by a coronavirus, for example. In particular embodiments, fibroblasts are delivered to an individual in need thereof to stimulate generation of T regulatory cells that may or may not be FoxP3-positive, and/or immune regulatory cells previously exposed to fibroblasts are delivered to the individual.

Some embodiments are directed to a method for preparing a skin substitute, a dermal substitute, to a skin substitute, to a dermal substitute and to a kit for implementing the method. Some other embodiments are directed to a graft that can consist of of a skin substitute and to the use thereof as treating a skin disorder and/or a loss of skin substance.

Embodiments described herein relate generally to a three-dimensional ex vivo skeletal muscle tissue comprising a hydrogel and a plurality of cells that includes skeletal muscle cells, at least a portion of the cells being encapsulated inside the hydrogel. In some embodiments, the skeletal muscle tissue is characterized by one or more contractions in response to an electrical and/or chemical stimulation.

Disclosed are bioprinted, three-dimensional, biological skin tissues comprising: a dermal layer comprising dermal fibroblasts; and an epidermal layer comprising keratinocytes, the epidermal layer in contact with the dermal layer to form the three-dimensional, engineered, biological skin tissue. Also disclosed are arrays of engineered skin tissues and methods of making engineered skin tissues.

A method for producing a three-dimensional cell structure having a vascular network, including preparing a mixture of a cationic substance, a polyelectrolyte, an extracellular matrix component, and a cell population including endothelial cells, collecting, from the mixture, a cell aggregate including the cell population, the cationic substance, the polyelectrolyte, and the extracellular matrix component, and culturing a collected cell aggregate in a medium. The mixture includes the extracellular matrix at a concentration of 1.0×10.sup.−8 mg/mL or more and less than 2.5×10.sup.−2 mg/mL.

A method of producing a conditioned medium for culturing a patient-derived cancer cell, including culturing, in a first medium for 24 hours or longer, a three-dimensional cell tissue including an extracellular matrix component, a polymer electrolyte, and a cell cluster including a fibroblast, and collecting the first medium in which the three-dimensional cell tissue has been cultured as the conditioned medium.

Provided are live, artificial, skin substitute products and methods of making and using the same, such as for wound treatment and compound testing, including compound testing for efficacy, toxicity, penetration, irritation and/or metabolism testing of drug candidates or compositions such as cosmetics. Described herein is an artificial mammalian skin substitute product, comprising: (a) optionally, but in some embodiments preferably, a first (“hypodermis-like”) layer comprising live mammalian adipocytes (e.g., induced pre-adipocytes) in a first hydrogel carrier; (b) a second (“dermis-like”) layer contacting or directly contacting the first layer and comprising live mammalian fibroblast cells and' live mammalian follicle dermal papilla cells in combination in a second hydrogel carrier; (c) a third (“epidermis-like”) layer contacting or directly contacting the second layer (i.e., on the opposite side thereof as the first layer, so that the second layer is sandwiched between the first and third layers when the first layer is present), the third layer comprising live mammalian keratinocytes and live mammalian melanocytes in combination in a third hydrogel carrier.