A 3D cell culture gel kit and a 3D cell culture method using the same are provided. The 3D cell culture gel kit includes a gel material A, a buffer solution C, and a buffer solution D. The 3D cell culture method includes the steps of adding cells into a mixed solution containing the gel material A and setting the mixed solution at low temperature to get gel containing the cells. Then adding the buffer solution C to the gel for performing crosslinking. Next removing the buffer solution C and adding a growth medium. Let stand until the cells form spheroids in the gel. Moreover, the buffer solution D is used to dissolve the gel and the cells cultured are taken out for analysis. Thereby the 3D cell culture gel kit is convenient to use and suitable for 3D culture of a plurality of cell lines.

A method for producing a medium composition for suspension culture of an adherent cell, including the following steps: (i) a step of making an extracellular matrix carried on a nanofiber composed of water-insoluble polysaccharides, (ii) a step of adding the extracellular matrix-carrying nanofiber obtained in step (i) to a medium is provided by the present invention.

The present invention is directed to a system including neurons over-expressing UBB+1, organized in a 3-dimensional culture, and method of using same. A process for making the system of the invention is also provided.

Disclosed are methods for making a vascularized hollow organ derived from human intestinal organoid (HIOs). The HIOs may be obtained from human embryonic stem cells (ESC's) and/or induced pluripotent stem cells (iPSCs), such that the HIO forms mature intestinal tissue. Also disclosed are methods for making a human intestinal tissue containing a functional enteric nervous system (ENS).

The present invention relates to a chemically defined medium for eukaryotic cell culture, comprising water, at least one carbon source, one or more vitamins, one or more salts, one or more growth factors, one or more fatty acids, one or more buffer components, selenium and one or more further trace elements and its use in the culture of cancer stem cells, in particular tumorsphere culture of cancer stem cells.

Systems, apparatus and methods for producing objects with cryogenic 3D printing with controllable micro and macrostructure with potential applications in tissue engineering, drug delivery, and the food industry. The technology can produce complex structures with controlled morphology when the printed 3D object is immersed in a liquid coolant, whose upper surface is maintained at the same level as the highest deposited layer of the object. This ensures that the computer-controlled process of freezing is controlled precisely and already printed frozen layers remain at a constant temperature. The technology controls the temperature, flow rate and volume of the printed fluid emitted by the dispenser that has X-Y positional translation and conditions at the interface between the dispenser and coolant surface. The technology can also control the temperature of the pool of liquid coolant and the vertical position of the printing surface and pool of coolant liquid.

A cell culture apparatus is provided, including a storage tank including one or a plurality of cell culture units, in which the cell culture unit includes a culture chamber having an inner surface-side space in which a culture solution is stored, a permeable membrane having a first surface to which cells are adherable and a second surface opposite to the first surface, the first surface facing the inner surface-side space, a culture solution storage chamber that stores the culture solution, a culture solution introduction flow path that introduces the culture solution in the culture solution storage chamber to the inner surface-side space, and a culture solution discharge flow path that sends, to the culture solution storage chamber, the culture solution which permeates through the membrane from the inner surface-side space and flows into an outer surface-side space that the second surface of the membrane faces.

The present invention relates to a method for in-vitro production of mammalian neurons expressing the 6 isoforms of the Tau protein (2N4R, 1N4R, 0N4R, 2N3R, 1N3R, 0N3R), comprising a step of neuronal differentiation, in which cellular microcompartments are cultivated for a period of 5 weeks to 100 weeks, each one comprising a hollow hydro gel capsule surrounding post-mitotic neuronal cells and an extracellular matrix, the neuronal differentiation step being carried out in a bioreactor, the cellular microcompartments being kept in suspension in an enclosure of the bioreactor containing a neuronal differentiation medium.

The instant disclosure is directed to a method for vascularizing a pancreatic islet comprising culturing the pancreatic islet or β-cells with an endothelial cell comprising an exogenous nucleic acid encoding an ETV2 transcription factor under conditions wherein the endothelial cell expresses the ETV2 transcription factor. The instant disclosure is further directed to a method for making a vascularized β-cell organoid comprising culturing the pancreatic islet or β-cells with an endothelial cell comprising an exogenous nucleic acid encoding an ETV2 transcription factor under conditions wherein the endothelial cell expresses the ETV2 transcription factor. Disclosed also are vascularized islets and vascularized β-cell organoids produced by the methods of the instant disclosure, as well as methods for using the same.

A production method for a cerebral organoid having amyloid plaques is provided, the method including a step (a) of forming, in the presence of a SMAD inhibitor, an embryoid body from a pluripotent stem cell having a mutation in an Alzheimer's disease-related gene; a step (b) of embedding the embryoid body after the step (a) in an extracellular matrix and three-dimensionally culturing the embedded embryoid body in the presence of a SMAD inhibitor and a glycogen synthase kinase 3β (GSK3β) inhibitor to form an organoid; and a step (c) of removing the organoid after the step (b) from the extracellular matrix and subjecting the removed organoid to stirring culture in a medium, where at least a part of the step (c) is carried out in the presence of leukemia inhibitory factor (LIF).