One aspect of the invention provides a method of generating three-dimensional biological structures. The method includes: (a) depositing a first layer of a suspension over a substrate, the suspension including a liquid and a plurality of cells; (b) allowing the plurality of cells to attach to the substrate and form a first layer of attached cells; (c) depositing a cell-attachment agent over the first layer of attached cells; and (d) depositing a second layer of the suspension over the cell-attachment agent.

A three-dimensional cell growth containment article is described, which includes a molded body channelized by removal of sacrificial channelizing element(s) therefrom, so that the molded body contains one or more channel(s) therein, with a matrix material in at least one of such channel(s) that is supportive of three-dimensional cell growth in the matrix material. A method for making such articles is also described, in which a molded body is formed with one or more sacrificial channelizing element(s) therein, following which the sacrificial channelizing element(s) are removed. The three-dimensional cell growth containment articles of the present disclosure may be utilized in any applications in which there exists a need to reproducibly generate three-dimensional cellular structures, e.g., islet transplantation for diabetes treatment, transplantation of hormone secreting cells, cellular scaffolds for wound healing, and generation of tissue engineering structures to regain structural usefulness for orthopedic applications.

Provided are a composition and a sheet, including a mesenchymal stem cells-hydrogel-biodegradable support or a mesenchymal stem cells-hydrogel-nondegradable support and a preparing method thereof. More specifically, in the sheet including a mesenchymal stem cells-hydrogel-biodegradable support or a mesenchymal stem cells-hydrogel-nondegradable support according to the present invention, the high-active mesenchymal stem cells may be applied to a wounded part of a patient with epidermolysis bullosa as it is without isolation using proteases, and in the culturing, an extracellular matrix such as collagen, laminin, fibronectin, and elastin secreted from the mesenchymal stem cells is wholly present on the hydrogel to have an advantageous effect that skin reproduction and re-epithelization abilities are significantly excellent as compared with conventional dressing agents used for epidermolysis bullosa.

Kits for making a spheroid-stabilizing hydrogel in a calcium-free or calcium-chelated cell culture media include (a) a gelation agent including a polygalacturonic acid (PGA) compound or an alginic acid compound, wherein the PGA compound includes at least one of: (i) pectic acid or salts thereof, or (ii) partially esterified pectic acid having a degree of esterification from about 1 to about 40 mol % or salts thereof; (b) a crosslinking agent, wherein the crosslinking agent includes a salt of a divalent ion; and (c) a proton donor, wherein the proton donor includes lactones, esters, or other compounds that hydrolyze in aqueous solutions to form acids over a period of from 10 minutes to 1 hour. Resultant spheroid-stabilizing hydrogels and methods of preparing the same.

The present disclosure relates to a 3D bioprinter (1) comprising a base unit (2). The base unit (2) has a support (3) adapted for mounting of at least one toolhead (4), a communication interface part (5) for communication of data with the at least one toolhead (4), when mounted, and a base unit processing element (7) adapted to communicate with a toolhead processing element (8) of the at least one toolhead over said communication interface part (5). The present disclosure relates further to a 3D bioprinter toolhead. The present disclosure relates further to a method for bioprinting a construct.

The present disclosure provides methods of lineage specific differentiation of pluripotent stem cells, including induced pluripotent stem cells, into floor plate midbrain progenitor cells, determined dopamine (DA) neuron progenitor cells, and/or DA neurons. Also provided are compositions uses thereof, such as for treating neurodegenerative diseases and conditions, including Parkinson's disease.

The present invention provides a scaffold for culturing retinal tissue comprising an amount of gelatin, an amount of chondroitin sulfate, an amount of hyaluronic acid, wherein the amount of gelatin, chondroitin sulfate, and hyaluronic acid are prepared into a three-dimensional monolith, wherein the monolith is sectioned into planar sheets, and an amount of laminin-521.

The invention provides a construct (1) comprising a number N of material types (100, 110, . . . ), wherein N is at least 2, wherein at least two of the material types (100, 110, . . . ) comprise granular material (101) comprising particles (10), wherein the granular material (101) at least defines an exterior surface (6) of the construct (1), wherein the construct (1) is self-supporting, and wherein the construct (1) is (i) self-healing or is (ii) configured for being self-healing by changing a liquid (15) content of the construct (1); wherein the different material types (100, 110, . . . ) mutually differ in at least one characteristic (19) selected from the group consisting of a physical characteristic and a chemical characteristic.

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.

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.