The disclosure generally relates to methods and compositions for preparing a neural tissue construct. In particular, provided herein are methods for generating a neural tissue construct using glutamatergic cortical progenitor cells; GABAergic interneuron progenitor cells; and bio-ink.

Multiplanar microfluidic devices are provided that facilitate direct transverse fluid communication between a first microfluidic channel a plurality of adjacent microfluidic channels, where the adjacent microfluidic channels reside both laterally adjacent and vertically adjacent to the first microfluidic channel, thereby facilitating transverse diffusion to or from the adjacent microfluidic channels in both lateral and vertical directions. Geometrical meniscus-pinning features, such as meniscus-pinning ridge structures, are provided between adjacent microfluidic channels to restrict transverse flow between the microfluidic channels. Accordingly, a gel structure may be formed within the first microfluidic channel and one or more of the adjacent microfluidic channels can function as a perfusion channel, for example, for delivering media to cells residing withing the gel structure. Such devices may be extended and/or arrayed to include multiple channels with laterally and vertically adjacent perfusion microfluidic channels, optionally with shared lateral perfusion microfluidic channels among adjacent pairs of devices.

Provided is a cell culture substrate for trait induction control of a macrophage, which has a pattern of unevenness on a surface to which a cell adheres, the width of the unevenness being 50 nm or more and less than 1,000 nm.

A trait induction method of undifferentiated cells is provided, including: culturing undifferentiated cells on abase material which has an uneven pattern on the surface to which the cells adhere and of which the width of the unevenness is 1 nm to 1,000 nm.

A cell culture carrier includes a first substrate. The first substrate has a plurality of wells on an upper surface and includes a porous body. The porous body has pores with an average pore diameter of 0.05 μm or more and 10 μm or less, and has a porosity of 25% or more and 50% or less. The first substrate has a thickness from a lowermost portion of the well to a lower surface of the substrate of 50 μm or more and 10 mm or less.

A method for micro-tissue encapsulation of cells includes coating a tissue scaffold stamp with an extracellular matrix compound; depositing the tissue scaffold stamp onto a thermoresponsive substrate; seeding the tissue scaffold stamp with a cell culture; incubating the cell culture on the tissue scaffold stamp at a temperature that is specified, wherein the cell culture forms a cell patch that is attached to the extracellular matrix compound; removing the thermoresponsive substrate by lowering the temperature; removing the tissue scaffold stamp from the cell patch to form a micro-tissue structure by dissolving the tissue scaffold stamp in a solvent; folding the micro-tissue structure by suspending the micro-tissue in the solvent to enable the cell patch to fold the micro-tissue structure; collecting the folded micro-tissue structure from the solvent; and administering the folded micro-tissue structure to an organism.

Sub-millimeter scale three-dimensional (3D) structures are disclosed with customizable chemical properties and/or functionality. The 3D structures are referred to as drop-carrier particles. The drop-carrier particles allow the selective association of one solution (i.e., a dispersed phased) with an interior portion of each of the drop-carrier particles, while a second non-miscible solution (i.e., a continuous phase) associates with an exterior portion of each of the drop-carrier particles due to the specific chemical and/or physical properties of the interior and exterior regions of the drop-carrier particles. The combined drop-carrier particle with the dispersed phase contained therein is referred to as a particle-drop. The selective association results in compartmentalization of the dispersed phase solution into sub-microliter-sized volumes contained in the drop-carrier particles. The compartmentalized volumes can be used for single-molecule assays as well as single-cell, and other single-entity assays.

A cell culture matrix for a perfusion-flow fixed-bed reactor is provided. The cell culture matrix includes a substrate having a porous sheet for adhering cells thereto. The sheet has a first side, a second side opposite the first side, a thickness separating the first side and the second side, and a plurality of openings formed in the substrate, arrayed in a regular pattern, and passing through the thickness of the substrate. The porous sheet is wound into a cylindrical shape having a plurality of wound layers, and the cell culture matrix does not include a spacer material or physical barrier between the plurality of wound layers of the substrate.

A method for making an at least double layered cell aggregate and/or an artificial blastocyst, and/or a further-developed blastoid termed blastoid, by forming a double layered cell aggregate from at least one trophoblast cell and at least one pluripotent and/or totipotent cell, and culturing the aggregate to obtain an artificial blastocyst having a trophectoderm-like tissue that surrounds a blastocoel and an inner cell mass-like tissue. The cell aggregate can be formed from toti- or pluripotent stem cell types, or induced pluripotent stem cell types, in combination with trophoblast stem cells. Formation of a blastoid can be achieved by culturing the cell aggregate in a medium preferably comprising one or more of a Rho/ROCK inhibitor, a Wnt pathway modulator, a PKA pathway modulator, a PKC pathway modulator, a MAPK pathway modulator, a STAT pathway modulator, an Akt pathway modulator, a Tgf pathway modulator and a Hippo pathway modulator. Also, a method for growing an at least double layered cell aggregate into an artificial blastocyst, and into a further-developed blastoid, a foetus or a live animal and an in vitro cell culture comprising the mentioned compounds and/or cell aggregates.

Disclosed are a therapeutic agent for muscular dystrophy employing pluripotent stem cells obtained from dental pulp and a method for preparation thereof. The therapeutic agent for muscular dystrophy comprises pluripotent stem cell-enriched human dental pulp-derived cells as the active ingredient, and is prepared by a method of preparation comprising the steps of: (a) adding dental pulp-derived cells contained in a dental pulp suspension, in a culture vessel containing feeder cells whose proliferative ability is suppressed, onto a membrane having micropores that can block feeder cells from passing therethrough and supported within the vessel in a manner that avoids direct contact of the lower side face thereof with the feeder cells, and culturing the dental pulp-derived cells on the membrane while preventing direct contact with the feeder cells, and (b) recovering the cells having grown on the membrane as the pluripotent stem cell-enriched human dental pulp-derived cells.