A method for isolating oogonial stem cells (OSCs) including forming an ovarian cell suspension from an ovary, forming ovaroids by culturing the ovarian cell suspension in a three-dimensional culture, and migrating the OSCs around the ovaroids by culturing the ovaroids on a mouse embryonic fibroblast (MEF)-coated plate.

The present disclosure provides a method of deriving and maintaining a midbrain-like organoid in culture, comprising (a) culturing pluripotent stem cells to obtain neuronal lineage embryoid bodies; (b) culturing the neuronal lineage embryoid bodies from (a) to obtain midbrain regionalized tissues; (c) embedding and culturing the midbrain regionalized tissues from (b) in an extracellular matrix to obtain neuroepithelial tissues; and (c) culturing the neuroepithelial tissues from (c) to obtain a midbrain-like organoid. Also disclosed herein are culture media suitable for deriving and maintaining neuronal lineage embryoid bodies comprising (a) TGF- Inhibitor and/or SMAD2/3 inhibitors; and (b) WNT-signaling activator; culture media suitable for deriving and maintaining midbrain regionalized tissues comprising (a) TGF- Inhibitor and/or SMAD2/3 inhibitors; (b) WNT-signaling activator; (c) hedgehog signaling protein; and (d) fibroblast growth factor; culture media suitable for deriving and maintaining neuroepithelial tissues comprising (a) hedgehog signaling protein; and (b) fibroblast growth factor and culture media suitable for deriving and maintaining a midbrain-like organoid comprising (a) neurotrophin factor; (b) ascorbic acid; and (c) activator of cAMP-dependent pathway.

A spheroid tissue microarray comprises an array of tissue spheroids embedded within a porous mold. The product may be impregnated with a wax or resin and sectioned, and contains spheroids which are precisely located in a regular geometric grid. A method of manufacturing a spheroid tissue microarray comprises the steps of: forming a mold of porous material from liquid mold material in a casting mold, and allowing the liquid mold material to set; removing the porous mold from the casting mold; topping up the porous mold with further liquid mold material, and allowing recesses to form in the surface of the mold by the drawing-in of liquid mold material through shrinkage as the liquid mold material sets; placing tissue spheroids into the recesses in the surface of the porous mold; and sealing the tissue spheroids within the mold by topping off with liquid mold material and allowing the liquid mold material to set. An alternative method comprises the steps of: forming a mold of porous material from liquid mold material in a casting mold; allowing the liquid mold material to set; removing the porous mold from the casting mold; placing spheroids in recesses at the bases of wells in the mold of porous material; and sealing the spheroids within the porous mold by adding further porous material on top of the spheroids; wherein the recesses at the bases of the wells in the porous material are formed by protrusions of the casting mold carrying further, nipple-shaped, protrusions.

The invention provides a callus induction medium for blueberry tissue culture, taking woody plant medium (WPM) as a basic medium, and including: 0.5-5.0 mg/L forchlorfenuron (CPPU) and 0.1-0.4 mg/L 2-isopentenyladenine (2-ip). The present invention also provides a callus culture method for blueberry, including inoculating the blueberry explant into the above callus induction medium to conduct induction culture in order to form blueberry callus. The present invention also discloses the medium combination and culture method to culture the above blueberry callus to blueberry tissue culture plant. For the above medium and culture method, the differentiation effect is good, efficiency is high, one can conduct continuous differentiation, and the effect is better on multiple varieties.

The present invention provides destructible, digestible, or dissolvable microwell arrays, such as hydrogel microwell arrays, which are useful for segregating, culturing, and analyzing biological samples such as cells and mixtures of cells.

Disclosed is a method for manufacturing a composite nanofiber support and a fish collagen/synthetic polymer nanofiber support manufactured by the method, wherein the method comprises: a first step for dissolving a synthetic polymer in an organic solvent; a second step for dissolving a fish collagen in water to prepare an aqueous fish collagen solution; a third step for adding the aqueous fish collagen solution prepared in the second step to the synthetic polymer solution prepared in the first step, followed by mixing; and a fourth step for electrospinning the mixture solution prepared in the third step to manufacture a nanofiber support.

The present invention relates to a process for producing a droplet assembly, which droplet assembly comprises a plurality of droplets, wherein each of said droplets comprises: an aqueous medium comprising a hydrogel compound; and one or more biological cells disposed in the aqueous medium, which process comprises: generating, in a bulk hydrophobic medium, a plurality of droplets, wherein each of said droplets comprises: an aqueous medium comprising a hydrogel compound; and one or more biological cells disposed in the aqueous medium. The invention also relates to a droplet assembly comprising a plurality of droplets, wherein each of said droplets comprises: (i) an aqueous medium comprising a hydrogel compound; (ii) one or more biological cells disposed in the aqueous medium; and (iii) an outer layer of amphipathic molecules around the surface of the aqueous medium, wherein at least one droplet in the droplet assembly contacts at least one other droplet in the droplet assembly forming a layer of amphipathic molecules as an interface between contacting droplets.

The present disclosure relates to a cryopreserved pharmaceutical composition comprising immature dental pulp stem cells (IDPSCs) expressing CD13 and CD44 and methods of treating a neurological disease or condition comprising systemically administering to a subject a cryopreserved pharmaceutical composition comprising IDPSCs expressing CD13 and CD44. In some aspects, the IDPSCs in the cryopreserved pharmaceutical composition described herein further expresses beta-3-tubulin. In some aspects, the IDPSCs secrete brain-derived neurotrophic factor.

Ex-vivo culture systems are provided. Accordingly there is provided a culture system comprising a culture medium and a precision-cut tissue slice placed on a tissue culture insert, wherein the precision-cut tissue slice is maintained in a highly oxygenated atmosphere containing at least 50% oxygen and wherein said culture is rotationally agitated facilitating intermittent submersion of the tissue slice in the culture medium. Also provided are methods of culturing a tissue and methods of using the culture system for selecting a drug for the treatment of a disease.

The present invention relates to compositions comprising a polymeric matrix or a gel containing functional enzymes capable of re-creating under culture conditions the cell microenvironment existing in vivo. The present invention also relates to devices for cell cultures comprising such compositions, in particular hydrogel and the use thereof to control the chemical microenvironment of a cell culture or mimic physiological or pathological conditions of the in vivo cells. The compositions and the devices described herein could be also used in vitro for evaluating the therapeutic effect of a compound on a determined cell line or on primary cells.