In certain example embodiments, the invention provides a method of generating an ex vivo cell-based system comprising dissociating an original tissue sample obtained from a subject into a single cell population; determining an in vivo phenotype of the tissue sample by conducting single-cell RNA analysis on a first portion of the single cells; establishing an ex vivo cell-based system from a second portion of the single cells; and culturing the ex vivo cell-based system in a medium or conditions selected to maintain the in vivo phenotype. In some embodiments, the original tissue sample is a tumor tissue sample, such as a pancreatic ductal adenocarcinoma (PDAC) tumor sample.

An array platform for three-dimensional cell culturing and drug testing and screening is disclosed. In the array platform, a hydrogel-cell mixture injection area is configured to inject a plurality of kinds of hydrogel-cell mixtures. Cell observation areas are connected to the hydrogel-cell mixture injection area. Electrodes are disposed under the cell observation areas and automatic cell quantification and three-dimensional cell co-arrangement of the plurality of kinds of hydrogel-cell mixtures in the cell observation areas through the electrodes to imitate a structure of body's tissues. A drug injection area is configured to inject a plurality of kinds of drugs. Drug combination generators respectively correspond to the cell observation areas and are connected to the drug injection area. Each drug combination generator has a microfluidic channel structure and configured to generate drug combinations according to the plurality of kinds of drugs.

A developmental toxicity screening assay using spatially organized cardiac organoids with contracting cardiomyocytes in the center surrounded by stromal cells distributed along the pattern perimeter engineered from human induced pluripotent stem cells (hiPSCs). Cardiac organoids generated from 600 μm-diameter circles were used as a developmental toxicity screening assay for the quantification of the embryotoxic potential of nine pharmaceutical compounds. The cardiac organoids were demonstrated as having a potential use as an in vitro platform for studying organoid structure-function relationships, developmental processes, and drug-induced cardiac developmental toxicity.

A method of efficiently recovering a large amount of exosomes from mesenchymal stem cells is provided. The method includes: a three dimensional culture step of three dimensionally culturing mesenchymal stem cells in a medium containing sugar by using a nonwoven fabric as a scaffold; a post-plateau culture step of further culturing for a certain period of time after the amount of the sugar consumed by the mesenchymal stem cells reaches a plateau; and an exosome recovery step of recovering exosomes from the mesenchymal stem cells. The mesenchymal stem cells are adipose-derived mesenchymal stem cells.

A recombinant spider silk protein, consisting of no more than 800 amino acids, comprising a set of domains arranged according to the formula (NT)-REP-CT, wherein: the optional NT-domain, if present, comprises a sequence of 100 to 160 amino-acid residues derived from the N-terminal domain of a spider silk protein; the REP-domain comprises a sequence of 30 to 600 amino acid residues derived from the repetitive segment of a spider silk protein; and the CT-domain comprises a sequence of 70 to 120 amino acid residues derived from the C-terminal domain of a spider silk protein selected from: a sequence of 72 to 110 amino acid residues derived from the C-terminal domain of a spider silk protein, wherein the sequence comprises at least 7 residues independently selected from K, R, E and D; a sequence having at least 85% identity to SEQ ID NO: 15 or any one of SEQ ID NOs: 62-65 or 67-73; and a sequence having at least 70% identity to SEQ ID NOs: 64 or any one of SEQ ID NOs: 62-65 or 67-73, wherein the sequence comprises at least 7 residues independently selected from K, R, E and D.

There is provided an in vitro three dimensional cell construct for use as a model of the avian intestine derived from avian intestinal tissue comprising avian cells organised into intestinal villi and crypts. Suitably the construct comprises an exterior surface that mimics the apical surface of a chicken intestine. Also provided are methods of making the cell construct and use of the construct as an in vitro intestinal model system to examine an agent including, but not limited to a microbe, a vaccine, a pharmaceutical, a feed additive, a toxin, a pre-biotic, post-biotic, pre pro post biotic, therapeutic, a cell, gene construct, protein, immune-modulator, an intestinal effector agent, a candidate intestinal effector agent, cell signalling inhibitor, or cell signalling activator.

The present invention relates to a method for obtaining a neural micros-phere, comprising the steps of culturing pluripotent stem cells (PSCs), differentiating the PSCs into neural stem precursor blast cells, aggregating the neural stem precursor blast cells to form a neural microsphere, allowing the neural stem precursor blast cells of the neural microsphere to further mature, and collecting the neural microsphere.

The invention provides an isolated dehydrated corneal tissue, comprising a full thickness corneal stroma, and substantially all, or all, of the Bowman's membrane, wherein the stroma contains cellular material. Also provided is a method to product the tissue and uses of the tissue.

The present invention provides a pharmaceutical composition for the treatment of patients having ovarian cancer, lung cancer or mesothelioma and showing a Selection Factor of −30% or below, comprising a therapeutically effective amount of ipilimumab, and optionally a pharmaceutically acceptable diluent or carrier, wherein the patient is selected on the basis of a positive response to an ex vivo three-dimensional (3D) patient derived tumour culture, the method comprising: (a) preparing a three-dimensional, optionally size-normalised, tumour culture from a patient-derived tumour sample in a multitude of replicates; (b) adding one or more immunotherapeutic agents to the culture, and (c) culturing for a pre-defined time period; and (d) determining the effect that the one or more immunotherapeutic agents has on the tumour cell aggregates by measuring the total area of objects in the culture that are above a threshold area associated with tumour cell aggregates, and the total area of objects that are below a threshold associated with immune cells, using three-dimensional imaging of the cell culture; wherein if following culturing with a composition comprising ipilimumab, the total area of the large objects decreases and/or the total area of the small objects increases relative to a control the patient is treated with ipilimumab.

Engineered tissue models based on three-dimensional (3D) scaffolds, also referred to herein as tissue-engineered 3D models, can be used as in vitro diagnostic and drug screening tools for predicting, preventing and/or treating cancer metastases.