178results about "Tissue screening" patented technology

The invention belongs to the technical field of tissue engineering, and particularly discloses a preparation method of a 3D full-thickness skin model. The preparation method provided by the invention comprises the following steps: constructing a corium layer, constructing an inner epidermal layer through first epidermal inoculation, constructing an epidermal layer through second epidermal inoculation and obtaining a complete 3D full-thickness skin model through co-culture. The 3D full-thickness skin model obtained through the preparation method is distinct in gradation, uniform in thickness, free of shrinkage and stable in quality, the epidermal layer and the corium layer are uniformly fused, and the 3D full-thickness skin model is suitable for skin function research and sample evaluation. Meanwhile, the preparation method provided by the invention is simple in process and convenient to operate, does not need additional reagents or anti-shrinkage tools, can meet the requirements of large-scale industrial production, and has good production feasibility and market application prospects.

A robust nasal organoid culture system and fully characterized 2D and 3D in vitro produced organoids are provided, as well as methods of using the organoids. The nasal organoids comprise a population of differentiated nasal epithelial cells having cellular heterogeneity and molecular markers similar to mature nasal epithelial tissue. Improved methods for producing two-dimensional and three-dimensional nasal organoids are also provided. These methods generally require a two-phase culture system, i.e., amplification and differentiation culture. The provided nasal organoids are suitable for reproducing nasal tissue complexity, for mimicking respiratory diseases, studying barrier function and drug delivery, high throughput drug screening, and assessing the effects of pathogens and environmental toxins on nasal respiratory epithelium.

The present invention relates to a method for preparing a mammalian vascular network or mature mammalian bone marrow organoid. Also provided is the use of the prepared vascular network or mammalian bone marrow organoid in the treatment of bone marrow-related diseases, in vitro generation of BMO or mammalian blood cells, as a model system for the pathogenesis of bone marrow-related diseases, and as a system for identifying and / or testing pharmaceutically effective compounds for the treatment or prevention of bone marrow-related diseases.

The invention provides a co-culture system of human alveolar epithelial cells and endothelial cells, which is used for researching interaction in an epithelial-endothelial barrier in lungs. It is indicated that infection with the highly pathogenic influenza A virus A / HK / 483 / 97 (H5N1) can significantly reduce alveolar fluid clearance (AFC) and increase protein permeability (APP). Administration of therapeutic extracellular vesicles (EV) after infection effectively restores AFC and APP, reduces viral replication, and enhances expression of antiviral genes. According to the invention, the treatment potential of EV is highlighted, and a valuable tool is provided for researching acute lung injury and improving a lung drug delivery technology.

The present invention relates to a damaged skin organoid model and a drug screening method using same, and is a technology that uses a technique for producing an air-liquid interface skin organoid, in which skin cells, appendages, nerve cells, adipocytes, and the like are formed as a structure similar to actual skin, and which produces damaged skin by irradiating the skin organoid with ultraviolet rays that simulate ultraviolet radiation reaching the earth's surface. In the present invention, by irradiation with simulated solar ultraviolet rays including 94.5% of UV-A and 5.5% of UV-B, the skin barrier was damaged and collagen degradation was promoted by increasing the activity of matrix metalloproteinases, and, at the same time, the characteristics of UV-damaged skin, such as increased secretion of pro-inflammatory cytokines, were successfully mimicked. Therefore, it is determined that the present invention will be effectively used as a UV-induced damage and photoaging model, and will serve as a useful technology for screening for potential therapeutic drugs.

The present disclosure provides methods of making tissue-derived epithelial organoids. In particular, the present disclosure provides a tissue-derived epithelial organoid embedded in a hydrogel suspended in a culture medium. The present disclosure further provides methods of using such organoids.

This method for culturing a cancer organoid includes culturing the cancer organoid on a top surface or inside of a cell structure in a medium not including an extracellular matrix, and the cell structure includes cells constituting stroma, and has two or more cell layers laminated in a thickness direction. A method for screening a test substance includes culturing a cell structure including the cancer organoid obtained by the method for culturing a cancer organoid in a presence of the test substance, and evaluating an effect of the test substance on the cancer organoid.

The present disclosure relates to methods and compositions for generating topographically organized tissues in vitro, for the resulting cultured tissue and components thereof, and for uses of such cultured tissue and its components in drug discovery, toxicology studies, and therapy.

A gut organoid includes a tissue structure that encloses a cavity. A method of using the gut organoid includes providing a test drug to the gut organoid in the presence of a buffer solution so that liquid comprising the test drug enters into the cavity, and collecting and analyzing the liquid in the cavity. A method for producing the gut organoid includes plating cells on a cell culture substrate including a surface on which a cell-adhesive region and a non-cell-adhesive region surrounding the cell-adhesive region are formed, and culturing the cells plated on the cell culture substrate so as to induce differentiation of the cells.

To provide an intestinal organoid equipped with the same function as an intestine, and a method for manufacturing the organoid.SOLUTION: An intestinal organoid of the present invention is derived from an embryonic stem cell and / or an artificial pluripotent stem cell, has a structure including a cavity, and a length in a long axis direction is 5 mm or more. A method for manufacturing an intestinal organoid of the present invention includes: a process 1 of inoculating a cell selected from an embryonic stem cell and an artificial pluripotent stem cell onto a substrate and a cell culture substrate including a cell adhesion region and a cell non-adhesion region surrounding the cell adhesion region which are formed on the surface of the substrate; and a process 2 of culturing a cell inoculated in the process 1. The process 2 includes differentiating a part of a cell into an entodermal cell and differentiating a part of a cell into an ectodermal cell.SELECTED DRAWING: Figure 1D

Provided are: an agent for organoid formation and proliferation in the absence of an extracellular matrix, the agent including a Hippo signaling pathway inhibitor as an active component; a method for organoid proliferation; an organoid proliferated by the above-described proliferation method; an agent for establishing an organoid in the absence of an extracellular matrix; a method for producing an organoid; an organoid produced by the above-described production method; a regenerative medicine preparation; and a method for screening for an agent that enables culture of an organoid in the absence of an extracellular matrix.

The present invention relates to purified biocompatible mucus capable of forming a gel. The mucus of the present invention may be provided in a tissue model along with a cell population. The mucin or tissue model of the present invention may be useful in in vitro models of digestion, mucus penetration, and / or epithelial transport.

This method for producing a brain organoid including aggregated tau protein comprises: a step (a) for culturing pluripotent stem cells in the presence of SMAD inhibitor to form embryoid bodies; a step (b) for embedding the embryoid bodies in an extracellular matrix and three-dimensionally culturing the embryoid bodies in the presence of SMAD inhibitor and GSK3β inhibitor to form an organoid that includes neural precursor cells; a step (c) for removing the organoid from the extracellular matrix and suspension-culturing the organoid in the presence of LIF to form a brain organoid; a step (d) for forcing the brain organoid to express a modified MAPT gene; and a step (e) for further suspension-culturing the brain organoid after step (d) to obtain a brain organoid having an aggregated tau protein.

Provided is a robust nasal organoid culture system and fully characterized 2D and 3D in-vitro generated organoids, and methods of using the organoids. The nasal organoids contain a population of differentiated nasal epithelial cells having cellular heterogeneity and molecular markers similar to mature nasal epithelial tissue. Improved methods for generating two-dimensional and three-dimensional nasal organoids are also provided. The methods generally require a two-phase culture system, i.e., expansion and differentiation culture. The provided nasal organoids are suitable for recapitulating nasal tissue complexity for modeling respiratory diseases, studying barrier function and drug delivery, high-throughput drug screening, and assessing the effects of pathogens and environmental toxins on the nasal respiratory epithelium.

What is provided is a nervous system cell-containing spheroid having suitable drug responsiveness. There are provided a nervous system cell-containing spheroid including at least one kind of nerve cells and nervous system cells other than the nerve cells, in which the proportion of the volume of one cell layer positioned at an outer edge of the spheroid to the volume of the entire spheroid is 15% to 30%; a method for producing the nervous system cell-containing spheroid; and a method for evaluating a test substance, the method including a step of incubating a nervous system cell-containing spheroid in the presence of a test substance, and a step of evaluating an effect of the test substance on the nervous system cell-containing spheroid, in which the nervous system cell-containing spheroid includes at least one kind of nerve cells and nervous system cells other than the nerve cells, and the proportion of the volume of one cell layer positioned at an outer edge of the spheroid to the volume of the entire spheroid is 15% to 30%.

The present invention relates to a method for producing a dermal or skin or epidermal equivalent on a two-layer substrate comprising a layer of at least 200 nm in thickness and having a porosity less than or equal to 5 μm formed by electrospinning of a composition comprising at least one polymer, and a layer of at least 20 μm in thickness and having a porosity greater than or equal to 20 μm formed by electrowriting of a composition comprising at least one polymer. The application also relates to the skin or dermal or epidermal equivalent that can be obtained with said method, the use of a dermal equivalent or a skin equivalent or an epidermal equivalent for screening compounds and finally the use thereof in wound dressing or for skin grafts.

The present invention relates to a purified, biocompatible mucus, which has the ability to form a gel. The mucus of the invention may be provided in a tissue model, with a cell population. The mucin or tissue model of the invention may be useful in an in vitro model of digestion, mucus permeation and / or epithelial transport.

To provide, in one embodiment, a method of using human-induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery.SOLUTION: In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.SELECTED DRAWING: None

The field of the invention is in vitro assays for the production of in vitro organ culture systems and the use of such in vitro organ culture systems for screening assays in drug screening, patient selection and personalized medical context. More specifically, the present invention relates to an in vitro method of establishing a tumor organoid from a cancer tissue sample obtained from a subject, an in vitro method of providing immune cells activated by an autologous tumor organoid, a method of preparing an in vitro organ culture system for mimicking interaction of a tumor with an immune system of a subject, and a method of preparing an in vitro organ culture system for mimicking interaction of a tumor with an immune system of a subject. The invention relates to an in vitro organ culture system, and to an in vitro organ culture system produced thereby, to a method for determining the reactivity of a tumor to at least one immunooncology drug therapy using said in vitro organ culture system, and to a kit for preparing an in vitro organ culture system that mimics the interaction of a tumor with the immune system of a subject, as disclosed herein.

The present disclosure provides a triple co-culture of a patient-derived cancer cell (PDC), a non-tumor derived cell that supports growth of the PDC and, optionally, may serve as an internal normal cell control (a "Tox Control" or "TC" cell), and a tumor-derived cell that is not derived from the patient and that exhibits sensitivity to at least one anti-cancer drug or drug combination (a "System Control" cell). The present disclosure also provides methods of use such triple co-culture systems, in for example, ex vivo personalized medicine, drug-discovery, drug development, and pre-clinical validation of candidates for clinical trials.

And a means for further improving the evaluation precision is provided in the aspect of evaluating pharmacokinetics / toxicity and efficacy by using the cell aggregate of a plurality of samples. A plate having a plurality of holes in which a liquid and an aggregate of kidney cells are accommodated, the openings of the holes being covered with a film, and the ratio of the total volume of the aggregate and the liquid relative to the total volume of the holes being a predetermined ratio.

The present invention provides a method for constructing a vascularized organoid spheroid model, a vascularized organoid spheroid model, and a use thereof. The construction method comprises the following steps: S100: preparing or acquiring an organoid, subjecting the organoid to digestion to obtain organoid cells, and acquiring fibroblasts and vascular endothelial cells; S200: mixing the organoid cells, the fibroblasts and the vascular endothelial cells, and resuspending with a culture medium, to obtain an organoid multicellular suspension; S300: inoculating the organoid multicellular suspension into a culture chamber having a U-shaped bottom, or a culture well having a U-shaped bottom, performing spheroid culture, and self-assembling, to form a vascularized organoid spheroid model. The present disclosure uses co-culture of organoid cells, fibroblasts and vascular endothelial cells, and also uses a low-adhesion U-shaped well plate to form microspheres, so as to prepare the vascularized organoid spheroid aggregate model.

The present invention relates to an intestinal epithelium model faithfully reproducing the pathophysiological mechanisms observed in vivo in the context of an inflammatory state of the intestinal barrier. The model according to the present invention comprises two compartments separated by a semi-permeable membrane. The first compartment, corresponding to the apical pole of the intestinal epithelium, comprises a coculture of Caco-2 cells differentiated into enterocytes and HT29-MTX cells differentiated into goblet cells. The second compartment, corresponding to the basolateral pole of the intestinal epithelium, comprises a culture of THP-1 monocytic cells differentiated into macrophages.The model according to the present invention is characterized in that the cells contained in the first compartment produce interleukin (IL-6) at a concentration greater than 100 pg / mL, and interleukin 8 (IL-8) at a concentration greater than 150 pg / mL, and the cells contained in the second compartment produce tumor necrosis factor α (TNF-α) at ​​a concentration greater than 40 pg / mL and interleukin 1β (IL-1β) at a concentration greater than 90 pg / mL. The present invention also relates to the method for obtaining this model as well as to a method for selecting candidate compounds for the treatment of inflammatory bowel diseases by verifying their ability to stop inflammation upon increase in permeability or to verify the restoration of permeability and the mucus layer.

This invention relates to the field of microfluidic chip technology, specifically to a dynamic microfluidic chip device, its preparation method, and its applications for studying CAR-T therapy for gliomas. The preparation method includes: fabrication of the microfluidic chip; reconstruction of neural tissue; reconstruction of the blood-brain barrier; and simulation of the glioma microenvironment. The dynamic microfluidic chip device includes a microfluidic chip; the microfluidic chip includes, from left to right, a vascular channel, a brain parenchyma channel, a tumor channel, and a subarachnoid channel; each of the vascular channel, brain parenchyma channel, tumor channel, and subarachnoid channel includes a cell culture chamber with an inlet on one side; the cell culture chambers of the vascular channel and the subarachnoid channel have corresponding outlets on the other side. The applications include: CAR-T cell infusion; observation of transmembrane migration; and detection of biomarkers. This invention, through the dynamic microfluidic chip device, its preparation method, and its applications, aims to achieve more accurate in vitro evaluation of CAR-T cell therapy.

The present invention discloses, in one embodiment, a method of using human induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery⋅. In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.

The present disclosure is directed to methods of producing a predictive model or human avatars for assessing Central Nervous System (CNS) disorders using organoids, optionally with artificial intelligence (Al).

A method for culturing a cancerous organ, which comprises a step for culturing a cancerous organ on the top surface or inside of a cell structure in which two or more cell layers are laminated in the thickness direction, said cell structure containing cells constituting a mesenchymal substance, with a culture medium that does not contain an extracellular matrix. A method for screening a substance to be tested, the method comprising: a step for culturing a cell structure comprising a cancerous organ obtained by the method for culturing a cancerous organ in the presence of a substance to be tested; and a step for evaluating the influence of the substance to be tested on the cancerous organ.

Provided is a structure composed of a cell population comprising endothelial cells, astrocytes and pericytes, and a 3D (three dimensional) cell growth material within which the cell population is located. The structure has a TEER value of at least 450 Ω / cm2. The cells of the structure may be derived from the brain. The cells may be human cells, and in particular may be primary derived non-immortalised cells. The structure is particularly suited for use in a model of the blood brain barrier, and the invention also provides such a model. The structure is located in a container, in which it separates a first chamber located on a first side of the structure and a second chamber located on a second side of the structure. The first and second chambers respectively contain first and second liquids in contact with first and second sides of the structure. The liquids mimic the brain extracellular fluid and the blood. The blood brain barrier model provided may be used in models of brain disease, and to investigate uptake of agents into the brain or diseased brain.

The present invention relates to a three-dimensional organotypic skin model of hydrogel comprising: - an epidermis of human keratinocytes; - a dermis of human fibroblasts; and - immune cells derived from human induced pluripotent stem cells.