460results about "Gastrointestinal cells" patented technology

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing intestinal organs and application of the reagent combination or kit. According to the reagent combination or the kit, intestinal organs can be obtained from cell-derived epithelial cells obtained from a donor in a non-invasive manner, and the obtained intestinal organs can be cryopreserved and recovered and can be amplified in vitro for a long time; transcriptome characteristics are similar to those of real human small intestine tissues, and typical marker genes of various small intestine pedigree cell types are highly expressed; compared with intestinal organs obtained through induction of pluripotent stem cells, the intestinal organs have more intestinal pedigree characteristics, and the intestinal function development is more mature; after being promoted to be mature, the intestinal organ also highly expresses genes related to drug absorption and metabolism, has drug absorption capability similar to that of an immortalized intestinal cell line, but more prominently shows intestinal cell lineage characteristics, is closer to an intestinal environment in a real human body, and can be used for screening intestinal disease drugs; the intestinal barrier function is realized.

The invention discloses a 3D intestinal organ differentiation method based on human pluripotent stem cells and an induction culture medium and application thereof, and relates to the technical field of stem cells. According to a culture medium formula combination, intestinal organs can be differentiated into various cell types such as epithelial cells, neuroendocrine cells and endothelial cells; the method is a key mark for successful differentiation and functional maturation of intestinal organs. According to the 3D intestinal organ differentiation method disclosed by the invention, histological structures such as intestinal crypts are differentiated from intestinal organs generated by differentiation, and the intestinal organs can creep in a maintenance stage, so that the intestinal organs are changed from structural bionics to functional simulation, and the significance of the 3D intestinal organ differentiation method is far better than that of pure morphological simulation. Through a systematic culture medium formula, a clear operation process and a multi-stage induction strategy, the 3D intestinal organ with structural integrity, cell diversity and functional activity is successfully constructed, and the system provides an efficient, reliable and extensible in-vitro model platform for intestinal biological research and related application.

The invention relates to the technical field of tumor biomedical engineering and organ chips, in particular to a vascularized gastric cancer organ chip and a preparation method thereof.The chip is composed of an integrated micro-fluidic main body, an annular micro-column array and an optical sealing film, and a central culture cavity is divided into a tumor area and a blood vessel area by micro-columns; the preparation method comprises the following steps: injecting a fibrous protein solution containing human umbilical vein endothelial cells and cancer-related fibroblasts into a vascular region, and adding thrombin for in-situ gelation to form a pre-vascularized network; the method comprises the following steps: mixing a patient-derived gastric cancer organ with a methacrylic acid esterified gelatin pre-polymerized solution, injecting the mixture into a tumor area, and carrying out photo-crosslinking immobilization; culturing for 7-14 days under the dynamic perfusion condition of a mixed culture medium to obtain a three-dimensional gastric cancer organ model containing a capillary network; the method is expected to be used for rapid, low-cost and high-throughput screening of chemotherapy or anti-angiogenesis drugs and accurate prediction of individual curative effects of patients.

The invention discloses an inflammatory bowel disease model, a construction method and application thereof and a drug evaluation method based on the inflammatory bowel disease model, and belongs to the technical field of inflammatory bowel disease model construction. A highly bionic intestinal crypt-villus three-dimensional structure is formed based on self-organization of an intestinal cell line C2BBe1, the intestinal crypt-villus three-dimensional structure and a DMEM complete medium containing dextran sulfate sodium are added into an upper chamber of a double-layer culture chamber Transwell, the DMEM complete medium is added into a lower chamber of the double-layer culture chamber Transwell, culture is performed for 2 days, and the inflammatory bowel disease model is obtained. The method takes effect quickly and is highly simulated, the obtained model can simulate the space-time dynamic development process of intestinal inflammation molecular characteristics, and the natural recovery model and the drug treatment model constructed based on the model can provide a brand new platform for researching an inflammation regression mechanism and a drug intervention effect.

The present invention relates to inflammatory bowel disease model production using patient-derived intestinal organoids, and use thereof, wherein the inflammatory bowel disease model produced by an optimized production method for inflammatory bowel disease model production can be used as a patient-customized bowel disease model by constructing an evaluation method for inflammatory bowel disease drugs.

The invention relates to the technical field of cell biology, in particular to a construction method and application of intestinal organs for children with Crohn's disease, the intestinal organs for children with Crohn's disease are successfully constructed by utilizing intestinal tissues from children with Crohn's disease, and the intestinal organs have various characteristic cells of the intestinal tissues; besides, immune cells are extracted from peripheral blood of a child patient homologous with intestinal tissues, a co-culture system is established with the organ-like model, the intestinal immune microenvironment of the child patient suffering from the Crohn's disease can be reproduced, and the system provides a new model for related mechanism exploration and drug screening of the child Crohn's disease.

The invention discloses a construction method and application of a gastric cancer treatment effect evaluation model of CAR-T cells, and the constructed gastric cancer treatment effect evaluation model of the CAR-T cells comprises a CAR-T cell and gastric cancer organ co-culture killer dynamic detection system and a CAR-T cell, intestine organ and lung organ co-culture safety evaluation system. Based on a CAR-T cell and gastric cancer organoid co-culture lethality dynamic detection system, the process that CAR-T cells act on gastric cancer organoid can be dynamically observed through a living cell imaging method, then the action mechanism of the CAR-T cells for treating solid tumors is studied, and meanwhile, based on a CAR-T cell, intestine organoid and lung organoid co-culture safety evaluation system, the CAR-T cells and the gastric cancer organoid co-culture lethality dynamic detection system can be used for evaluating the CAR-T cells and the gastric cancer organoid co-culture lethality. Compared with the prior art, the preclinical model has the advantages that the effect of the CAR-T cells on gastric cancer para-tissues and other tissue-derived organs can be detected, and safety evaluation is carried out, so that the preclinical model for more comprehensively detecting the gastric cancer treatment effect of the CAR-T cells is obtained, and the research on CAR-T cell targets is facilitated.

The present invention relates to a method for constructing a gene network through a single-cell transcriptome and a method for discovering key genes in differentiation using same, and a composition for the prevention, alleviation, or treatment of colon cancer using the target discovered through the method. The method for constructing a gene network of key genes according to the present invention employs single-cell transcriptome data and thus can be applied to all single-cell transcriptome data. The combination of MYB / HDAC2 / FOXA2 discovered upon application to colon cells can serve as a cancer treatment target that promotes the differentiation of colon cancer cells to revert same into differentiated normal cells.

The invention provides a method for separating, culturing and activating NK (Natural Killer) cells and an application method of the NK cells in tumor immunity. The method comprises the following steps: separating mononuclear cells from peripheral blood of healthy adults, removing non-NK cell populations through a negative selection method, and enriching and purifying NK cells. Through stimulation of specific cytokines, NK cells are successfully amplified, and cytotoxicity of the NK cells is activated. By optimizing the preparation and activation method of the NK cells, the NK cells with efficient killing activity are obtained in vitro, and the tumor treatment effect is remarkably improved through combined use of the NK cells and the immune checkpoint inhibitor. In combination with the immune memory effect of the NK cells, the invention provides a new anti-tumor immunotherapy method which can be applied to preparation of new anti-tumor drugs and provides an innovative thought for tumor treatment.

The present invention discloses a buffer for tissue separation and / or preservation and its application, and a method for culturing tumor organoids, and relates to the field of organoid culture. The buffer is used to preserve tumor tissue samples, which are of liquid origin; the buffer includes: a basal culture medium and specific additive factors, and the specific additive factors include: superoxide dismutase, catalase, penicillin, and streptomycin; based on the volume of the basal culture medium, the concentration of superoxide dismutase is 1 to 100 μg / mL, the concentration of catalase is 50 to 500 μg / mL, the concentration of penicillin is 50 to 200 U / mL, and the concentration of streptomycin is 50 to 200 μg / mL. The tissue preservation solution of the present invention is used for the transportation and protection of liquid-derived tumor tissue samples, and has a high tumor cell activity retention rate. It is used to construct tumor organoids with a high success rate and a large number, which can meet the needs of clinical detection and disease research.

The invention belongs to the field of organoid culture, and particularly relates to a mixed type gastric cancer organoid culture medium and a culture method and application thereof. Wherein the culture medium comprises a basic culture medium and a specific additive factor; the culture method comprises the following specific steps: treating mixed gastric cancer tissues with tissue digestive juice to obtain mixed gastric cancer primary cells, and culturing with a mixed gastric cancer organ culture medium to obtain mixed gastric cancer organs. The mixed type organ-like form corresponding to the mixed type gastric cancer tissue can be cultured by using the mixed type gastric cancer organ-like culture medium disclosed by the invention. By means of methods such as organoid passage and cryopreservation technologies, the culture success rate of the mixed type gastric cancer organoid is high, sufficient mixed type organoids can be obtained within limited time, an accurate tumor model is provided for individualized tumor therapy, and valuable samples are preserved for scientific research.

The invention discloses a culture system, culture method and application of gastrointestinal tumor chemotherapy drug-resistant organoid, the culture system comprises: a basic culture medium, which is a DMEM / F12 culture medium, and is additionally added with a special growth promoting factor obtained from a natural extract; the multiple growth factors comprise an epidermal growth factor (EGF), a fibroblast growth factor (FGF), a bone morphogenetic protein (BMP) and a vascular endothelial growth factor receptor inhibitor (VEGFRi), and the concentration of each growth factor is accurately screened and optimized; the extracellular matrix is Matrigel modified by physical and chemical methods; the specific additive comprises an antioxidant N-acetylcysteine (NAC). According to the culture system, the culture method and the application of the gastrointestinal tumor chemotherapy drug-resistant organoid, the culture system which is closer to a real environment in vivo is constructed by optimizing basic culture medium components, setting various growth factors and specific additive concentrations and adopting modified Matrigel as an extracellular matrix.

The present invention relates to a method for preparing organ-specific mesenchymal cells from pluripotent stem cell-derived intestinal organoid stromal cells. By using cells derived from stromal cell layers adjacent to intestinal organoids for differentiation into organ-specific mesenchymal cells, the present invention can greatly increase the efficiency of differentiation into stromal cells through the regulation of retinoic acid (RA) and hedgehog (HH) signaling pathways, and can increase the expression of organ-specific markers without exhibiting undifferentiated state cell characteristics, and thus mesenchymal cells having well-simulated biological characteristics can be prepared.

The invention discloses a 3D (three-dimensional) visual bionic intestine-blood vessel-nerve interface organ simulation chip, which is characterized in that a micro-fluidic chip is used as a platform to construct an intestinal tract-blood vessel-brain system which simulates biological transformation of perfluorinated compounds in intestinal tracts under near physiological conditions and biologically transports the perfluorinated compounds through blood vessels; a neuronal cell culture chamber, a vascular endothelial cell culture chamber and an intestinal tract cell culture chamber are respectively arranged on the micro-fluidic chip, the culture chambers are communicated through micro-columns, and the vascular endothelial cell culture chamber serves as a barrier in the middle; perfluorinated compounds enter corresponding cell culture cavities for culture through reagent injection channels of different cell culture channels, generated metabolites are discharged through the channels, and multi-organ interaction and toxicity induced by the perfluorinated compounds are analyzed. The chip can simulate an intestinal barrier structure, a blood vessel and a neural network structure, can highly simulate biological transport and transformation of a perfluorinated compound between intestines and brains, and effectively reduces cost and time.

The invention belongs to the technical field of biological medicine, and particularly relates to a construction method of a gastric mucosa intestinal epithelial metaplasm type organ model with goblet cell phenotype. The model is characterized by MUC2 + MUC6 + goblet cells, and histological characteristics of gastric mucosa intestinal metaplasia are more deeply simulated from the aspect of cell phenotype. The construction method comprises the following steps: (1) obtaining a human stomach gland to prepare an organoid; (2) subculturing the organoid; (3) construction of a pCDH-CMV-MCS-EF1-CDX2-Puro overexpression lentiviral vector, and construction of a pCDH- (4) virus packaging; (5) constructing CDX2 overexpression stably transfected cells and organoid; and (6) carrying out gradient differentiation on the CDX2 overexpression organoid to prepare the intestinal metaplasia organoid. According to the invention, the pathological transformation of the cytology level in the gastrointestinal mucosa intestinal epithelial metaplasia process is realized in the in-vitro model for the first time, and a new research model and theoretical guidance are provided for the diagnosis, prevention and individualized precise treatment of precancerous lesions of gastric cancer.

The invention relates to an establishment method and application of in-vitro intestinal inflammation models of mice of different ages, and relates to the technical field of biology. According to the invention, young, adult, middle-aged and old wild type (WT) and TLR4 gene deletion type (TLR4 <- / ->) mice are selected, colon tissues are cultured in vitro, inflammatory response is induced by using interleukin-1beta (IL-1beta) stimulation, an age-related intestinal inflammation model is constructed, the inflammation phenotype of the mouse model and the activity of a TLR4 / MyD88 / NF-kappa B signaling pathway are determined by comparing WT and TLR4 <- / ->, and the age-related intestinal inflammation model is constructed. The core function of the TLR4 in regulating age-dependent intestinal barrier dysfunction is defined, a regulatory network of the TLR4 in intestinal immune aging and flora-host interaction is disclosed, and an age personalized medical strategy is promoted. According to the invention, an efficient and customizable multi-age in-vitro intestinal inflammation model is constructed, and an innovative platform is provided for mechanism analysis and intervention drug screening of TLR4 targeted therapy and age-related intestinal diseases.

The invention discloses a rectal cancer organoid culture kit and a culture method thereof, and particularly relates to the technical field of genetic engineering.The kit comprises a culture medium with specific components, a sample treatment reagent, a passage reagent and a preservation reagent, and the culture medium contains Advanced DMEM / F-12 and multiple factors; the sample treatment reagent can effectively treat tissues; a passage reagent guarantees passage and cryopreservation; and the preservation reagent ensures the cell cryopreservation and resuscitation effects. The culture method covers scaffold and scaffold-free culture processes and comprises the steps of primary culture, subculture, recovery and the like, and the steps are operated finely. The method has the advantages of obvious advantages, fast tissue dissociation, small damage, good cell suspension dispersion, no DMSO toxicity, simple culture, high amplification efficiency, stable passage, wide applicability and high cost performance, and can provide a powerful tool for rectal cancer research, drug screening and the like.

The invention belongs to the technical field of biology, and particularly relates to a construction method and application of a model with fusobacterium nucleatum damaged mouse intestinal tract organoid, and the construction method comprises the following steps: separating mouse intestinal tract organoid; obtaining a culture supernatant with fusobacterium nucleatum; obtaining a cell-free extract with fusobacterium nucleatum; viable bacteria with fusobacterium nucleatum are obtained; and treating the mouse intestinal organ with the fusobacterium nucleatum culture supernatant or the fusobacterium nucleatum cell-free extract or the fusobacterium nucleatum viable bacteria to obtain the model of the mouse intestinal organ damaged by the fusobacterium nucleatum. The construction method of the organ-like model with the fusobacterium nucleatum damaged intestinal tract is simple, operability and convenience can be increased for further research on a regulation mechanism of the fusobacterium nucleatum damaged intestinal tract, and a reliable and accurate research tool is provided for gastrointestinal diseases such as inflammatory bowel diseases and colorectal cancer.

The invention discloses a novel colorectal cancer organoid culture method, which obviously improves the activity of primary cells (more than or equal to 90%) and the forming efficiency of the organoid (the success rate is more than 95%) through alcohol gradient cleaning, collagenase-hyaluronidase combined digestion and stent-free culture technologies. The improved culture medium takes R-Spondin1 as a core, nicotinamide and a small-molecule inhibitor are combined, and long-term amplification of organoids (passage is more than or equal to 10 times) is supported. The method is suitable for individualized drug screening, tumor microenvironment simulation and drug resistance mechanism research, has the advantages of high efficiency, stability and low cost, and provides a reliable model for precise medical treatment of colorectal cancer.

The invention discloses a perfusion-free parallel double-cell intestine-liver chip system and application thereof in drug uptake evaluation, and belongs to the field of biomedical engineering and organ chips. The chip disclosed by the invention is simple in structure, free of an external pump and a complex pipeline, capable of realizing fluid circulation through swinging, convenient to operate, low in cost and small in pollution risk, and also capable of promoting cell differentiation and optimizing hepatocyte functions. The chip integrates a parallel intestinal cell and a liver cell which are communicated through a shared channel, can continuously simulate intestinal absorption and liver first-pass metabolism, and is closer to an in-vivo physiological state. The device adopts a detachable transparent modular design, can be repeatedly used, supports multi-group parallel operation, can improve experimental flux and result repeatability, and is suitable for in-vitro verification of drug absorption, permeability and metabolism evaluation.

The invention provides a preparation method of a fragmented tumor antigen. The preparation method comprises the following steps: obtaining a tumor cell suspension; and irradiating and incubating the tumor cell suspension to prepare the fragmented tumor antigen. The preparation method is simple, low in cost and convenient to store, the fragmented tumor antigen prepared through the preparation method can remarkably inhibit tumor growth in vivo after being inoculated to an individual, tumor metastasis and relapse can also be effectively inhibited, immune memory is induced to be generated, the continuous anti-tumor effect is achieved, and the preparation method is suitable for clinical application. The control on metastatic tumors is obviously improved. The effective anti-tumor effect can be achieved in vivo when the fragmented tumor antigen prepared by the invention is independently or jointly used.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the second leading cause of cancer death worldwide. Novel tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs) expressed by colorectal tumor cells are described herein. Synthetic long peptides, nucleic acids, compositions, cells, TCRs, antibodies and vaccines derived from these TSAs and TAAs are described. The use of the TSAs / TAAs, nucleic acids, compositions, antibodies, cells and vaccines for the prevention or treatment of cancer expressing the TSAs and / or TAAs, including CRC, is also described.

Provided is a cell culture substrate comprising a substrate and a layer that covers at least a portion of the surface of the substrate, has a layer thickness of 5 to 2000 nm and contains a hydrophilic polymer, in which the hydrophilic polymer contains a phosphorylcholine group or a hydroxyl group, the cell culture substrate has a region (A) and a region (B) as mentioned below, and the bump height in a boundary region between the region (A) and the region (B) is 1 to 500 nm. (A) An island-like region which has cell adhesiveness and cell proliferating properties and has an area of 0.001 to 5 mm2. (B) A region which is located adjacent to the region (A) and does not have cell adhesiveness or cell proliferating properties.

The invention belongs to the field of biology, and discloses a cdc42 gene knockout HCT-8 cell line and application thereof, the taxonomic name of the cdc42 gene knockout HCT-8 cell line is Homo sapiens, the cdc42 gene knockout HCT-8 cell line is preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC NO: 66616; the preservation date is July 01, 2025; the preservation address is the fifth floor of building 59, No.100 courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province. According to the invention, a cdc42 gene in an HCT-8 cell is knocked out by adopting a CRISPR-Cas9 technology so as to construct a cdc42 gene knocked-out HCT-8 cell line, and the cdc42 gene knocked-out HCT-8 cell line is used as a cryptosporidium parvum cell infection model.

The invention provides a method for evaluating drug enterotoxicity in vitro through an intestinal organ model, and relates to the field of biological medicine. Comprising the following steps: preparing six different culture media, culturing the hESC by using the different culture media, inducing the hESC to respectively perform directional endoderm cell differentiation, posterior intestinal bulb differentiation, intestinal organ differentiation and intestinal organ maturation, and evaluating the drug enterotoxicity by using a prepared intestinal organ model. The intestinal organ model prepared by the invention is similar to human intestinal cells in composition, can accurately reflect the toxic effect of the medicine in the physiological environment in the human body, can be repeatedly obtained in quantity, and avoids clinical risks and ethical constraints; the sensitivity of drugs with different enterotoxicity can be reflected through cell viability, and the method is suitable for rapid evaluation of in-vitro high-flux drug enterotoxicity.

Disclosed are compositions, in particular, organoid compositions, derived from more than one donor cell. Further disclosed are methods of making compositions, for example, organoid compositions, that comprise a differentiated cell population derived from more than one donor cell. Donor cells may include, for example, a precursor cell such as an embryonic stem cell or other precursor cell. The disclosed methods use synchronization conditions to produce a synchronized pooled-precursor cell population, which may then be differentiated into an organoid composition. Methods of using the compositions are also disclosed.

The application discloses application of a pharmaceutical composition in treating cancer, wherein the pharmaceutical composition comprises a BCL-XL inhibitor and a HER2 targeted drug or comprises a BCL-XL inhibitor and a STAT3 inhibitor. The application provides medical use of the pharmaceutical composition according to the application for treating or preventing cancer. The application also provides a method for treating or preventing cancer, and provides a brand-new treatment scheme for the technical field of cancer treatment, and has important scientific significance and clinical application value.

The invention discloses a medicine screening method based on colorectal cancer organoid, which comprises the following steps: adding a colorectal cancer culture medium into the cultured colorectal cancer organoid, culturing for 48-72 hours, adding a to-be-detected colorectal medicine, continuously culturing for 72-96 hours, detecting the cell activity, and screening the medicine capable of reducing the cell activity. According to the method, the tumor microenvironment of the organoid is utilized, the modeling period is shortened, the cost is reduced, the tumor state can be simulated, high-throughput drug screening can be carried out in vitro, and the method has important significance on accurate treatment of lung cancer.

Methods for computing scores, training models, and analyzing data in gene expression and infection studies are disclosed. Methods include receiving and processing data, generating scores, and displaying results. Systems and mediums for executing these methods are also disclosed, enabling insights into gene expression dynamics and infection progression.

The invention provides an application of recombinant III-type humanized collagen in preparation of a medicine for preventing and / or treating gastric cancer, which can overcome the defect that the existing medicine generates more adverse reactions in treatment.