296results 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 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 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.

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.

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.

The invention discloses an application of knocking down intestinal epithelial cells Mysm1 in increasing secretion of 5-HT by the intestinal epithelial cells. The invention provides application of a substance capable of inhibiting Mysm1 gene expression and / or activity in promoting intestinal tract / intestinal epithelial cells to secrete 5-hydroxytryptamine. According to the invention, the synthesis amount of the 5-HT in the intestinal tract can be manually regulated and controlled, so that the functions of the 5-HT in intestinal brain communication, intestinal tract barrier and the like are regulated and controlled; a product for promoting intestinal epithelial cells to secrete 5-hydroxytryptamine can be prepared, so that the 5-HT function is productized, and the use is convenient.

The invention relates to the technical field of biology, in particular to a composition, a culture medium and application of the culture medium in intestinal organ culture. The glucosamine hydrochloride is applied to culture of intestinal organs for the first time, organ proliferation and differentiation can be effectively promoted, and the number of formed organs is remarkably increased. On the basis, a three-dimensional culture system for staged time sequence induction in a postpartum newborn period, a child period and an adult period is constructed, and accurate construction of intestinal organs of the mice in different development periods is realized by combining dynamic regulation and control of an extracellular matrix and combinatorial optimization of growth factors; and an in-vitro model covering multiple time sequence stages is provided for intestinal development mechanism research and disease modeling.

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.

The invention relates to a cellulose-based hydrogel prepared based on a cellulose copolymer, a cellulose derivative copolymer or a blend thereof, ultramicro fibers of cellulose form a net structure, and the cellulose-based hydrogel has a highly-ordered three-dimensional micro-nano structure, is better in transparency and is more beneficial to observation of a cell state. The cellulose-based hydrogel is suitable for culture of 3D cell culture tissues and organoids, the stem cells cultured by the cellulose-based hydrogel can form cluster culture stem cells and organoids, the operation is simple, compact fused monolayer endoderm cells are formed, non-directional differentiation of the stem cells can be inhibited, and the formed organoids are thick in vesicle wall and are in a solid shape. And the obtained product is a differentiated mature organ and can be used for high-throughput drug screening, drug toxicity testing and regenerative medicine.

The generation of complex organ tissues from human embryonic and pluripotent stem cells (PSCs) remains a major challenge for translational studies. It is shown that PSCs can be directed to differentiate into intestinal tissue in vitro by modulating the combinatorial activities of several signaling pathways in a step-wise fashion, effectively recapitulating in vivo fetal intestinal development. The resulting intestinal “organoids” were three-dimensional structures consisting of a polarized, columnar epithelium surrounded by mesenchyme that included a smooth muscle-like layer. The epithelium was patterned into crypt-like SOX9-positive proliferative zones and villus-like structures with all of the major functional cell types of the intestine. The culture system is used to demonstrate that expression of NEUROG3, a pro-endocrine transcription factor mutated in enteric anendocrinosis is sufficient to promote differentiation towards the enteroendocrine cell lineage. In conclusion, PSC-derived human intestinal tissue should allow for unprecedented studies of human intestinal development, homeostasis and disease.

The invention provides for methods of differentiating pancreatic endocrine cells into pancreatic beta cells expressing PDX1, NKX6.1, MAFA, UCN3 and SLC2A. These pancreatic beta cells may be obtained by step-wise differentiation of pluripotent stem cells. The pancreatic beta cells exhibit glucose-dependent mitochondrial respiration and glucose-stimulated insulin secretion similar to islet cells.

The invention discloses application of ceramide and a synthetic regulating agent thereof in regulating porcine epidemic diarrhea virus infection, and belongs to the technical field of biology. Experiments prove that the mRNA expression quantity and virus titer of PEDV (porcine epidemic diarrhea virus) in IPEC-J2 cells are remarkably improved by exogenously adding C24-ceramide or regulating and controlling the endogenous ceramide level through a ceramide enzyme inhibitor, and a ceramide synthesis inhibitor shows a remarkable antiviral effect. The invention proves that ceramide is a key factor for PEDV (porcine epidemic diarrhea virus) replication, and an inhibitor targeting a ceramide synthetic route can effectively inhibit PEDV infection, so that a theoretical basis and an experimental basis are provided for constructing a porcine epidemic diarrhea virus susceptibility model and developing a novel anti-PEDV drug.

The purpose of the present invention is to provide a novel method for culturing tissue-derived cells. The method uses a cell culture method including a step for co-culturing cells derived from the skin and at least one type of cell selected from among cells derived from the intestine, cells derived from the heart, cells derived from the brain, cells derived from the stomach, cells derived from the embryonic membrane, cells derived from the kidney, and cells derived from the lung. And culturing the skin-derived cells.

The invention provides for methods of differentiating pancreatic endocrine cells into pancreatic beta cells expressing PDX1, NKX6.1, MAFA, UCN3 and SLC2A. These pancreatic beta cells may be obtained by step-wise differentiation of pluripotent stem cells. The pancreatic beta cells exhibit glucose-dependent mitochondrial respiration and glucose-stimulated insulin secretion similar to islet cells.

A cell culture substrate according to the present disclosure is a cell culture substrate including at least one well including a side surface and a bottom surface, in which the side surface includes a first side portion defining a first space and a second side portion defining a second space communicating with the first space, and in plan view, an upper end portion of the first side portion is formed in a C shape having a central angle of 240 degrees or more, the second space extends from an opening portion of the upper end portion in a first direction away from the first space in the plan view, and the second side portion includes a facing surface facing the opening portion in the first direction, the facing surface being disposed at a position where a maximum distance between the opening portion and the facing surface in the first direction is smaller than a diameter of the C shape of the upper end portion in plan view.

Disclosed herein are enteroids, obtained by dissociating an epithelial cell population from a non-transplanted human intestinal organoid (HIO), such as an HIO produced in vitro from human pluripotent stem cells (hPSCs). Also disclosed herein are methods of producing and using the same, including methods of transplantation involving said enteroids, particularly for treating a gastrointestinal disease or condition. Also disclosed herein are methods of transplantation involving HIOs and / or HIO components, particularly for treating a gastrointestinal disease or condition.

The invention relates to the technical field of biology, and discloses a construction method of a gastric cancer organoid culture system, which comprises the following steps: preparation of a conditioned medium: culturing L-WRN cells and collecting the conditioned medium; preparing a gastric cancer organoid culture medium, and mixing the conditioned culture medium, the basic culture medium, a plurality of growth factors and additives; and extraction and culture of organoid: separating cells from gastric cancer tissues, mixing the cells with matrigel, and culturing by using the prepared culture medium. According to the method, the repeatability and the stability of the organoid model are improved, the response consistency of the organoid in drug screening is improved, and the reliability of the model in disease mechanism research and personalized medical application is guaranteed.

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 invention belongs to the technical field of biology, and discloses a preparation method of an intestinal plexus patch, which comprises the following steps: killing an intestinal tract donor, fixing intestinal tissue segments of duodenum, jejunum, ileum or colon by using paraformaldehyde, and dehydrating to obtain dehydrated intestinal tissue segments; washing the dehydrated intestinal tissue section and removing mesentery, fat and Pari's lymph nodes attached to the jejunum, cutting an intestinal mass of about 4-6mm perpendicular to the intestinal tissue section, flatly laying the intestinal mass on a glass slide, dropwise adding a PBS (Phosphate Buffer Solution) of Triton X-100, fixing the intestinal mass by using a smooth surface of a pair of tweezers, stripping muscle tissues along the annular muscle direction, and drying the intestinal mass in a drying oven. The intestinal slices with intermuscular nerve plexus are obtained; finally, the intestinal piece with the intermuscular nerve plexus is cleaned, then fluorescent staining and piece sealing are conducted, and the mouse intestinal nerve plexus patch is obtained. The preparation method of the mouse intestinal nerve plexus patch has the advantages of being convenient, rapid and the like.

The invention discloses application of a DDIT3 gene as a target spot in regulation and control of bovine coronavirus replication. Experiments find that regulation and control of the expression level of the DDIT3 gene can influence the replication level of the BCoV, when the expression of the DDIT3 gene in a host cell is inhibited, the replication of the BCoV can be promoted and the titer of the virus can be improved, and when the DDIT3 is over-expressed on the host cell, the replication of the BCoV can be inhibited and the titer of the virus can be reduced. Experimental results show that the DDIT3 gene can be used as a novel target for developing BCoV antiviral drugs, and a tool and a material are provided for researching a molecular mechanism of the DDIT3 gene for regulating and controlling pathogenic microorganism replication in cells.

The invention discloses a human-derived ampulla cancer cell line DPC-X5 and application thereof, and belongs to the field of microbial animal cell lines. The human-derived ampulla cancer cell line is named as a human-derived ampulla cancer cell line (homo sapiens) DPC-X5, and is preserved in the China Center for Type Culture Collection on October 30, 2025, and the preservation number is CCTCC NO: C202580. The human-derived ampulla cancer cell line DPC-X5 can be applied to establishment of a cell model for occurrence, development or metastasis of ampulla cancer. The human-derived ampulla cancer cell line DPC-X5 can be applied to cell models for researching differentiation mechanisms, cell morphology and dysfunction and tumor infiltration and metastasis mechanisms of ampulla cancer and guiding clinical comprehensive diagnosis and treatment. The human-derived ampulla cancer cell line DPC-X5 can be applied to research on the occurrence mechanism of the ampulla cancer and screening of drugs for preventing and treating the ampulla cancer. The human-derived ampulla cancer cell line DPC-X5 can be applied to establishment of an ampulla cancer animal model.

Disclosed herein are improved in vitro methods of making splanchnic mesoderm cell types and subtypes thereof from pluripotent cells. These methods can be used to produced improved foregut- and hindgut-derived organoids containing enriched mesenchyme, which enhances organoid viability, growth, and maturation, both in in vitro culture and in vivo transplantation.