217results about "Coculture" patented technology

The invention provides a method for drug screening and disease model construction of liver organoid, relates to the technical field of medical research, and adopts multi-modal data acquisition, fusion and intelligent analysis and quantum computing processing to optimize comprehensive application of data, so that the richness, accuracy and analysis efficiency of the data are remarkably improved, and the risk of drug screening and disease model construction of liver organoid is reduced. Multi-source data of genomics, transcriptomics and the like are collected comprehensively, an accurate research system of immune-hepatic disease interaction based on organoid is constructed, an in-vivo complex immune-hepatic disease microenvironment can be highly simulated, the proportion and activation state of immune cells are accurately regulated and controlled in an in-vitro model, functional and phenotypic changes of the immune cells are monitored in real time, and the immune-hepatic disease interaction accuracy is improved. According to the present invention, the interaction mechanism between the immune cells and the liver organ cells is deeply revealed, such that the accurate information basis is provided for the accurate determination of the drug action target, the development of the innovative drug for the immune check point or tumor microenvironment regulation is easily achieved, and the pertinence and the success rate of the drug research and development are substantially improved.

The invention discloses a vascularized immune co-culture organoid chip for simulating a tumor microenvironment. The vascularized immune co-culture organoid chip comprises a chip main body and a bottom plate, a flow channel and a liquid storage tank are arranged in the chip main body; the runners comprise a first runner, a second runner, a third runner and a fourth runner; a compartment is arranged in the middle of the third flow channel; the first flow channel is connected with a first liquid storage tank which is used for injecting a first hydrogel suspension obtained by mixing fibroblasts and hydrogel into the first flow channel; the second flow channel is connected with a second liquid storage tank for injecting a culture solution into the second flow channel; a compartment is arranged in the middle of the third flow channel; the third flow channel is connected with a third liquid storage tank communicated with the compartment, and the compartment is connected with a compartment liquid storage tank; the compartment liquid storage tank is used for injecting a second hydrogel suspension obtained by mixing tumor organs and hydrogel into the compartment, and the third liquid storage tank is used for injecting hydrogel into the third flow channel; and the fourth flow channel is connected with a fourth liquid storage tank for injecting vascular endothelial cell and immune cell suspension into the fourth flow channel. The invention further discloses a vascularized immune co-culture organoid culture method for simulating the tumor microenvironment.

The invention provides a microfluidic chip, system and method for osteochondral organs, and relates to the technical field of organoid microfluidic chips, the chip comprises a bone microvascular endothelial cell channel, a bone compartment, a cartilage compartment and a synovial fluid channel which are arranged in parallel and in sequence, and permeation areas between the channel and the compartments and between the compartments. According to the scheme, the complex structure and function of a human osteochondral system are effectively simulated, the physiological and pathological processes of human bone joints can be simulated more truly in an in-vitro environment, and a method for establishing a highly-simulated femoral head necrosis model and osteoarthritis model based on the chip is provided. The problems that a traditional model is insufficient in physiological environment simulation, lack of control over dynamic culture conditions, tedious and time-consuming in the construction process, difficult to support high-throughput experiments and the like are solved.

The invention relates to the technical field of biological tissue engineering, and discloses a nerve-blood brain barrier co-culture model, which comprises a culture chip, and nerves and blood brain barriers respectively constructed in upper culture micropores and lower culture micropores of the culture chip. Wherein a semi-permeable membrane is arranged between the lower-layer culture micropores and the upper-layer culture micropores of the culture chip; the nerves are obtained by inoculating neurons / brain organs obtained by stem cell differentiation into the upper culture micropores for co-culture, and the blood brain barrier is obtained by inoculating brain microvascular endothelial cells obtained by stem cell differentiation into one side of the semipermeable membrane in the lower culture micropores for co-culture. The co-culture model is higher in biomimetic property, and can better simulate the structure and function of the blood brain barrier, for example, the co-culture model has a barrier function closer to the physiological level, the selective permeability of a small molecule drug is matched with the in-vivo permeation condition, and the co-culture model has the function of a small molecule related transporter. The invention further discloses a construction method and application.

The invention relates to a culture medium and a culture method of persistent DC-CIK cells and application of the culture medium and the culture method, and belongs to the technical field of cell culture. The invention provides a culture medium for persistent DC-CIK cells, which takes a serum-free culture medium as a solvent and comprises a recombinant human granulocyte macrophage colony stimulating factor, a recombinant human interleukin-12, a recombinant human tumor necrosis factor-alpha, a recombinant human interferon-gamma, L-cysteine, glutathione, ginseng polysaccharide and streptomycin sulfate. According to the culture medium and the culture method of the persistent DC-CIK cells, proliferation and growth of the DC-CIK cells can be promoted, and very high tumor killing activity of the DC-CIK cells can be maintained for a long time.

The invention relates to a cell collection method capable of simultaneously collecting three cells in a co-culture model, and belongs to the technical field of biology. The invention provides a cell collection method capable of simultaneously collecting three cells in a co-culture model, and the cell collection method comprises the following steps: after a three-cell co-culture model is constructed, taking out a Transwell chamber, retaining cells in the lower chamber, and collecting the cells in the lower chamber; respectively digesting the cells on the two sides of the Transwell cell membrane by using a trypsin solution with the concentration of 0.5 g / 100mL so as to respectively collect the cells on the two sides of the Transwell cell membrane. According to the cell collection method disclosed by the invention, the three cells in the three-cell co-culture model are simultaneously collected in a manner of digesting the cells on the two sides of the Transwell membrane step by step by using pancreatin, so that not only is the cell and consumable cost saved, but also the experimental synchronism of the three cells is ensured, and convenience is provided for optimizing the experimental process.

The present invention belongs to the field of biomedical engineering technology and relates to a bone organoid, its construction method, and its use. The bone organoid is obtained by three-dimensionally culturing a cell mixture comprising stem cells and macrophages. The bone organoid and its construction method of the present invention enable rapid preparation of bone organoids with minimal differences from natural bone, enabling regulation of cell fate and better bone formation and repair.

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 relates to a pathological pulmonary fibrosis model and a construction method and application thereof. The pathological pulmonary fibrosis model provided by the invention is good in stability and controllability and high in maturity, not only can better simulate the function and structural characteristics of pulmonary alveoli-capillary barrier, but also has the pathological characteristics and immune microenvironment of pulmonary fibrosis lesions, has the advantage of high simulation degree, and has good application prospects in simulation and research of pulmonary fibrosis diseases. Good application prospects are realized in the aspects of drug detection, screening, evaluation, research and development and the like.

The invention discloses a bone organ chip loaded with a microelectrode array and a use method, and relates to the technical field of organ chips, the bone organ chip is made of a polydimethylsiloxane material, and the bone organ chip comprises a three-layer structure: a top layer, a middle layer and a bottom layer; the middle layer comprises two blood vessel micro-channels, two osteoblast micro-channels and a neuron micro-channel, the channels are separated through columns and micro-columns, and cell interaction in a bone micro-environment is simulated; a microelectrode array is arranged on the top layer and is used for monitoring electrical activity of neurons in real time; the bottom layer is provided with a pneumatic valve which can control opening and closing of the micro-channel. According to the invention, nerve and blood vessel channels are introduced, a nerve-blood vessel-immunoregulation bone microenvironment is constructed, physiological processes such as osteogenesis-blood vessel formation coupling and the like are embodied, intervals of different sizes can enhance intercellular crosstalk and allow blood vessels and neuron axons to selectively pass through, so that the chip can simulate a complex microenvironment of bone tissues, and the microenvironment of the bone tissues can be simulated; and an efficient and accurate in-vitro model is provided for bone disease mechanism research and drug development.

The invention provides a culture system for simulating neutrophile granulocyte extracellular trap net intervention oligodendrocyte in vitro, and belongs to the technical field of cell and function detection. The invention establishes a stable and repeatable research system by screening and determining the optimal condition for in-vitro co-culture of the neutrophil extracellular trap net and the oligodendroglia. Based on the condition, the experiment can observe the obvious change of the oligodendroglia after the intervention, and the result shows that the condition can accurately simulate the endoplasmic reticulum stress regulation effect of the neutrophile granulocyte extracellular trap net on the oligodendroglia, thereby providing a necessary experimental basis for the subsequent mechanism research.

The invention relates to a retina nerve and blood vessel unit in-vitro three-culture model and a preparation method thereof, and belongs to the technical field of biology. The invention provides an in-vitro three-culture model of a retina nerve and blood vessel unit. The in-vitro three-culture model is obtained by carrying out in-vitro Transwell indirect co-culture on retina microvascular endothelial cells, retina ganglion cells and retina Muller cells. The in-vitro three-culture model of the retina nerve and blood vessel unit is based on a three-cell indirect co-culture method, retina microvascular endothelial cells, retina ganglion cells and retina Muller cells are simultaneously incorporated by utilizing Transwell, and the three cells are in indirect contact. The in-vitro three-culture model of the retina nerve and blood vessel unit can be used for more objectively simulating pathophysiological states of various cells of the retina nerve and blood vessel unit, so that common pathophysiological changes of mutual dependence and interaction of the three cells under in-vitro conditions can be observed.

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 invention provides a stem cell exosome for overexpressing A beta degrading enzyme as well as a preparation method and application of the stem cell exosome. Specifically, the invention provides an application of the stem cell exosome overexpressing A beta degrading enzyme in preparation of drugs, and the drugs are used for at least one of the following: prevention, improvement or treatment of cognitive impairment related diseases; neuronal cell damage is improved; wherein the stem cells for overexpressing the A beta degrading enzyme are prepared through lentivirus transfection. According to the stem cell exosome overexpressing the A beta degrading enzyme, the content of the A beta degrading enzyme is obviously higher than that of the exosome prepared by a traditional method. Therefore, when the compound is prepared into a medicine for treating or preventing cognitive impairment related diseases, the medicine can be efficiently conveyed to the brain, and the concentration of the medicine in the brain is remarkably improved, so that the curative effect is enhanced, the dosage of the medicine can be reduced, the side effect is reduced, and the prognosis of brain diseases is effectively improved.

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

The present invention relates to dedifferentiated plant cells of a plant of the genus Paeonia, or an extract thereof, obtained from at least one plant part selected from the group consisting of buds, petals, leaves and mixtures thereof, and also to a method for obtaining said cells, to cosmetic compositions comprising them, to their use, and to a method for preparing said cells. And cosmetic treatment methods comprising applying the composition to the skin to improve and / or enhance barrier function, to improve moisturizing of the skin, and to act as an antioxidant.

Several embodiments disclosed herein relate to methods and compositions for enhanced expansion of NK cells in culture, through the repeated co-culturing of NK cells with feeder cells and the selective use of stimulatory interleukins. In several embodiments, the methods utilize one or more soluble interleukins as culture media supplements at one or more time points during expansion of the NK cell, or other immune cell, which results in a highly expanded and highly cytotoxic population of cells, for use in, for example allogeneic cellular immunotherapy.

The invention provides a preparation method of DC cells. The preparation method is especially suitable for rapidly obtaining high-purity and high-functional-activity DC cells. The method comprises the steps of separation from peripheral blood mononuclear cells (PBMC), induction culture, maturation treatment and the like, maturation of the dendritic cells is promoted by introducing an anti-CD47 single-domain antibody or LPS, and the immunocompetence of the dendritic cells is remarkably improved. The method is further applied to preparation of an anti-tumor cell preparation, the capability of activating tumor specific T cells by tumor antigen-loaded DC cells can be remarkably enhanced, and the killing activity of the T cells to tumor cells is improved. The preparation method provided by the invention overcomes the defects in the prior art, has the advantages of high efficiency, safety and stability, and can provide a more reliable cell therapy scheme for tumor immunotherapy.

The invention belongs to the technical field of biomedical engineering and tissue engineering, and particularly relates to an acellular matrix modified bionic micro-fluidic organ chip constructed by 3D printing composite near-field direct writing as well as a preparation method and application of the bionic micro-fluidic organ chip. According to the bionic micro-fluidic organ chip prepared by the invention, the microstructure and physiological similarity of a model are remarkably improved, the problems of insufficient bionic property, single cell and the like at present are solved, and the preparation method is simple, is suitable for batch production and solves the problem of high production cost. The invention provides a new technical direction for the construction of the bionic organ model.

The invention discloses a preparation method of a cochlear organ chip integrated with a blood labyrinth barrier, and belongs to the field of biomedical engineering. The preparation method of the simulated blood labyrinth barrier comprises the following steps: constructing an endothelial cell culture cavity and a pericyte culture cavity, and separating the endothelial cell culture cavity and the pericyte culture cavity through a simulated basement membrane; and respectively inoculating percutaneous cells and endothelial cells into a percutaneous cell culture cavity and an endothelial cell culture cavity, and culturing to obtain the simulated blood lost barrier. The chip can effectively simulate the structure and the function of the BLB, and the barrier integrity of the BLB is evaluated through TEER (trans-epithelial electrical resistance) measurement and an apparent permeability coefficient. The invention also relates to application of the chip in NIHL drug screening, in particular to drug evaluation for oxidative stress induced inner ear organ injury. Through the platform, the protection effect of the candidate drugs on the TBHP-induced oxidative stress injury of the inner ear organs can be evaluated, and a new tool and thought are provided for developing novel NIHL treatment drugs.

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 invention relates to a lung in-vitro model and a construction method and application thereof. The lung in-vitro model provided by the invention is good in stability and controllability and high in maturity, can better simulate functions and structural characteristics of pulmonary alveoli-capillary barriers, has the advantage of high simulation, and has a good application prospect in the aspects of disease simulation, drug detection, screening, evaluation, research and development and the like.

The invention provides a construction method and application of a gene modified K562 cell line, and belongs to the technical field of biological medicines. According to the invention, two viruses CD86-4-1BBL and mbIL-21 are respectively subjected to gene transduction packaging by adopting a third-generation lentivirus four-plasmid packaging system, a K562 cell line is infected by the lentiviruses CD86-4-1BBL and mbIL-21, and a monoclonal cell line capable of stably expressing CD86, 4-1BBL and mbIL-21 proteins is obtained through screening. According to the method, cell lines with the cloning numbers of V5-E9, V10-A5, V5-D6 and V6-D7 are obtained through screening, the four cloning cell lines can serve as feeder cells for stimulating NK cell amplification after being amplified and subjected to irradiation treatment, NK cell amplification can be effectively promoted, the amplification multiple of the NK cells reaches up to 6000-8000, and large-scale preparation and production of the NK cells for treatment are met.

The invention discloses a method for establishing a co-culture system of porcine skeletal muscle satellite cells (PSCs) and porcine precursor fat cells (ADSCs). The method comprises the following steps: selecting a PSCs and ADSCs co-culture proliferation culture medium and a co-culture differentiation culture medium, and establishing an optimal cell co-culture ratio. According to the optimal co-culture proliferation and differentiation culture medium for the PSCs and the ADSCs and the established co-culture proportion, mass proliferation and efficient differentiation of cells can be promoted, the influence of addition of fat cells with different concentrations on muscle cell differentiation can be researched, fat deposition in co-culture is regulated by regulating the proportion of the fat cells, and the differentiation of the muscle cells is promoted. And an earlier-stage foundation is provided for explaining a signal mechanism theoretical basis between cells under growth and development conditions of muscle and adipose tissues and exploring cell culture meat in a later stage.

The invention discloses a rapid screening model for simulating co-culture of colon-like organs and flora, and belongs to the technical field of in-vitro digestion simulation. The invention provides a rapid screening model for co-culture of colon organoid and intestinal flora, and solves the problem that oxygen demands are difficult to coordinate due to different organoid and intestinal flora. By introducing an oxygen gradient control technology, coexistence of organoids in an aerobic environment and intestinal flora in an anaerobic environment is realized in the same system. The rapid screening model disclosed by the invention can efficiently simulate the intestinal microenvironment, is particularly suitable for researching the influence of flora metabolites on the intestinal microecology, and is widely applied to the fields of intestinal health, drug screening, functional food development and the like.

The invention relates to a pulmonary fibrosis model and a construction method and application thereof. The pulmonary fibrosis model provided by the invention is good in stability and controllability and high in maturity, can better simulate functions and structural characteristics of pulmonary alveoli-capillary barriers, has the advantage of high simulation, and has a good application prospect in the aspects of disease simulation, drug detection, screening, evaluation, research and development and the like.

The invention discloses a pig intestine organoid and lymphocyte co-culture system as well as a construction method and application thereof, and belongs to the technical field of virus culture. The co-culture system is obtained by co-culturing porcine ileum organs and porcine small intestine epithelium lymphocytes. By using the co-culture system, the PEDV still can infect the organoid under the condition of not adding pancreatin, and by using the co-culture system to separate and culture the PEDV wild strain, the pathological change of the organoid can be observed during blind passage of the fourth generation, and at the moment, the pathological change does not appear on Vero cells. Reverse transcription-polymerase chain reaction (RT-PCR) identification finds that a stripe can be amplified after a PEDV wild strain is continuously passed for five generations on the co-culture system, and no stripe can be amplified after the PEDV is continuously and blindly passed for five generations on Vero cells no matter whether pancreatin is added or not. The proposal of the invention is of great significance to understanding the pathogenesis of PEDV and developing new drugs and therapies for preventing PEDV infection.

A method for co-cultivation and directed differentiation induction of muscle satellite cells and adipose-derived stem cells is provided. The method includes the steps of co-cultivation and co-differentiation. The method for co-cultivation and directed differentiation induction of muscle satellite cells and adipose-derived stem cells provided by the present disclosure can allow the effective co-cultivation and co-directed differentiation induction of muscle satellite cells and adipose-derived stem cells from Larimichthys crocea, thereby providing a feasible solution for the large-scale production of high-quality cultivated meat.

The invention belongs to the technical field of organ chips, provides an in-vitro brain lymphatic system chip construction method and a brain lymphatic system chip, and makes up the blank of brain lymphatic system in-vitro model construction. The method comprises the following steps: firstly, carrying out flushing treatment, disinfection and sterilization treatment, rinsing treatment and cell culture on a micro-fluidic channel in the chip; then, in the treated micro-fluidic channel, blood vessel channel cell inoculation and brain parenchyma channel cell inoculation are carried out; and finally, in the blood vessel channel, perfusion is carried out once at a preset flow speed every preset duration, shear force stimulation simulating pulse type blood flow is provided, basic formation of in-vitro cerebral vessels is achieved after continuous perfusion, perfusion is carried out once at a preset flow speed every preset duration, shear force stimulation simulating pulse type blood flow is provided, and in-vitro cerebral vessels are formed. Intermittent liquid flow simulating arterial fluctuation is achieved.