10 results about "Intercellular connection" patented technology

The present application relates to the field of image processing, in particular to a kind of dynamic evaluation method and system of endothelial barrier integrity in organ chip.The method obtains the dual-channel fluorescence image of cell nucleus and intercellular junction protein by confocal microscope, and extracts four key feature parameters based on image processing technology: cell coverage, cell nucleus distribution uniformity, intercellular junction protein intensity and junction protein integrity.Through the fusion of these parameters, a comprehensive evaluation index is obtained, and compared with the preset threshold value, to determine whether the endothelial barrier is complete.The present application avoids the interference of traditional permeability detection method, realizes the multidimensional, quantitative and high reliability evaluation of barrier structure, and is suitable for various organ chip models.

The present application relates to the field of aquaculture technology, and particularly relates to a culture method of bigmouth bass gill explants and application. The present application provides a culture method of bigmouth bass gill explants, which uses explant culture solution to culture the gill explants of bigmouth bass. The explant culture solution is formed by adding fetal bovine serum, fish serum, trinity, D-galactose, L-alanyl-L-glutamine, sodium pyruvate and melatonin into a basic culture medium. In combination with dynamic culture conditions, the natural three-dimensional structure and intercellular connection of the gill tissue can be preserved, and the metabolic activity and functional integrity thereof can be maintained. The present application can solve the technical problem that the prior art cannot provide an ideal research model which can both maintain the structural integrity of the gill tissue and accurately reflect the functional characteristics of the gill tissue. Through the culture method, research models such as ammonia nitrogen stress can be efficiently constructed, thereby providing technical support for the research of bigmouth bass gill tissue physiological mechanism, disease prevention and control, environmental risk assessment and the like.

The invention relates to the technical field of aquaculture, in particular to a culture method and application of gill explants of micropterus salmoides. According to the technical scheme, the culture method of the gill explant of the largemouth bass comprises the following steps: culturing the gill explant of the largemouth bass by using an explant culture solution; the explant culture solution is formed by adding fetal calf serum, fish serum, three antibodies, D-galactose, L-alanyl-L-glutamine, sodium pyruvate and melatonin into a basic culture medium. In cooperation with dynamic culture conditions, the natural three-dimensional structure and intercellular connection of the gill tissue can be reserved, and the metabolic activity and functional integrity of the gill tissue can be maintained. According to the technical scheme, the technical problem that an ideal research model capable of maintaining the structural integrity of the gill tissue and accurately reflecting the functional characteristics of the gill tissue cannot be provided in the prior art can be solved. By means of the culture method, research models of ammonia nitrogen stress and the like can be efficiently constructed, and technical support is provided for micropterus salmoides gill tissue physiological mechanism research, disease prevention and control, environmental risk assessment and the like.

The utility model belongs to the field of biotechnology, and particularly relates to a device for preparing a multi-layer cell sheet, which comprises a clamp and a culture box for placing the clamp, the clamp comprises a clamp cover plate and a clamp base. The clamp cover plate is connected to the upper portion of the clamp base. The clamp cover plate is provided with a plurality of cover plate hollowed-out openings, the clamp base is provided with a plurality of base hollowed-out openings, and the cover plate hollowed-out openings and the base hollowed-out openings in the same position are the same in shape. Limiting structures are arranged on the four edges of the clamp base, and the cell sheets are pressed between the limiting structures by the clamp cover plate and then fixed between the clamp cover plate and the clamp base. Compared with the prior art, the positions of the cell sheets in the clamp can be fixed through the cell sheet fixing pieces, the cell sheets are kept in contact in the culture process, then intercellular connection is generated or extracellular matrixes are secreted, and multiple layers of cell sheets are prepared at a time under the condition of not depending on exogenous extracellular matrixes.

The invention relates to the technical field of biology, and discloses a recovery liquid for improving recovery efficiency and vitality of organoids, which comprises a metal ion chelating agent, a dissociation agent and a cell protective agent. According to the recovery liquid, matrigel is mildly disintegrated through cooperation of the metal ion chelating agent and the dissociation agent, the organoid can be separated from the matrix within 30 minutes, and the recovery rate of the organoid reaches 90% or above and is far higher than that of a pure mechanical blowing method; the dissociation agent is used for replacing pancreatin and other strong digestive enzymes, and is matched with the cell protective agent, so that the survival rate of the recovered organoid is greater than or equal to 95%, the three-dimensional structure and the intercellular connection are kept complete, and the biological function is real; all the components are clear, the batch is stable, the operation is simple and convenient, the storage is convenient, the culture medium is suitable for passage, collection, downstream cell counting, molecular detection, immunofluorescence, living cell imaging and the like of various sources of organoids such as gastrointestinal tracts, livers and lungs, and the standardization and repeatability of organoid culture are remarkably improved.

The application discloses a KEGG pathway constrained dynamic graph structure optimization cross-patient cell annotation method, comprising the following steps: obtaining and preprocessing single-cell RNA sequencing data of a reference patient and a query patient; calculating a functional activity score of cells based on a KEGG pathway classification system, and constructing a KEGG function semantic matrix; constructing an initial sparse kNN cell graph; using the KEGG function semantic vector as a biological constraint, dynamically adjusting the connection weight between cells through a multi-view gated similarity calculation and an iterative optimization mechanism, and generating a patient-specific cell graph containing function semantic information; constructing a shared graph encoder and a classifier, and introducing a Wasserstein discriminator for adversarial training to realize cross-domain alignment of embedding distribution of the reference patient and the query patient; and finally predicting the type of the query patient cell by using the trained model. The application solves the problem of significantly improving the accuracy, robustness and biological interpretability of cross-patient cell annotation.

This application discloses an in vitro reconstructed vaginal mucosa model, its construction method, and its application. The construction method includes an improved culture medium: HVU medium is added to immortalized vaginal epithelial cell line VK2 / E6E7, which is then seeded into Transwell chambers and cultured. The HVU medium on the surface of the culture in the Transwell chambers is aspirated, and the Transwell chambers are raised to the air-liquid interface for air-liquid surface culture. The culture medium is then replaced with HVA medium to obtain the vaginal mucosa model. This method improves the stability of cells during in vitro culture and constructs an in vitro reconstructed vaginal mucosa model with a tissue structure highly similar to human vaginal mucosa (possessing a complete stratified squamous epithelial structure and normal intercellular connections), thereby enhancing the model's physiological functional simulation.

The invention provides an enzymatic self-assembly polypeptide, an NK cell modified by the enzymatic self-assembly polypeptide and a preparation method and application of the NK cell, and belongs to the technical field of tumor immunotherapy. The preparation method comprises the following steps: incubating NK cells and Ac4ManNAz to obtain NK-N3 cells; the NK-tumor cell connection enhancer NBD-GFFpYK (DBCO) RGD is sequentially mixed with the cancer cells and the NK-N3 cells for incubation, or the NK-tumor cell connection enhancer NBD-GFFpYK (DBCO) RGD is sequentially mixed with the NK-N3 cells and the cancer cells for incubation. According to the invention, the adhesion efficiency and targeted killing ability of NK cells and cancer cells can be significantly enhanced, and by means of a dual-targeting mechanism of enzyme response and a membrane receptor, it is ensured that the NK-tumor cell connection enhancer is only enriched at a tumor site, and the influence on normal cells is reduced; a nanofiber network formed by the self-assembled polypeptide can resist tumor microenvironment interference and stably maintain intercellular connection.

The invention relates to the field of image processing, in particular to a dynamic evaluation method and system for the integrity of an endothelial barrier of an organ chip. According to the method, a two-channel fluorescence image of the cell nucleus and the intercellular connexin is obtained through a confocal microscope, and four key characteristic parameters including the cell coverage rate, the cell nucleus distribution uniformity, the intercellular connexin strength and the connexin integrity are extracted on the basis of an image processing technology. A comprehensive evaluation index is obtained by fusing the parameters and is compared with a preset threshold value, so that whether the endothelial barrier is complete or not is judged. According to the method, interference of a traditional permeability detection method is avoided, multi-dimensional, quantitative and high-reliability evaluation of the barrier structure is achieved, and the method is suitable for various organ chip models.

The application belongs to the technical field of cell engineering and cell differentiation regulation, and discloses a method for directionally inducing secretion function cells from iPSCs and application thereof. The method takes the initial state of iPSCs and the potential of secretion function as cores, and constructs a pretreatment-induction differentiation integrated system. By screening and passing, iPSC clones with specific adhesion characteristics and enzyme sensitivity are enriched, and an iPSC population with enhanced secretion function potential is obtained. In the differentiation initiation stage, a culture system containing a Rock inhibitor is used in combination with laminin modified culture surface to maintain the intercellular connection state and realize the synchronous response of the induction signal. Then, the directionally induced differentiation is carried out by adding induction factors in stages, and the obtained cells are screened and purified to prepare high-purity secretion function cells. The prepared cells can be widely applied to drug screening, cell therapy and disease model construction, and provide technical support for the large-scale preparation of clinical-grade secretion function cells.