43 results about "Cell mass" patented technology

The invention belongs to the technical field of medical diagnosis, and discloses a quantum multicolor immune digital pathological diagnosis analysis system, which comprises the following steps: acquiring high-resolution digital images of an immunohistochemical or immunofluorescence slice and a slide, and analyzing multicolor signal distribution in the images to obtain a quantum chromatographic image set; pixel-level color separation and intensity quantization are carried out on the quantum chromatographic image set, and a quantum intensity index set of each pixel is generated; performing multi-target color overlay analysis and post-translational modification protein specificity evaluation on the quantum intensity index set to construct a multi-color modification index set; identifying a single cell boundary and a multi-cell community based on the multicolor modification index set, and extracting a cell morphological parameter and a spatial adjacency relation to obtain a cell quantum analysis set; performing consistency correction and traceable recording by applying an automatic quality control mechanism, and generating a digital diagnosis report; the accuracy of accurate qualitative and quantitative analysis of immunohistochemistry and immunofluorescence is greatly improved.

The application discloses a kind of artificial cells for single-cell mass spectrometry and preparation method thereof, wherein artificial cell includes internal water phase, intermediate oil phase and external water phase;Internal water phase includes polyethylene glycol and polyvinyl alcohol aqueous solution, intermediate oil phase includes L-alpha-phosphatidylcholine chloroform and hexane mixture;External water phase includes PVA and F-68 aqueous solution.Compared with natural cell sample, the novel artificial single cell based on microfluidic self-assembly developed in the application has better uniformity, stability and controllability, effectively avoids significant measurement difference between single-cell individual samples, and effectively solves the problem of difficult stable preservation of biological samples.The proposed artificial cell preparation method will largely solve the problem of lack of standard reference material in the field of single-cell mass spectrometry, making the results of single-cell mass spectrometry method more accurate, reliable and mutually recognized.

The purpose of the present invention is to three-dimensionally culture osteoblasts produced with a direct conversion technique, and to provide a three-dimensional bone-like tissue composed of the osteoblasts and bone matrix. The present invention relates to, for example, a method for producing an osteoblast-containing three-dimensional cell mass, comprising: a first step for performing direct conversion from somatic cells of mammals into osteoblasts; a second step for subjecting cells having undergone the first step to high-density culture, and forming a cell sheet containing the osteoblasts; and a third step for obtaining an osteoblast-containing three-dimensional cell mass from the cell sheet obtained in the second step.

The application discloses a method for inducing dedifferentiation of anther of Lagerstroemia indica to form a callus. The application selects the anther of Lagerstroemia indica as an explant for dedifferentiation induction, and through a large number of experiments, the most suitable dedifferentiation culture medium formula and culture conditions are screened out, thereby providing reference value for future development of related tests. The explant inoculation contamination rate is less than 10%, the average dedifferentiation culture rate can reach 35%, and the callus cell mass develops healthily. The dedifferentiation induction efficiency of the woody Lagerstroemia indica anther is improved.

The invention provides a method for producing platelets through human iPSCs (induced pluripotent stem cells) in-vitro differentiation, aiming at the existing in-vitro induction method, a systematically optimized in-vitro culture scheme is adopted, and the method mainly comprises the following four aspects: (1) optimizing forming conditions of embryoid bodies (EB); (2) optimizing the composition of a basic cell culture medium; (3) establishing an induction scheme for replacing cell factors with small molecular compounds; and (4) adding a small-molecule inhibitor to promote maturation and differentiation of megakaryocytes. The efficiency is improved by a technology for promoting MKs maturation by adjusting the amount of initial EB cells and adding small molecules, the cost is reduced by a method for optimizing culture medium components and replacing cell factors with small molecules, and the differentiation time of megakaryocytes is shortened to 14 days; the differentiation efficiency of the MKs is remarkably improved by about 2.5 times (about 1.428 megakaryocytes are obtained by differentiating each iPSC, and about 0.57 megakaryocytes are obtained by differentiating each iPSC according to a basic scheme), and the final differentiation maturity is remarkably improved. Meanwhile, after the cell culture medium is optimized, the production cost is obviously reduced by 70.9%.

The invention discloses a pig jejunum crypt separation and organoid model construction method, and belongs to the technical field of animal tissue separation and culture. The method specifically comprises the following steps: intestinal tract collection, cleaning solution preparation, intestinal tract treatment, tissue trimming, tissue cleaning, digestive juice preparation, tissue digestion, cell mass separation, crypt separation, crypt counting, crypt collection, crypt inoculation and organoid culture. The porcine jejunum organ obtained by the method has a typical three-dimensional cystic structure, has complete epithelial polar and functional cell types, including microvillus, goblet cells, panning cells and endocrine cells, and shows good barrier function and drug metabolic activity. A reliable in-vitro model platform is provided for pig intestinal development, nutrient absorption, pathogen infection mechanism research and precision medical treatment, meanwhile, a technical foundation is laid for large-scale application of organoids, and the method has important scientific significance and clinical value.

The invention provides a macrophage cell burial function quantitative analysis and evaluation method based on image processing, and relates to the technical field of image processing, and the method comprises the following steps: obtaining cell quantification data and space transcriptome data of a biological sample; the cell quantitative data comprises quantitative kinetic parameters of a cell burial process and a quantitative protein expression level; the spatial transcriptome data comprises gene expression information and spatial position data thereof; based on a graph neural network model, carrying out fusion analysis on the cell quantification data and the spatial transcriptome data, taking the identified single cell as a graph node, and taking the spatial proximity relationship between the cells as a graph edge; by analyzing the connection relation between the attribute features of the graph nodes and the graph edges, the spatial distribution mode of the cell burial events is quantified, the cell ecological niche with active cell burial is identified, and the comprehensive index for comprehensively evaluating the cell burial function of the macrophages is calculated. The invention provides a brand-new and powerful analysis tool for functional analysis of the immune microenvironment.

This disclosure relates to methods of washing cells from a grown cell mass to remove cell culture media and enriching the cells with a finishing media. The disclosed method includes growing a cell mass in cell culture media and then collecting and washing the grown cell mass with a series of wash buffers or a gradient wash buffer that changes over time. While the cell culture media contains nutrients and components beneficial for cell growth, cells grown in cell culture media often have off flavors, off aromas, poor color, poor salt / minerality compositions, and other shortcomings. Accordingly, the disclosed method comprises removing cell culture media remnants from a grown cell mass using a single or a series of wash media. The grown cell mass is further washed with a finishing or enrichment buffer to further improve sensory aspects and nutritional composition of the grown cell mass.

According to one embodiment of the present invention, provided are: a cell mass-forming member that is capable of easily forming a cell mass and superior in industrial mass productivity; a culture container equipped with the cell mass-forming member; a method for producing cultured cells using the cell mass-forming member; and cultured cells with a cell mass-forming member that are equipped with the cell mass-forming member. The cell mass-forming member 1B according to one example of the present invention has a base material 2, wherein: an adhesion inhibition area 3A and a cell adhesion area 4B are formed on the surface 2a of the base material 2; a micro-concavo-convex structure area 6 including a plurality of convex portions 7 is formed in the cell adhesion area 4B; and a hydrophilic coating layer 5 is formed on both the adhesion inhibition area 3A and the cell adhesion area 4B. The culture container and the cultured cells with a cell mass-forming member according to one example of the present invention are equipped with the cell mass-forming member 1B. The method for producing cultured cells according to one example of the present invention comprises using the cell mass-forming member 1B.

The invention belongs to the technical field of single-cell metabolome analysis, and particularly relates to a single-cell metabolome analysis method and system with a motility rate sensing function, and the method comprises the steps: obtaining metabolism spectrum data of a plurality of single cells through a single-cell mass spectrum platform, and forming an initial single-cell metabolism matrix; the initial single cell metabolism matrix comprises a plurality of single cell samples and corresponding metabolite ion strength characteristics; inputting single cell survival rate labels provided by an external survival rate judgment module into the analysis process, wherein the survival rate labels are in one-to-one correspondence with corresponding single cells of the initial metabolism matrix; performing subset division on the initial metabolism matrix according to the motility rate label input in the step 2; reconstructing a living cell metabolism matrix and a dead cell metabolism matrix in groups, performing data processing and analysis operation on the living cell metabolism matrix and the dead cell metabolism matrix, and outputting an analysis result with better biological interpretation based on motility rate correction.

The utility model relates to the field of biomedical technology, and disclose multi -passage liquid -based cell collection and separation filter kit;Including box body and connecting pipe, the outside of connecting pipe is fixed with support frame, the inside of support frame is provided with dismounting assembly, the one side of dismounting assembly is provided with clamping ring, the one end of connecting pipe is linked with collecting pipe;The utility model discloses through pressing multiple groups of pressing type hose simultaneously, drives the collection head to be through sample pipe and make sample be drained to the flow guide pipe through the flow guide type hose quickly, then imports the filter pipe from the flow guide pipe, realizes multi -passage parallel collection, greatly promoted sample collection efficiency, after sample enters filter pipe, passes through rough filter screen, middle filter screen and fine filter screen in proper order, and rough filter screen effectively intercepts the large particle impurity in sample, and middle filter screen further filters cell mass, and fine filter screen then accurate capture target cell, reach the fractional separation of cell, ensure the accuracy and reliability of subsequent detection.

The invention discloses a sensing chip and a detection method thereof. The invention relates to a sensing chip, which comprises a sensing substrate provided with a plurality of optical waveguide detection units; each optical waveguide detection unit corresponds to one cell culture bin, and a polydimethylsiloxane sheet covering the optical waveguide detection unit is arranged in the middle area in each cell culture bin; the optical waveguide detection unit comprises an input waveguide, an annular waveguide and an output waveguide, and the polydimethylsiloxane sheet is arranged above the optical waveguide detection unit in a covering manner. The invention discloses a method adopting an optical waveguide as a core detection unit, which is high in flux, high in speed, small in required cell quantity and greatly reduced in manual intervention part operation, and provides a rapid and reliable method for cell migration detection.

The invention belongs to the technical field of cell viability on-line discrimination, and particularly relates to a mass spectrum-based cell viability measurement method and a mass spectrum-based cell viability measurement system. The invention provides a label-free endogenous metabolite-based cell viability measurement scheme and a label-free endogenous metabolite-based cell viability measurement system, aiming at the defects that the prior art depends on an exogenous label and cannot be synchronously carried out on line with single cell mass spectrometry. The method comprises the following steps: acquiring continuous single cell metabolome data through single cell electrospray ionization mass spectrometry, identifying a single cell event and a time window by taking ubiquitous membrane lipid as a first characteristic ion, extracting an endogenous metabolite reflecting cell activity in the window as a second characteristic ion, and calculating a signal-to-noise ratio; generating an activity label according to a preset threshold value and counting the cell viability of the group, wherein the system correspondingly comprises a data acquisition module, an event identification module, a marker extraction module, a threshold value judgment module and a viability counting module. According to the invention, synchronous, on-line and label-free accurate measurement of the cell viability in single cell metabonomics analysis is realized, and exogenous interference and data deviation are effectively avoided.

The utility model provides a kind of microfluidic chip, including the mixing layer, dilution layer, filter layer and focusing layer that are sequentially stacked, each functional layer internal flow passage sequence communication, to build a compact structure, complete automation cell sample processing system of function.Firstly, mixed flow channel is used to mix cell liquid and diluent, then through mixing and diluting cavity, cell liquid is efficiently and uniformly gradient diluted;Subsequently, sample flow is filtered to accurately remove impurities and cell mass, improve the purity of sample.Finally, the liquid focusing section in focusing layer can arrange the processed cells by hydrodynamics, so that it is arranged in order to pass, to create ideal conditions for subsequent cell counting device accurate optical detection and counting.By sequentially stacking the four functional layers of mixing, dilution, filtration and focusing, the overall volume of the microfluidic device is significantly reduced, and the automation and optimization of the whole sample processing process are realized.

The present invention provides a system for performing high-throughput selection of cell masses from a source, fabrication of biomaterial-encapsulated cell mass model based on the selected cell masses, and separation of biomaterial-encapsulated cell mass model, cell masses or cells from a pool of biomaterial-encapsulated cell mass models as-fabricated in a fully automated manner to minimize selection errors due to human intervention and potential contaminations to biological samples during liquid handling among different analytical devices or units.

The invention provides a tumor drug evaluation method based on PBMC (peripheral blood mononuclear cells) and prostate organs and application, and belongs to the technical field of biology. The method comprises the following steps: (1) constructing a tumor organoid, namely pretreating an excision tissue to obtain a cell cluster, wrapping the cell cluster by using Matrigel, paving a plate, and adding a tumor organoid culture medium for culturing; (2) separating peripheral blood mononuclear cells; (3) co-culturing the peripheral blood mononuclear cells and the tumor organs to obtain CTL cells; and (4) co-culturing the CTL cells and the tumor organoid, treating the co-cultured mixture with the tumor drug, and judging the efficacy of the tumor drug. According to the tumor drug evaluation method based on the PBMC and the prostate organoid, the tumor microenvironment can be relatively closely reproduced by utilizing the tumor organoid, and the efficacy of the anti-cancer drug can be more accurately evaluated.

The invention provides an ELISPOT (Enzyme-Linked Immunospot Assay) analysis method for detecting IFN-gamma and IL-2 secretion, which comprises the following steps: activating an ELISPOT plate, washing, adding IFN-gamma and IL-2 capture antibody coating with specific concentration, and staying overnight at 4 DEG C; recovering and counting target sample cells, resuspending the target sample cells according to a certain concentration, and adding the target sample cells into coated plate holes; incubating for more than 16 hours in an environment of 37 DEG C and 5% COC; washing the incubated plate, adding IFN-HRP and IL-2-ALP detection antibodies, and incubating at room temperature for 2 hours; after washing, adding a blue developing solution for developing for 10-15 minutes; adding a red developing solution, incubating for 10-15 minutes, and stopping developing; according to the method, T cell subset identification and multifunctional analysis can be carried out, in the early stage of vaccine research and development, due to the fact that target positive population is small, the number of obtained PBMC cells is small, two-factor detection can detect different cell factor reaction frequencies and locate different cell populations and subtypes at the same time, and comparison of two-factor data is beneficial to finding out population immune differences; and scientific research and vaccine development are promoted.

Provided herein are texturization methods that allow for replication of the fibrous texture of animal flesh using a large amount of cells while using a high throughput methodology at temperatures below typical extrusion temperatures. The texturization methods incorporate Plantago seed-derived polysaccharide, including for example, psyllium. Suitable sources of cells may be used in such texturization, including animal-derived cells, finfish-derived cells, and yeast cells. The resulting texturized cell mass has visual fibrosity, and may be used to produce one or more downstream products, including food products.

The present application relates to the field of biotechnology, in particular to human bladder tissue digestion solution and application thereof.The digestion method of the present application can dissociate human bladder tissue cells while keeping the cell viability at more than 90%, and the amount of harvested cells is also obviously improved.Sufficient cell amount is the prerequisite for optimization treatment such as red blood cell lysis and dead cell removal.The single cell suspension obtained by the method has high viability, low clumping rate, high nucleated rate and less impurities, which can provide a basis for obtaining high-quality gene expression data in human bladder tissue single cell sequencing experiment.The present application can quickly obtain human bladder tissue primary cell suspension (sufficient cell amount, high viability, low clumping rate and less impurities) meeting the requirements of single cell experiment within 2-4 hours.

The present invention provides a method for producing an artificial bone-cartilage complex, the method comprising: (i) a step for inducing differentiation of pluripotent stem cells to obtain a precartilage cell mass; (ii) a step for seeding and culturing the precartilage cell mass obtained in the step (i) on the surface of an artificial bone having communication holes; and (iii) a step for recovering an artificial bone-cartilage complex in which a cartilage layer is formed on the surface of the artificial bone. Also provided is an artificial bone-cartilage complex produced by said method.

The invention discloses an application of methyl cellulose in suspension culture of pancreatic cell masses. The invention also provides a method for suspension culture of a pancreatic cell mass, a pancreatic cell mass culture produced by the method, and a pharmaceutical composition and application of the pancreatic cell mass culture. The method provided by the invention comprises the step of carrying out suspension culture on pancreatic cell clusters in a culture medium containing methylcellulose, the number of the cell clusters cultured and harvested by the method is more, the average diameter of the cell clusters is better controlled, and the expression proportion of related markers is higher; the method has a good application prospect in preparation of regenerated pancreas islet tissues and treatment of diseases or symptoms related to pancreas islet function impairment.

The invention provides a universal tissue single-cell suspension preparation kit and a use method thereof. The kit provides six common tissue dissociation enzymes and auxiliary materials required by the whole process of single-cell suspension preparation such as dissociation, cleaning, red blood cell lysis, fragment removal and cell mass removal, and can be used for rapidly preparing single-cell suspensions for at least 12 tissues (including difficult tissues such as skin, muscle, fat, pancreas and intestines). The components are complete, extra reagents are not needed when the single-cell suspension is applied to a conventional scene of single-cell suspension preparation, and the compatibility to personnel and a platform system is good. According to the kit, domestic raw materials are used, the cost is only one fifth of that of imported raw materials, and the single-cell suspension prepared by the kit is high in cell yield, good in activity, few in impurities, low in caking rate and high in RNA integrity, so that high-quality single cells can be conveniently and rapidly obtained for downstream capture and analysis; therefore, data waste and gene transcriptome expression information distortion caused by poor cell state or RNA degradation are reduced.

The application discloses a centrifugal tube and assembly for making cell wax blocks. The centrifugal tube comprises a ball-sac-equipped inner thread top cover, a centrifugal tube body composed of a funnel part and a cell enrichment tube, a cell collection capsule, a bottom pad and a bottom-sealed inner thread cover. The cell collection capsule and the bottom pad are arranged inside the cell enrichment tube to collect cells and can be taken in and out of the lower opening of the cell enrichment tube. After installation, detection liquid is added to the centrifugal tube body, and the centrifugal tube body is centrifuged by a centrifugal machine. The cells in the liquid are settled in the collection capsule. The bottom thread cover is unscrewed, the ball sac is pressed, and the cell collection capsule with the internal cell sediment is discharged from the lower opening of the cell enrichment tube under the hydraulic action. Then, the cell collection capsule is taken with the capsule cap and is placed in 10% neutral formalin solution for fixation. The fixing solution gradually penetrates into the cell mass, and the cell sediment gradually forms a certain shape and hardness. In this way, the cell wax block is simple and convenient to make and is not limited by the small amount of cells.

The application provides a biomarker for diagnosing GVHD and application thereof. The application proves by experiments that the Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in donor-derived T cells can be used as biomarkers for distinguishing GVHD patients from non-GVHD patients, and when the T cells cannot be distinguished from the donor and the recipient, the Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in the T cells can still be used as biomarkers for distinguishing GVHD patients from non-GVHD patients. The method for diagnosing GVHD by using the biomarker provided by the application has the advantages of immune subpopulation specific quantification, low cell amount adaptability, short time efficiency, low cost and the like.

The invention discloses a suspension cell continuous culture process and an impurity removal method of a cell suspension, and belongs to the technical field of cell culture. In the continuous culture process of the suspension cells, dead cells, cell agglomerates and / or cell debris are removed by adjusting the standing and settling conditions, especially the temperature, so that damage to the cells caused by centrifugation, introduction of exogenous reagents and the like is avoided, and the high cell viability is maintained. Dead cells, cell agglomerates and / or cell debris in a cell suspension in a reactor are regularly removed in a continuous culture process, so that single cells with high cell activity and high purity are continuously obtained, rapid and high-value multiplication of the cells is realized, and a guarantee is provided for the stability of subsequent experimental research and industrial production.

Methods that include tagging at least one cell with a MXene, the cell optionally being an immune cell; and detecting at least one component of the MXene using one or more of single-cell mass cytometry by time-of-flight (CyTOF), imaging mass cytometry (IMC), and ion beam imaging (MIBI-TOF). Systems that include a cell tagged with an amount of a MXene; and a detection train configured for at least one of time of flight (CyTOF), imaging mass cytometry (IMC), and ion beam imaging by time-of-flight (MIBI-TOF) that detects the MXene. Methods that include tagging a population of cells with at least one MXene: and processing the population of cells with at least one of time-of-flight (CyTOF), imaging mass cytometry (IMC), and ion beam imaging by time-of-flight (MIBI-TOF) that detects the at least MXene; and relating the detection of the at least one MXene to a characteristic of the population of cells.

The application discloses a single-cell mixed sample sequencing and sample splitting method based on HLA genes. The application first acquires HLA gene typing information of each sample to be mixed; secondly, single-cell mixed sample sequencing is performed on the mixed sample to obtain single-cell mixed sample sequencing results of the mixed sample; finally, based on the HLA gene typing information, the sample source of each cell is identified by calculating the expression characteristics of HLA genes in the single-cell mixed sample sequencing sequence, and sample splitting is completed. The application breaks through the technical obstacle that few-cell clinical samples cannot be subjected to single-cell sequencing, greatly improves the utilization rate of these precious samples, significantly reduces the calculation consumption of sample splitting through an efficient HLA gene sequence pseudo-alignment algorithm, improves the ability to obtain genetic information from low-coverage cells by focusing on the HLA region with the highest polymorphism in the human genome, and effectively reduces the sequencing cost of a single sample through mixed sample sequencing.

The invention relates to the technical field of biotechnology, and discloses a construction method of a bovine originating embryonic stem cell line, which comprises the following steps: (1) obtaining in vitro fertilized bovine early blastocyst; (2) trophoblast cells of the blastocyst are removed, an inner cell cluster is inoculated into a culture system containing a feeder layer, and the culture system is a basic culture medium added with FGF2 and WNT pathway inhibitors; and (3) culturing until an inner cell cluster forms a cell set drop, and carrying out digestion passage to obtain the stably proliferated bovine originated embryonic stem cell line. According to the construction method and the application of the bovine embryonic stem cell line, a specific initial induction combination of 'FGF2 + WNT inhibitor IWR1' is adopted, and a cell mass in an early blastocyst is used for establishing the line, so that the bovine embryonic stem cell line which is uniform in form, can be stably passaged for more than 26 generations and still keeps typical embryonic characteristics is successfully obtained; the problems of unstable state and easy differentiation of the bovine stem cell line in the prior art are solved.

The present invention aims to provide a method for efficiently producing a cell mass containing pituitary tissue from pluripotent stem cells. A method for producing a cell mass containing pituitary tissue, including the following steps (1) and (2):(1) a first step of suspension-culturing pluripotent stem cells to form a cell aggregate in the presence of a Wnt signal transduction pathway inhibiting substance,(2) a second step of suspension-culturing the aggregate obtained in the first step in the presence of a BMP signal transduction pathway activating substance and a Sonic hedgehog signal transduction pathway activating substance, thereby obtaining a cell mass comprising pituitary tissue.