67 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 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.

The utility model discloses a cell transfer bag with a novel structure, which comprises a transfer bag which is respectively connected with an input port, a liquid inlet conduit and an output port, and is characterized by further comprising a sampling bag which is communicated with the transfer bag through a plastic hose and is a closed bag body. The cell preparation sampling device is novel in structure and high in practicability, the upper end of the transfer bag is connected with the sampling bag through the plastic hose, when a cell preparation needs to be sampled, 1-2 ml of cell suspension is manually squeezed into the sampling bag, then the plastic hose between the sampling bag and the transfer bag is subjected to heat sealing, after heat sealing is completed, the sampling bag is taken down, and then cell activity and cell quantity detection can be conducted; the sampling is convenient and sterile.

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 belongs to the technical field of gene engineering, and particularly relates to a CHO cell transient transfection method and application thereof.The CHO cell transient transfection method comprises the following steps that subcultured CHO cells are resuspended through a transfection culture medium, the cell density in CHO cell suspension is 0.8 * 10 < 7 >-1.5 * 10 < 7 > cells / mL, a PEI transfection reagent containing 40 kDa linear PEI is used, plasmid DNA and the PEI transfection reagent with the mass ratio being 1: (1-6) are added into the CHO cell suspension for transfection, and transfection is conducted after the PEI transfection reagent is added into the CHO cell suspension; the addition amount of plasmid DNA in each milliliter of CHO cell suspension is 3.0-7.0 [mu] g, transfecting for 20-24 h, then adding a cell suspension culture medium, transfecting for 48-72 h, then repeatedly transfecting, and culturing to obtain the recombinant CHO cell. According to the method, the transfection efficiency can be improved, more cells for expressing the recombinant protein can be obtained, and the transient expression of the target protein can be remarkably 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 present application relates to the field of cell analysis related to cell biology, in particular to a kind of derivatization reagent and its application in the method for identifying amino metabolites.The present application provides a new derivatization reagent 3-bromoquinoline succinimidyl ester, the derivatization reagent has bromine atom and quaternary ammonium group with natural positive charge, which can improve the response of labeled metabolites in mass spectrometry, and using the characteristics of natural isotopic distribution of bromine atom, the labeled metabolites can produce a pair of isotopic peaks with fixed intensity ratio and fixed m / z difference in mass spectrometry, thereby facilitating the identification of unknown metabolites.The living cells labeled by the derivatization reagent of the present application can be used for mass spectrometry, especially in the field of single-cell mass spectrometry, to improve the response of metabolites in direct injection process and facilitate subsequent identification, and make up for the blank of domestic related technology.

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 biomarker for diagnosing GVHD and application of the biomarker. Experiments prove that 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 and non-GVHD patients, and when donor-derived T cells cannot be distinguished, Ki67 positive proportion, Granzyme B positive proportion and TEM cell proportion in T cells can still be used as biomarkers for distinguishing GVHD patients and non-GVHD patients. The method for diagnosing GVHD by using the biomarker provided by the invention has the advantages of immune subgroup specific quantification capability, low cell quantity adaptability, short timeliness, low cost and the like.

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.

A cell culture tool configured to be produced in a simple facility to obtain a cell mass having a desired shape. The cell culture tool includes a cell culture base layer that contains a cell culture base. The cell culture base layer has a cell adhesive area to which a cell is adherable and a cell adhesion inhibitory area where cell adhesion is inhibited. The cell adhesion inhibitory area includes a modified product of the cell culture base.

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 discloses a method for detecting a Br element in a single cell based on a mass spectrum flow type. The detection method comprises the following steps: collecting a biological sample exposed by a brominated compound; preparing a single-cell suspension from a biological sample; labeling cells in the single-cell suspension with metal isotopes A and B respectively; the metal isotope A can be used for simultaneously marking survival and dead cells; the metal isotope B can mark dead cells; and performing single cell mass spectrometry flow detection on the single cell suspension, firstly separating single cells according to the signal of the metal isotope A, recording the single cells as a set S, then obtaining individuals with the signal of A but not with the signal of B from the set S, recording the individuals as a set P, and obtaining the single cell mass spectrometry flow detection result through a standard curve according to the detected signal intensity value of the Br element in the cells. According to the method, the Br content in a single cell can be detected, a single-cell inductively coupled plasma time-of-flight mass spectrometer is used for detection, the flux is high, and the analysis speed can reach 600 cells per second.

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.

The invention discloses a mass spectrum image super-resolution reconstruction method and device for a single cell, and the method comprises the steps: carrying out the morphology representation of the single cell through an optical microscope, obtaining a single cell optical image, and obtaining a single cell low-resolution mass spectrum image through a mass spectrum imaging experiment; then registering the single-cell optical image and the single-cell low-resolution mass spectrum image by using a registration module; in order to prevent optical diffraction interference, filtering the registered single-cell optical image through guided filtering, and inputting the registered and filtered single-cell optical image and a single-cell mass spectrum imaging graph into a multi-modal fusion module to generate a mass spectrum super-resolution reconstruction image; and finally, in order to be closer to an image imaged by a real mass spectrum instrument, a process of imaging simulation to the real instrument is carried out, and after simulation, the super-resolution image can be closer to a real mass spectrum high-resolution image. According to the invention, high-spatial-resolution mass spectrum imaging can be realized by using the existing low-resolution mass spectrometer.

The invention provides a method for genetically modifying a cell by using a lambda Red recombination technology, which comprises the following steps: (a) preparing a cell to be modified, namely culturing the cell with a lambda Red recombination system at a first temperature until OD600 is equal to 0.1-0.2; inducing expression of a lambda Red recombination system in the cells, and culturing the cells at a second temperature until OD600 = 0.4-1.5, the second temperature being at least 4 DEG C higher than the first temperature; cooling and centrifuging the cells to obtain a cell block mass; resuspending the cell mass with a glycerin solution to obtain a to-be-modified cell suspension; (b) preparing a targeting fragment, comprising: linearizing a plasmid comprising an exogenous DNA fragment; amplifying the exogenous DNA fragment by taking the linearized plasmid as a template; and recovering an amplification product as a targeting fragment, wherein the targeting fragment comprises the exogenous DNA fragment and homologous arms positioned on two sides of the exogenous DNA fragment; and (c) introducing the targeting fragment obtained in the step (b) into the to-be-modified cell obtained in the step (a), and continuously culturing the cell at the temperature of not less than 35 DEG C.

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 invention discloses a cylindrical cell mass spectrum detection cavity which comprises a bearing assembly, a middle air supply structure and an edge air supply structure, the bearing assembly is driven by the lifting mechanism to define a vacuum cavity for placing the detection frame with the sealing cover, and the bearing plate is used for placing the detection frame; the middle air supply structure comprises a first small hole communicated with the vacuum cavity and located in the middle of the bearing plate, a first sealing plate covers, seals and fixes the first small hole from bottom to top, a first cavity is formed between the first sealing plate and the first small hole, and the first sealing plate is provided with an air supply connector communicated with the first cavity; the edge air supply structure comprises a plurality of second small holes which are communicated with the vacuum cavity and located in the lower edge of the bearing plate, the edge sealing plate covers, seals and fixes the second small holes from bottom to top, and a second cavity is formed by the edge sealing plate and the bearing plate; the edge sealing plate and the first sealing plate are respectively provided with a first joint and a second joint which are connected through a pipeline. The structure enables the middle and peripheral areas of the detection frame to be communicated, the detection effect of a mass spectrum detection instrument is improved, and large-scale detection of cells can be efficiently realized.

In a cell culture plan creation device (40), an input unit (142) receives an input of a cell culture completion period and a target cell amount at the culture completion period, and a plan creation unit (144) creates a cell culture plan. The cell culture plan includes a first execution designation for one or more subculture processes sequentially executed so as to obtain a target cell amount at a culture end period, and a second execution designation for a culture process executed subsequent to each subculture process. The first execution designation includes information that specifies a type of the passage process and information that specifies a period of execution of the passage process.