125 results about "Cell mass" patented technology

The present disclosure provides a method of purifying or preparing microbial protein concentrate from microbial cell mass. More particularly, the present disclosure provides a method of preparing a microbial protein concentrate, comprising: subjecting a microbial cell mass to a temperature of about 100°C to 210°C and a pressure of about 0.1 bar to 20 bar to obtain a treated microbial cell mass; and separating protein from the treated microbial cell mass to obtain microbial protein concentrate with reduced nucleic acid content and color & odor removed. Additionally, the microbial protein concentrate obtained by the present method has high crude protein content and improved functional properties.

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.

To provide a cell injection device, a cell collection accessory jig, and a cell collection method capable of stably injecting foreign cells into a cell mass regardless of the skill and technique of an operator.SOLUTION: A cell injection device includes: a cell collection accessory jig 24 including a filter section 26 having bubbles of a size that allows a liquid medium D to pass through but not cells Ce; an operation manipulator equipped with an injection pipette 90 capable of collecting and discharging a plurality of cells Ce and the liquid medium D from an opening 90a at the tip; an injection pump capable of adjusting the internal pressure of the injection pipette 90; and a controller that, after collecting a plurality of cells Ce and the liquid medium D inside the injection pipette 90, applies positive pressure to the inside of the injection pipette 90 while pressing the opening 90a at the tip of the injection pipette 90 against the filter section 26, thereby discharging the liquid medium D through the filter section 26 and aligning the exogenous cells Ce inside the injection pipette 90 in rosary-like pattern.SELECTED DRAWING: Figure 2

The invention relates to a cell quantification method and device, computer equipment and a storage medium. The method comprises the following steps: extracting cell contour features of an endoscope cell image corresponding to a region of interest through a pre-constructed deep learning model, and separating a cell contour region corresponding to each cell in the endoscope cell image according to the cell contour features; classifying the cell contour region corresponding to each cell through the deep learning model to obtain a cell category of each cell; determining an overall cell quantification result corresponding to the endoscope cell image through the deep learning model according to the cell contour region of each cell; and performing depth feature extraction on the cell contour region of which the cell category is the abnormal cell through the deep learning model, and determining an abnormal cell quantification result according to the depth feature and the cell contour feature of the abnormal cell. By adopting the method, the abnormal condition of the cells can be accurately and efficiently judged.

The invention relates to the technical field of reproductive biology, in particular to a method for efficiently constructing pig testis organs in vitro. Comprising the following steps: S1, enzymolysis of testicular tissues of piglets; and S2, testicular cell resuspension counting: uniformly suspending the testicular cells obtained after enzymolysis in the step S1, sequentially collecting the cells into a 96-well plate with a U-shaped bottom, and adding a DMEM base culture medium containing 5% of FBS, 10% of KSR and 1% of P / S into each well to complete resuspension culture of the cells. S3, testicular organ in-vitro culture: separating testicular cell masses, transferring the separated cell masses into a 96-well plate with a U-shaped bottom, and adding the cell masses into a DMEM base culture medium containing 5% of FBS, 10% of KSR and 1% of P / S for culture; and the quantity of cells in the 96-well plate with the U-shaped bottom is 5 * 104 cells per well. After the culture system is optimized, the experimental process is simplified, the testicular organs can be directly cultured in the 96-well plate with the U-shaped bottom, the replacement frequency is reduced, the pollution risk is reduced, the number of adopted cells is remarkably reduced, and the culture efficiency is improved.

Hydrogel beads with quantifiable attached biomolecules and their use as cell mimics in cytometric applications are described. The cell mimics described herein are selectively tunable to have at least one optical property substantially similar to the at least one optical property of a target cell (e.g., CD34+ stem cell). The disclosure further relates to methods of using the disclosed cell mimics as enrichment controls in cytometric applications.

The invention relates to a single cell metabonomics-based environmental pollutant toxic effect analysis method, and belongs to the technical field of single cell mass spectrometry. The invention relates to a single-cell metabonomics-based environmental pollutant toxic effect analysis method. The method comprises the following steps: S1, importing mass spectrum flow type original data and extracting a target ion current diagram; s2, identifying a single cell mass spectrum signal and deducting a background signal; s3, matching and identifying metabolites; S4, carrying out data standardization processing and data quality control; S5, identifying differential metabolites; and S6, carrying out pathway enrichment analysis on the screened differential metabolites. According to the analysis method disclosed by the invention, high-quality treatment of single cell metabonomics data is realized, and high-throughput, multi-component and label-free analysis is carried out on pollutants in a single cell.

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 invention provides a multi-organ chip as well as a preparation method and a use method thereof. The multi-organ chip comprises a structural layer and an insulating substrate, wherein the structural layer is provided with a culture chamber, an perfusion channel and a hydrogel perfusion pipeline; a microelectrode array is arranged on the insulating substrate; the limiting part extending from the bottom of the culture chamber to the top of the culture chamber is arranged in the culture chamber, so that the accommodating cavity for accommodating the cell mass or the organoid is formed between the outer side wall of the limiting part and the inner side wall of the culture chamber, and the cell mass or the organoid is limited in the accommodating cavity and is in contact with the hydrogel barrier; due to the affinity between the hydrogel barrier and the cell mass or the organoid, the cell mass or the organoid can be firmly attached to the hydrogel barrier, and the electrode sites of the microelectrodes in the microelectrode array are located at the bottom of the containing cavity, so that the shape and the function of the cell mass or the organoid can be maintained, and the cell mass or the organoid can be prevented from being damaged. And a stable basis is provided for subsequent electrophysiological signal detection.

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 utility model relates to the technical field of culture bottles, and discloses a novel culture bottle suitable for tissue culture, which comprises a culture bottle body, the top of the culture bottle body is fixedly connected with a bottle opening, the interior of the culture bottle body is fixedly connected with a first baffle plate and a second baffle plate, one side of the first baffle plate is an inoculation area, and the other side of the second baffle plate is a liquid changing area. The inner wall of the second baffle is fixedly connected with a filter screen. According to the novel culture bottle suitable for tissue culture, when cells are cultured by tissue blocks, when the tissue blocks are 1-3mm in diameter, the cells climb out more quickly, the quantity of the cells is larger, the tissue blocks with proper sizes can be screened by the novel culture bottle body to enter the culture bottle body for culture, and the cell growth efficiency is improved; when the novel culture bottle body is used for liquid change, passage or cryopreservation, a tissue block can be separated from a culture solution, and a transfer pipette cannot be blocked, so that the operation becomes smoother, the operation efficiency is improved, and the probability of pollution is reduced.

The present invention discloses an automatic focusing method for a single-cell mass spectrometry system based on deep learning. A model is constructed using the Pytorch deep learning framework, including a region filtering mechanism, a feature extraction network, a classifier, and distributed label encoding. A single image is used as the input of the model, and the image is segmented according to actual needs. On this basis, a channel attention mechanism is used to screen the region of interest, filter out background information, and then a high-dimensional semantic feature is obtained using the feature extraction network. These features are passed through the classifier and distributed label encoding to obtain the finally predicted defocus distance, that is, the distance between the current position and the best imaging position. The method of the present invention can greatly improve the real-time performance of the system under the input of high-resolution images.

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 relates to the field of cell analysis related to cytobiology, in particular to a derivatization reagent and application thereof in a method for identifying amino metabolites. The invention provides a novel derivatization reagent 3-bromoquinoline succinimide ester, the derivatization reagent has bromine atoms and quaternary ammonium groups with natural positive charges, so that the response of the labeled metabolite in mass spectrum can be improved, and by utilizing the characteristic of natural isotope distribution of the bromine atoms, the sensitivity of the metabolite in mass spectrum can be improved. The labeled metabolite can generate a pair of isotope peaks with fixed intensity ratio and fixed m / z difference in the mass spectrum, so that the identification of unknown metabolite is facilitated. The living cells labeled by the derivatization reagent can be used for mass spectrometry, especially for the field of single cell mass spectrometry, the response of metabolites in the direct sample injection process is improved, follow-up identification is facilitated, and the blank of domestic related technologies is filled.

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 present application provides methods and systems for improved production of cells (e.g., engineered immune cells) that are readily self-killer. In some cases, the production of engineered immune cells that are readily self-killer expressing chimeric antigen receptor (CAR) molecules that are capable of binding cell surface molecules expressed by other immune cells that likewise express the same CAR molecule may limit the viability and yield of the engineered immune cells. As described herein, methods and systems that allow for control of parameters such as metabolite concentration and cell mass formation during culture may in some cases be used to increase yield and / or viability in the production of cells that are susceptible to self-killer. Also provided herein is an engineered immune cell, a population of engineered immune cells, and / or a pharmaceutical composition comprising an engineered immune cell or a population of engineered immune cells produced using the methods or systems described herein.

Methods are provided for assaying neoplastic cells and / or neoplasia-related cells. Aspects of the methods involve detecting heterogeneity, per cell programmed-death ligand 1 (PD-L1) expression, and / or proliferation of neoplasia and / or neoplasia-related cells. Aspects of the methods include cytometrically assaying a labeled cell suspension to quantify per cell heterogeneity, per cell PD-L1 expression, proliferation, and / or other parameters to detect whether a neoplastic cell is present in the neoplasia sample. Aspects of the provided methods also include treating a subject based on the outcome of such an assay. In addition, kits that find use in practicing the subject methods are also provided.

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 present invention provides a cell sorting device, including a frame, a sorting component and a plurality of clamping mechanisms. The sorting component includes a sorting column and a liquid holding bottle arranged up and down. The open end of the liquid holding bottle can be sealed and connected to the liquid outlet end of the sorting column. The clamping mechanism has a magnetic field, and the clamping mechanism is arranged on the frame. The clamping mechanism is used to clamp the sorting component, and can be kept on the frame or rotated in a vertical plane. The present invention clamps the sorting component by the clamping mechanism to rotate, so that the solution moves back and forth between the sorting column and the liquid holding bottle in the sorting component, so that the target cells in the solution pass through the iron beads in the sorting column for multiple times, thereby increasing the contact between the target cells and the iron beads, thereby increasing the amount of target cells at the sorting location and improving the sorting efficiency.

The present invention provides a fluorinase mutant and a microbial cell displaying fluorinase on the cell surface and its application, belonging to the field of genetic engineering technology. In order to solve the problem of substrate transmembrane restriction in the whole cell catalysis process. The present invention provides a gene with a sequence as shown in SEQ ID NO: 3, and the Escherichia coli engineering bacteria is an Escherichia coli engineering bacteria obtained by using Escherichia coli as a starting strain and expressing an ice nuclease N-terminal encoding gene and a fluorinase encoding gene. The engineering bacteria can stably express the fluorinase gene in Escherichia coli and automatically display it on the cell surface, directly contacting the enzyme with the substrate, avoiding the restriction of the substrate passing through the cell membrane, maximizing the utilization rate of the enzyme, and reducing the influence of exogenous proteins on the host by surface display. A larger cell amount can be obtained, and the possibility of engineering application is improved.

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.

The invention belongs to a mass spectrometry technology, and particularly provides a single cell mass spectrometry detection system and method based on microfluidics, and the single cell mass spectrometry detection system comprises a microfluidic chip and a recognition device; the bearing unit is provided with a plurality of single cell bearing positions distributed in a matrix manner and a waste liquid pool; the driving unit is used for realizing relative movement between the bearing unit and the micro-fluidic chip, so that single cells discharged from the micro-fluidic chip enter a selected bearing position, or liquid without single cells enters the waste liquid pool; the controller controls the driving unit according to a result of the identification device, and the identification device is used for identifying whether a single cell exists in a liquid flow of a single cell channel of the micro-fluidic chip or not. The device has the advantages of controllable droplet size and the like.

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.