79 results about "Cell separation" patented technology

The invention provides a precise magnetic control whole blood rare cell flow magnetic separation system, a magnetic separation and identification method and application, and belongs to the technical field of micro-control flow biological detection. According to the invention, the magnetic separation chip can form a gradient magnetic field, and cells are fixed at a certain position on the sealing film under the combined action of magnetic field force and friction force, so that the separation of the cells is realized. Whole blood cell detection is carried out based on the gradient magnetic field, whole blood sample loading treatment can be directly carried out, the problem that the capture rate is reduced due to mechanical death, damage and stress gene expression caused by repeated elution / transfer steps is avoided, and the capture efficiency and cell activity of the chip are guaranteed; the magnetic field provided by electricity is not needed, the cost is low, the detection accuracy is high, the problems existing in existing CTC detection are solved, economic and practical values are achieved, further development of CTC detection can be promoted, and a novel and effective method is provided for complete blood cell separation detection.

The invention relates to a method for detecting activation of specific T cells after tuberculosis infection and application of the method. Specifically, the invention provides a reagent combination for detection, and the reagent combination comprises an anti-CD4 antibody (CD4 antibody), an anti-CD154 antibody (CD154 antibody), an IFN-gamma detection antibody and a tuberculosis specific antigen. According to the method, through peripheral whole blood collection, PBMC cell separation, T cell activation (activation), activation termination (blocking), antibody labeling, membrane rupture, fixation, intracellular cytokine dyeing, flow cytometry detection and result analysis and judgment, the ratio of the expression IFN-gamma cells in the CD4 + CD154 + cells can be rapidly and accurately determined, and by combining an ROC curve and a constructed judgment standard, the content of the IFN-gamma cells in the CD4 + CD154 + cells can be determined, and the content of the IFN-gamma cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells in the CD4 + CD154 + cells can be determined. And the sample to be detected is judged to be in a positive / negative / gray region, so that whether a patient is infected by the mycobacterium tuberculosis or not can be more accurately diagnosed, especially aiming at a whole blood sample from a special crowd.

The utility model discloses a differential adherent cell separation device which comprises a separation box body and an upper box cover, culture cavities and a one-way conducting assembly are arranged in the separation box body, the one-way conducting assembly is arranged between the culture cavities, the side face of the separation box body extends outwards to form a supporting part, the side edge of the supporting part protrudes upwards to form a baffle ring part, and the baffle ring part is connected with the upper box cover. A limiting slot is formed among the baffle ring part, the supporting part and the separating box body, and the four sides of the upper box cover extend downwards to form side ring parts. According to the utility model, the one-way conduction between the culture cavities is realized through the one-way conduction assembly, the communication state between the culture cavities can be controlled at a proper stage according to the actual condition of cell wall attachment, the cells are ensured to flow according to the expected direction and mode, and the accuracy and purity of cell separation are effectively improved; the upper box cover and the separation box body are tightly and stably connected, the upper box cover is prevented from loosening or displacing in the using process, and the overall sealing performance and stability of the device are guaranteed.

The utility model discloses a primary cell extraction device, which relates to the technical field of cell extraction, and comprises a cell centrifugal machine and a collecting device, the bottom of the cell centrifugal machine is fixedly connected with the top of the collecting device, in the utility model, the device can automatically cut and crush a cell sample through the reverse rotation of a left group of cutters and a right group of cutters; manual intervention is greatly reduced, the cell extraction efficiency is improved, meanwhile, the crushing consistency of a packaged sample can be achieved, the purity and yield of subsequent cell extraction are improved, a cell centrifugal machine is started according to a preset separation program for primary cells, the centrifugal process is automatically completed, layering of the cells and impurities is achieved, and after centrifugation is finished, the cell extraction efficiency is improved. Separated cells and supernate can be accurately collected into corresponding containers through the collection device, tissue smashing, cell separation and collection are integrated in one device, the sample transfer frequency is reduced, the pollution risk is reduced, and the working efficiency is improved.

The invention relates to the technical field of cell culture, in particular to an efficient separation culture method for hepatocytes of small yellow croakers and a hepatocyte source identification method. The invention provides a method for efficiently separating and culturing hepatocytes of small yellow croakers, which comprises the following steps: selecting liver tissues of juvenile small yellow croakers, cleaning with a PBS (Phosphate Buffer Solution) containing high-concentration double antibodies, and digesting with pancreatin to quickly obtain hepatocytes; optimizing cell culture to obtain an L-15 culture medium containing 10% of FBS, 1% of sodium pyruvate and 0.2% of NaCl; and cell source identification is carried out by amplifying mitochondrial COI gene and 16S rRNA gene sequences. According to the method, the separation efficiency of the hepatocytes of the small yellow croakers is remarkably improved, the pollution rate is reduced, the result reliability is improved through double-gene molecular identification, the technical blank of construction of hepatocyte lines of the small yellow croakers is filled, and the method has wide application potential in the fields of aquaculture, fish physiology research, environmental toxicology detection and the like.

The present invention discloses a magnetically driven portable multi-cell separation device. According to one embodiment of the present invention, the magnetically driven portable multi-cell separation device includes: A microscope capable of nano-scale observation, A chip unit, positioned below the microscope, enabling selective cell sorting and capture, A magnetic field generator, which is adjacent to the chip unit and can control the magnetic field via a magnetic member, and A control unit, which controls the operation of the imaging unit, the chip unit, and the magnetic field generator.

The present invention relates to a method and platform for generating CAR-T cells and processing biological cells within a single functionally closed container. The method involves introducing host cells into a sealed container with multiple sterile channels, performing genetic modifications, and maintaining the cells under controlled conditions monitored by real-time feedback until a desired fraction of genetically modified cells is obtained. The container also supports target cell separation using antibody or aptamer functionalized buoyant microbubbles, facilitating the separation of target cells from non-target cells during centrifugation. Waste separation and cell harvesting are performed using pneumatic pressure, enabling high purity cell recovery. The platform includes modules for cell enrichment, sequestration, and isolation, enabling high recovery and high purity of therapeutic cell populations. This integrated closed system approach ensures sterile conditions, efficient processing, and high quality cell products for advanced therapeutic applications.

The invention relates to collagenase as well as a coding gene and application thereof. The collagenase provided by the invention is a protein (a) or (b), wherein (a) is a protein composed of an amino acid sequence as shown in SEQ ID No: 1; and (b) a protein which is obtained by substituting, deleting or adding one or more amino acids in the amino acid sequence in (a), has collagen hydrolysis or fat dissolving activity and is derived from (a). The collagenase provided by the invention can be applied to cell separation and tissue digestion, fat dissolution or collagen dissolution, and can also be applied to preparation of medicines for treating Doltt's contracture, Petroleum disease and lumbar disc herniation and degreasing in the field of beauty.

An exemplary method and system is disclosed that facilitate the integration of multiplexed single-cell impedance cytometry in a high throughput format, which can be deployed upstream from microfluidic sample preparation and / or downstream to microfluidic cell separation. In exemplary method and system may employ impedance-based quantification of cell electrophysiology on the same microfluidic chip (i.e., “on-chip”) to provide distinguishing phenotypic information on the sample, without the need for additional sample handling, preparation or dilution steps as would be needed for other flow cytometry techniques.

The application belongs to the technical field of pathological grading, and particularly relates to a lymphoma histological grading method, which comprises the following steps: screening a plurality of regions of interest rich in diagnostic information in a lymphoma pathological section image; inputting the regions of interest into a trained Hover-Net nuclear segmentation model to output a cell separation image; the cell separation image can mark the regions of different cell types, the positions of each cell and the classification information thereof; inputting the cell separation image into a classifier to output a histological grading result and the number of each cell type. The application improves the diagnostic accuracy and efficiency of follicular lymphoma pathological grading by efficiently recognizing lymphoma pathological section images of different pathological grades through a Hover-Net nuclear segmentation model.

The invention provides an autoimmune disease physical therapeutic apparatus based on a microfluidic technology and dendritic cells, and belongs to the technical field of biomedical engineering. The therapeutic apparatus provided by the invention comprises an automatic blood sampling system, a target cell separation system, a micro-fluidic chip system, a high-precision liquid flow driving system, a real-time monitoring system, a cell feedback system and an intelligent control system which are connected in sequence. The therapeutic apparatus can integrally complete the whole process from patient blood sampling, target cell separation, physical regulation and control to cell reinfusion, and solves the problem that a safe, effective and controllable autoimmune disease physical therapeutic apparatus is lacked in the prior art.

1. The name of the design product: cyclone centrifuge. 2. The use of the design product: the design product is used for reagent, solvent, chemical, cell separation, microbial sample separation and treatment. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective view.

The invention relates to the technical field of immune cells, in particular to a preparation method of human placenta-derived NK cells. The preparation method comprises the following steps: S1, separating placenta tissues; s2, placenta tissue digestion; S3, mononuclear cell separation; S4, NK cell sorting; and 5, NK cell multiplication culture.

The utility model discloses a sterile needle cylinder type spleen lymphocyte separation and extraction device, including separation part, grinding part, collection part, and separation part is needle cylinder shape, including detachable connection's upper cylinder, lower cylinder, and the lower end detachable connection cell screen of upper cylinder, and the lower end detachable connection collection part of lower cylinder, and grinding part includes push rod, rocker, and the lower end of push rod is equipped with piston head, and piston head and upper cylinder closely slide and cooperate, and the center of push rod and piston head is equipped with the through -hole of continuity, and the rocker is slid and is arranged in the through -hole, and the upper end is stretched out push rod outside and is connected the rocking handle, and the lower end is stretched out piston head outside and is connected the grinding pestle, and the lateral wall of upper cylinder is equipped with liquid adding pipe, and one end of liquid adding pipe is located in the upper cylinder, and the other end is located in the upper cylinder outside. The utility model can carry out the coherent operation of organization grinding, cell release, impurity filtration, lymphocyte enrichment under the semi -closed environment, reduces the sample pollution risk, and promotes the repeatability of experiment.

The application discloses a cell separation device, which comprises a centrifugal chamber and a rotating driving part for driving the rotation of the centrifugal chamber, a joint assembly is arranged on the centrifugal chamber, a liquid inlet interface, a waste outlet interface and a suction interface are connected to the joint assembly, a flow guide seat is arranged in the centrifugal chamber, a liquid inlet channel, a waste outlet channel and a suction channel are arranged on the flow guide seat, the liquid inlet interface is communicated with the liquid inlet channel through a first connecting pipe, the waste outlet interface is communicated with the waste outlet channel through a second connecting pipe, the suction interface is communicated with the suction channel through a third connecting pipe, and the suction channel extends to the end of the centrifugal chamber. A continuous separation method comprises the following steps: a) filling a buffer solution; b) filling a liquid containing target cells; c) cell layering; d) cell separation; or d') target cell washing. The application has the advantages of simple structure, compactness, continuous centrifugal separation, and the ability to process a larger volume of liquid.

The present application belongs to the technical field of tumor detection and cell separation, and particularly relates to a kit for separating and recovering after immunocapture of circulating tumor cells, a preparation method and application. The kit comprises an immunocapture module, a dissociation and recovery module, a cell protection module and elution and supplement factor solutions. The immunocapture module adopts magnetic microspheres modified by double antibody fusion proteins, can recognize multiple markers such as EpCAM and Vimentin, and realizes efficient capture of heterogeneous CTCs. The dissociation and recovery module uses a low-temperature buffer system and a cleavable cross-linking agent to release cells gently. The cell protection module maintains the integrity and activity of the cell membrane during the separation process. The method of the present application can ensure the capture efficiency while obtaining high-activity CTCs, and is suitable for single-cell sequencing, drug sensitivity analysis, organoid culture and patient-derived cell line construction and other preclinical research.

The present disclosure describes a sperm separation device (1). The device comprises a sample zone (5) configured to receive semen, a collection zone (15) fluidly connected to the sample zone by a migration zone (10). A cover (20) is disposed over portions of the migration region and the collection region, where the migration region comprises a groove for guiding sperm in the seminal fluid, and where sperm in the seminal fluid migrates to the collection region via the groove.

The invention relates to the technical field of cell separation and purification and cell biological detection, in particular to a method for separating and purifying fetal NRBC from peripheral blood of a pregnant woman. The invention provides a method for separating and purifying fetal NRBC from peripheral blood of a pregnant woman, which is characterized in that a LukSF and LukAB bi-component perforation toxin is designed and utilized to specifically recognize and split a CXCR1 / 2 chemotactic factor receptor and a CD11b / CD18 integrin receptor on the surface of maternal leukocyte, and the fetal NRBC is completely reserved due to lack of corresponding receptors, so that the fetal NRBC can be separated and purified. Therefore, leukocyte pollution in nucleated red blood cells of a peripheral blood fetus of a pregnant woman is efficiently removed. According to the method, the purity of the extracted fetal NRBC can be improved, the false positive of gene detection is greatly reduced, the activity and integrity of the NRBC are reserved, and the accurate diagnosis requirements of chromosome karyotype analysis, monogenic disease detection and the like are met; the method is simple to operate and low in cost, can be directly adapted to existing clinical laboratory equipment, and has large-scale popularization value.

This invention discloses a method for obtaining high-purity NK cells using apheresis, belonging to the field of cell culture. The method includes the following steps: (1) obtaining peripheral blood mononuclear cells using the apheresis blood component method; (2) adding NK cell sorting reagent to the peripheral blood mononuclear cells, mixing evenly, and allowing to stand at room temperature to obtain a cell mixture; then adding the cell mixture to the upper layer of lymphocyte separation medium, centrifuging, removing the white membrane layer and above the transparent plasma, washing the harvested cell fluid to obtain enriched NK cells; (3) purifying using CD56 positively selected magnetic beads to obtain purified NK cells; (4) inoculating the purified NK cells into a culture container coated with CD16 antibody and allowing to stand. This method can obtain NK cells with a purity of over 99%, and the cells can expand hundreds of times after 17 days of in vitro culture, exhibiting strong anti-tumor function at low target ratios.

The invention discloses a tumor holoantigen nano vaccine as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The nano vaccine comprises a composite liposome, a nucleic acid TLR agonist adjuvant, a tumor cell membrane antigen and a tumor cell soluble antigen. The membrane antigen is embedded into the lipid bilayer, and the soluble antigen and the adjuvant are wrapped in the inner cavity of the lipid to form the nanoparticles with accurate structures. The preparation method comprises the following steps: splitting a tumor cell separation membrane antigen and a soluble antigen; mixing the membrane antigen and lipid to form a lipid phase, and mixing the soluble antigen and an adjuvant to form a water phase; finally, the two phases are self-assembled into the nano vaccine through a micro-fluidic chip. The two antigens are synergistically combined at the nanoscale, the problems that a single antigen is weak in immunogenicity and incomplete in coverage are solved, the synergic anti-tumor effect far better than that of a single antigen vaccine is shown in an animal model, the preparation process is universal and controllable, and an efficient platform is provided for individualized tumor immunotherapy.

The utility model provides cell separation and extraction equipment, which comprises a support (1) and an objective table (2), the objective table (2) is used for placing a bag (100), the objective table (2) is rotatably connected to the support (1), the objective table (2) comprises an objective table main body (21) and a magnet assembly (23), the magnet assembly (23) is arranged on the objective table main body (21), the surface of the objective table main body (21) is used for placing the bag (100), and the surface of the objective table main body (21) is used for placing the bag (100). A sample in the bag (100) is located in a magnetic field generated by the magnet assemblies (23), the magnet assemblies (23) are linearly arranged according to a Halbach array, the magnetic field of the working face, facing the bag (100), of the objective table body (21) is uniform, and the magnetic field intensity of the working face (212), facing the bag (100), of the objective table body (21) is larger than the magnetic field intensity of the back face, facing away from the bag (100), of the objective table body (21).

The application discloses a method for detecting immune killing activity of whole blood without cell separation, and belongs to the technical field of in-vitro immune detection. The method directly adopts fresh peripheral blood anticoagulated by heparin, does not need density gradient centrifugation, immune magnetic bead sorting or any immune cell purification step, and directly obtains the specific killing rate of whole blood immune cells to target cells through cytokine pre-activation, co-culture of fluorescently labeled target cells, apoptosis staining and flow cytometry analysis. The application has the advantages of simple operation, short time consumption, complete cell activity reservation, high physiological correlation, good repeatability and the like, and can truly reflect the comprehensive killing function of whole blood immune cells.

This invention discloses an automated sample loading system for cell separation, comprising: a pump device, a reagent switching device, a sample container, a first solenoid valve, a second solenoid valve, a third solenoid valve, a weighing sensor, a bubble sensor, a controller, and a microfluidic chip. Sample loading is achieved through the pump device, reagent switching device, microfluidic chip, rare cell tube, sample tube, and sheath fluid tube, eliminating the need for pipettes and significantly reducing consumable usage. The sample loading process does not require frequent pipette tip changes; only the reagent switching device needs to be controlled. The response time is reduced from fractions of a second to milliseconds, improving efficiency. Furthermore, no space is required for pipetting routes or pipette tip boxes, resulting in a compact structure that facilitates device miniaturization and placement.

The invention provides a peripheral blood NK cell efficient amplification method which comprises the following steps: collecting peripheral blood and separating mononuclear cells, removing CD3 + lymphocytes by an immunomagnetic bead method to obtain initial NK cells, coating a culture plate with an anti-NKp46 antibody, performing first-stage activation under the combination of IL-2 and IL-12, and co-culturing with inactivated trophoblast cells to obtain the NK cells. Second-stage amplification is completed in a culture medium containing IL-15, IL-21, IL-33, NAC, melatonin and other additives, and finally, CD3-CD56 + cells are obtained through flow sorting; according to the method, the NK cell amplification multiple is larger than or equal to 1000 times, the purity is maintained to be 91% or above, the cell activity and killing activity are high, and an efficient and feasible scheme is provided for clinical cell therapy.

The utility model discloses a cell separation device with a function of protecting cell samples, which relates to the technical field of cell separation, and comprises a handle, the bottom of the handle is fixedly connected with a cover plate component, the outer surface of the bottom of the cover plate component is in threaded connection with a separation component, the front surface of the separation component is provided with a controller, and the controller is in threaded connection with the separation component. The separation assembly comprises a separation box, a driving motor is fixedly mounted in the middle of the bottom of a groove in the top of the separation box, the output end of the driving motor is fixedly connected with a centrifugal placement assembly, and a plurality of test tube assemblies are arranged at the bottom of the inner wall of the centrifugal placement assembly. According to the cell separation device with the cell sample protection function, the rubber layer connected with the inner cavity of the placing hole is used for extruding and clamping the test tube assembly, and the cover plate assembly is in threaded connection with the inner wall of the top of the placing box, so that the bottom of the cover plate is propped against the test tube assembly, and the test tube assembly is fixed; the test tube assembly is prevented from shaking during separation, so that the cell separation effect is not influenced.

The invention discloses a co-culture method for extracting NK from placenta blood and stimulating by using NK92 cells, which comprises the following steps: 1, separating mononuclear cells: separating the placenta blood by using a lymphocyte separating medium through a density gradient centrifugation method to obtain the mononuclear cells; 2, carrying out induced amplification on the NK cells; according to the method, placenta blood NK cells subjected to primary culture and irradiated NK92 cells are inoculated into a culture medium to start co-culture, in addition, the NK92 cells are subjected to gene editing, genes related to tumorigenicity are knocked out, the tumorigenicity eliminating effect of the NK92 cells is improved, in addition, in a placenta blood NK cell and NK92 cell co-culture system, the placenta blood NK cells and the NK92 cells are co-cultured, and the placenta blood NK cells and the NK92 cells are co-cultured. The cell proportion, culture medium components, culture temperature and humidity are controlled, a standardized co-culture process is established, the influence on the growth, proliferation and functions of the cells is avoided, and the stability of a placenta blood NK cell and NK92 cell co-culture system is ensured.

The invention discloses a duck primary hepatocyte separation and culture method based on multi-point puncture perfusion, and belongs to the technical field of cell separation and culture. The method comprises the following steps: sequentially and repeatedly puncturing and perfusing a perfusate A, a perfusate B and a perfusate C to duck liver tissues at multiple points, and then carrying out multi-step centrifugal resuspension treatment, so as to separate the duck primary hepatocytes. Experimental results show that the duck primary hepatocytes obtained by the isolated culture method have the advantages of high survival rate, complete morphological structure, high purity and the like. The simple, economical and efficient duck primary hepatocyte separation culture method is established, complex experimental devices and high-difficulty experimental operation are not needed, and the size of the taken duck liver tissue can be flexibly adjusted according to needs, so that the duck primary hepatocytes with high purity and high survival rate are obtained. The invention lays a foundation for efficient research and development of special drugs for ducks.

The application discloses a TIL cell high-efficiency amplification method and application, and belongs to the technical field of cell biology. In the initial culture stage, a specific concentration of IL-2, IL-15 and anti-CD3 monoclonal antibody combination is adopted, and autologous serum is added, so that the TIL cells can be effectively activated and initial proliferation of the TIL cells is promoted. In the large-scale amplification stage, IL-7, IL-21 and mesenchymal stem cell conditioned medium are introduced; the mesenchymal stem cell conditioned medium contains various nutritional components and growth factors, and the mesenchymal stem cell conditioned medium and the cytokines synergistically act, so that the amplification efficiency of the TIL cells is significantly improved; compared with a traditional method, the amplification multiple can be increased by 3-5 times, and the culture cycle is shortened by 5-7 days. Through optimization of a cell separation step, improvement of a culture system and regulation of a culture condition, the amplification efficiency, activity and killing capacity of the TIL cells are significantly improved, and the culture cycle is shortened.

The invention discloses a TIL cell efficient amplification method and application, and belongs to the technical field of cell biology. In the initial culture stage, a combination of IL-2, IL-15 and anti-CD3 monoclonal antibodies with specific concentrations is adopted, and autoserum is added, so that TIL cells can be effectively activated, and initial proliferation of the TIL cells is promoted. In the large-scale amplification stage, IL-7, IL-21 and a mesenchymal stem cell conditioned culture medium are introduced, the mesenchymal stem cell conditioned culture medium contains various nutritional ingredients and growth factors and has a synergistic effect with cell factors, the amplification efficiency of the TIL cells is remarkably improved, and compared with a traditional method, the amplification multiple can be improved by 3-5 times, and the culture period is shortened by 5-7 days. By optimizing the cell separation steps, improving the culture system and regulating and controlling the culture conditions, the amplification efficiency, activity and killing ability of the TIL cells are remarkably improved, and the culture period is shortened.