28 results about "Cell Pellet" patented technology

The invention belongs to the technical field of exosomes, and discloses an exosome preparation method and application, the exosome preparation method comprises the following steps: S1, culturing target cells in an induction medium, and collecting the cells and a culture solution; s2, separating the culture solution to obtain supernate A and cell precipitate; s3, performing chromatographic purification and concentration on the supernate A to obtain the exosome; in the step S1, colchicine is also added in the process of culturing the target cells to synchronize the cell cycle; a polycaprolactone-gelatin composite microcarrier is suspended in the induction culture medium, and nicotinamide, an epidermal growth factor (EGF) and pyruvic acid are contained in the induction culture medium; the method further comprises a step S30 before the step S3, resuspending the cell precipitate obtained in the step S2 with a vesicle protection solution containing trehalose, sucrose and EDTA to obtain a supernatant B, and then combining the supernatant A with the supernatant B. According to the preparation method provided by the invention, the total secretion amount of target cells is large, the intracellular vesicle-exosome precursor can be collected, and the obtained exosome is high in concentration and low in impurity content.

The invention relates to a method of producing an ester of a fatty acid and a C1-C4 alcohol, comprising successive steps of: cultivating cells of a microalga strain on a growth medium, harvesting the cultivated cells and concentrating them to obtain a wet cell pellet, suspending said wet cell pellet in an hydroalcoholic solution comprising said C1-C4 alcohol, incubating the cell suspension thus formed so as to form said ester of a fatty acid and a C1-C4 alcohol by transesterification and / or esterification of lipids produced by the microalga cells catalyzed by endogenous enzymes of said cells, and recovering said ester of a fatty acid and a C1-C4 alcohol from said cell suspension. The growth medium can advantageously be a dark fermentation effluent.

The utility model discloses a low disturbance cell collection system, including the outer tube, the inner tube is arranged in the outer tube, the inner tube is covered and has the pipe cover, wherein, the bottom of inner tube is provided with the sedimentation cavity for the cell precipitation, and the sidewall of inner tube and located above the sedimentation cavity is provided with a plurality of drainage groove, and the baffle is inserted in the drainage groove. The utility model discloses a baffle is inserted in the drainage groove sealing, and the inner tube is placed in the outer tube and pours into the centrifugal liquid and covers the pipe cover, and then is placed into the centrifugal equipment and is centrifuged. After centrifugation, the liquid is layered, and the cell is precipitated in the sedimentation cavity, and the upper layer is liquid. At this time, only need to open the pipe cover, and the baffle is pulled up, and the bottom of drainage groove is opened, and the upper layer liquid will flow into the outer tube naturally, and the cell is steady and stays in the sedimentation cavity, and solid-liquid separation is realized easily. The whole process is simple to operate, and the complicated transfer or filtering step is not needed, so that the disturbance of human operation to the cell is reduced greatly, and the integrity and activity of the cell are protected to the maximum extent.

The invention discloses a preparation method of uterine blood mesenchymal stem cells, and relates to the technical field of uterine blood mesenchymal stem cell preparation, and the preparation method comprises the following steps: filtering and diluting a uterine blood sample, adding the diluted uterine blood sample into a sample density separation medium, centrifuging, collecting an upper layer and a middle layer, repeatedly centrifuging, and taking cell precipitate to obtain primary cells of the uterine blood mesenchymal stem cells; resuspending, culturing until the fusion degree of the cells reaches 70-90%, digesting by using a recombinant trypsin solution, collecting a cell suspension, centrifuging, resuspending, inoculating and culturing until the fusion degree reaches 70-90%, digesting by using a recombinant trypsin solution, centrifuging, and resuspending; centrifuging for two times, taking cell precipitate, and resuspending by using a cell freezing medium to obtain the uterine blood mesenchymal stem cells. According to the method, after the uterine blood sample is filtered, the centrifugation step is omitted, and the uterine blood sample is directly added to the upper layer of the sample density separation liquid for centrifugal extraction, so that the normal living cell proportion of primary cells and the adherence rate after inoculation are obviously improved, the production cycle is obviously shortened, and the yield of the primary cells is improved.

A method for analyzing a tumor-specific marker and to the use of an isolate from ejaculate for diagnosing and / or monitoring prostate cancer, and / or for monitoring the success of prostate cancer therapy. The method includes the steps of performing an analysis on the isolate, wherein the tumor-specific marker selected from a protein, DNA, and RNA is analyzed, wherein the isolate was obtained by fractionating ejaculate into a low-density cell population and a high-density cell population, wherein the high-density cell population including mature sperm cells was separated and the isolate includes the low-density cell population, wherein the isolate includes a cell pellet.

The invention discloses a construction method of a tumor organoid microenvironment chip model, the model and application of the model. The construction method comprises the following steps: acquiring a tumor organ and preparing cell precipitate; carrying out enzymolysis and digestion treatment on the cell precipitate, and then adding an extracellular matrix to prepare a tumor organ suspension; the method comprises the following steps: adding a tumor organoid suspension into a micro-fluidic culture carrier, culturing to prepare suspensions of fibroblasts and vascular endothelial cells, respectively adding the two cell suspensions into corresponding hole sites of the micro-fluidic culture carrier, and finally culturing to obtain the tumor organoid microenvironment chip model. The model prepared by the method can more truly simulate the complex tumor microenvironment in vivo, is suitable for drug screening and drug sensitivity detection, is beneficial to improving the accuracy and reliability of research, and accelerates the transformation from laboratory research to clinical application.

The invention discloses a method for extracting high-purity liver cell membranes of pelteobagrus fulvidraco. The method comprises the following steps: firstly, cutting the liver into pieces to obtain large liver tissues; digesting the large liver tissue for multiple times by using isopyknic protease liquid; adding an extraction reagent 1 containing a protease inhibitor and a phosphatase inhibitor into the digested small tissues and cell precipitates, and ultrasonically crushing to obtain homogenate; performing low-speed centrifugation on the homogenate for multiple times to collect supernate, and performing high-speed centrifugation for multiple times to collect clear liquid; carrying out ultracentrifugation to collect the precipitate, and resuspending; obtaining a coarse cell membrane suspension; adding the coarse cell membrane suspension into a centrifugal tube, then adding a sucrose density gradient buffer solution until the centrifugal tube is filled with the coarse cell membrane suspension, and carrying out ultracentrifugation; and resuspending the cell membrane precipitate to obtain the pelteobagrus fulvidraco liver cell membrane. Immunoblotting verification on the cell membrane and other organelles extracted by the method shows that the cell membrane extracted by the method conforms to the expected effect of high purity, and other organelles contained in the cell membrane have few impurities.

This invention relates to the field of biomedical technology and provides a controllable cryogenic treatment mesenchymal stem cell preparation, its preparation method, and its application. The method includes the following steps: cell preparation and cryopreservation equilibration: digestion and collection; cryopreservation equilibration; removal of cryoprotectant and washing: centrifugation to remove cryopreservation solution; PBS washing; cryogenic incubation: resuspending the final cell pellet after multiple washes in pre-cooled PBS; incubating the cell suspension at 4±0.5℃ for 72 hours±10 minutes; after incubation, removing the cell suspension from the 4℃ environment, centrifuging, and resuspending in PBS. This invention yields a ready-to-use, easily stored and transported, and functionally enhanced RT-MSCs preparation. Although the cells in this preparation have lost their proliferative and metabolic activity, they still retain the complete therapeutic functions of MSCs, such as immunomodulation, anti-fibrosis, and angiogenesis promotion.

The invention provides a lung qi and blood barrier protection effect evaluation method. Specifically, the invention provides a method for evaluating the lung qi and blood barrier protection effect of a to-be-detected medicine, which comprises the following steps: (i) providing a pulmonary alveolar lavage fluid (BALF) sample of a subject receiving the to-be-detected medicine and / or reference substance administration; (ii) extracting a cell precipitate from the BALF and lysing the cell precipitate to obtain a protein extract; (iii) dyeing the protein extract and the protein standard substance with a Prussian blue dyeing solution; (iv) measuring the absorbance at the wavelength of 680 + / -10nm by using a microplate reader; wherein a standard curve is drawn according to the absorbance of the protein standard substances with different concentrations, and the absorbance of the protein extract is substituted into the standard curve to calculate the concentration of the ferrihemicin, so that the protective effect of the medicine on the lung qi and blood barrier is judged. The method disclosed by the invention can be used for simply, quickly and accurately evaluating the lung qi and blood barrier protection effect of the to-be-detected medicine.

The utility model relates to the technical field of cell extraction, and provides a centrifugal tube suitable for cell precipitation extraction, which comprises a first centrifugal tube, a second centrifugal tube and a fixing mechanism, the first centrifugal tube is detachably connected with the second centrifugal tube through the fixing mechanism, and scale marks are carved on the outer side walls of the first centrifugal tube and the second centrifugal tube; the fixing mechanism comprises a first connecting block and a second connecting block, the first connecting block is mounted on the first centrifugal tube, the second connecting block is mounted on the second centrifugal tube, and a plurality of limiting pieces are mounted at the top of the second connecting block. The centrifugal tube has the beneficial effects that the centrifugal tube is divided into two parts, namely the upper tube body and the lower tube body, and the two parts are detachably connected by adopting the fixing mechanism, so that the centrifugal tube can be easily assembled and disassembled, and the upper tube body can be slightly rotated to be separated from the lower tube body in cell precipitation extraction work, so that the extraction depth is reduced, and the extraction efficiency is improved. The bottom sediment can be more easily and completely taken out, and the cell sediment extraction work is facilitated.

The application belongs to the technical field of pharmaceutical analysis, and provides a method for quantitatively determining the concentration of PROTAC drugs in cells with high specificity, comprising the following steps: (1) selecting a cell strain expressing a PROTAC drug target; (2) culturing the cell strain to the logarithmic growth phase; (3) inoculating the cell suspension on a cell culture plate for culture; (4) adding the PROTAC drug to the cell culture plate, mixing and continuing to incubate; (5) removing the cell culture medium in the cell culture plate; dispersing the cell culture medium into a single cell suspension, centrifuging to discard the supernatant and obtain a cell precipitate; (6) lysing the cell precipitate, collecting the supernatant by ultracentrifugation, and performing ultra-high performance liquid chromatography-tandem mass spectrometry analysis; (7) preparing a blank cell suspension and a series of standard solutions of PROTAC drug concentrations, performing ultra-high performance liquid chromatography-tandem mass spectrometry analysis, and constructing a standard curve; and (8) using the standard curve to obtain the concentration of the PROTAC drug in the cells.

The present application relates to the technical field of medicine, and more particularly to a boronized cell membrane and a preparation method and application thereof.The preparation method of the boronized cell membrane comprises the following steps: adding tumor cells into a culture medium containing a monosaccharide-modified boron drug to perform incubation, collecting cell precipitates after the incubation is completed, and then extracting cell membranes, so that the boronized cell membrane is obtained.The present application provides a boronized cell membrane and a preparation method and application thereof.By using the monosaccharide-modified BSH, the obtained boron drug is anchored on the tumor cell membrane, and then the boron drug is delivered to the tumor tissue by the homologous targeting property of the cell membrane itself, so that the boronized cell membrane has high biocompatibility and helps to better achieve BNCT treatment.

The invention discloses an efficient universal mouse pancreatic tissue single-cell suspension preparation method, and belongs to the technical field of cell biology. Pancreas are rich in digestive enzymes, so that the treatment difficulty of in-vitro cells is high, and a high-quality single-cell suspension is difficult to prepare by a traditional method. The method comprises the steps of reagent preparation, tissue sampling, tissue dissociation, red cracking and cleaning, AOPI dyeing quality inspection and the like. Components and concentrations of a tissue enzymatic hydrolysate, a cell cleaning solution and a cell suspension are determined in reagent preparation; the acquisition method of the pancreatic tissue of the mouse is defined by tissue sampling; the tissue dissociation adopts a two-round digestion mode, and the cut tissue is subjected to enzymolysis by using a compound enzyme solution under specific conditions; the cell precipitates are treated through red cracking and cleaning; aOPI dyeing quality inspection is used for analyzing cell count and motility rate. According to the method, the culture medium containing specific components and the compound enzyme liquid are creatively used, the problem that pancreas single cells are difficult to dissociate can be effectively solved, the single-cell suspension with high motility rate, high purity and low adhesion is obtained, and the requirements of downstream single-cell experiments are met.

The application discloses a quality control product for human papillomavirus molecular detection and a preparation method thereof, and belongs to the technical field of biological products. After synthesizing 25 types of HPV full genomes of high-risk types (HPV16, HPV18, HPV26, HPV31, HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV53, HPV56, HPV58, HPV59, HPV66, HPV68, HPV73, HPV82) and low-risk types (HPV6, HPV11, HPV42, HPV43, HPV44, HPV81, HPV83), the HPV full genomes are connected into a pcDNA3.1(+) skeleton vector system, the HPV full genomes are integrated into the genome of a human ovarian cancer cell line ovcar3 respectively, and 25 strains of human ovarian cancer cell lines stably integrating the HPV full genomes are obtained. The new quality control product or kit of various types of simulated clinical samples such as genomic DNA, cell precipitate and cell suspension can be prepared and is suitable for various detection platforms. The human cell line quality control product stably integrating the full-length genomes of the 25 HPV types can more accurately determine the viral load and control the whole process of HPV detection.

The utility model discloses a centrifuge tube for cell precipitation, which comprises a centrifuge tube body, the top of the centrifuge tube body is connected with a tube cover through threads, an inner strip is arranged in the centrifuge tube body, one side surface of the inner strip is attached to the inner wall of the centrifuge tube body, the other side of the inner strip is connected with an inner shaft, and the inner shaft is connected with a tube cover. A rotating shaft is rotationally embedded in the pipe cover, a rotating block is fixed to the top of the rotating shaft, and the top end of the inner shaft is fixed to the surface of the bottom of the rotating shaft; the rotating block, the rotating shaft, the inner shaft and the inner strips are designed, the rotating block is held to rotate the rotating shaft, the rotating shaft drives the inner strips on the inner shaft to rotate together, the inner strips scrape off cell sap attached to the inner wall of the centrifugal tube body, and the situation that the cell sap is attached to the inner wall of the tube body and cannot rotate centrifugally at a high speed in the liquid, and sedimentation of the cell sap is affected is avoided; and the inner shaft is straightened by the supporting sleeve, so that the inner shaft is prevented from skewing.

This invention belongs to the field of biotechnology. Specifically, this invention discloses neural stem cells, their preparation methods and applications, including the following steps: (1) washing hippocampal tissue with PBS buffer, placing the washed hippocampal tissue in a digestive solution for digestion, adding a digestion termination solution to terminate digestion, centrifuging to obtain cell pellet, washing the cell pellet with PBS buffer, centrifuging, resuspending the cells in DMEM / F12 medium, and adjusting the cell concentration to 1×10⁻⁶. 4 (2) Primary culture; (3) Amplification culture. The amplification culture medium described in this invention can provide a carrier with viscosity during the culture of neural stem cells, promote the division and proliferation of neural stem cells, improve the activity and stability of neural stem cells, and increase the survival rate and cell number, and has broad application prospects.

This invention provides a kit, method, and single-cell library construction method for the dissociation of marine animal tissues. The dissociation method includes: S1) dissociating marine animal tissue samples at 8℃~12℃ using a cell dissociation buffer to obtain a single-cell suspension; S2) centrifuging the single-cell suspension and washing the single-cell precipitate obtained after centrifugation with a cell resuspension buffer to obtain single cells from the marine animal tissue sample; wherein, the cell dissociation buffer comprises a protease; the protease is Bacillus subtilis protease A; the concentration of sodium ions in the cell resuspension buffer is 300-800mM, and the concentration of potassium ions is 5-20mM.

The invention relates to a method of producing an ester of a fatty acid and a C1-C4 alcohol, comprising successive steps of: cultivating cells of a microalga strain on a growth medium, harvesting the cultivated cells and concentrating them to obtain a wet cell pellet, suspending said wet cell pellet in an hydroalcoholic solution comprising said C1-C4 alcohol, incubating the cell 5 suspension thus formed so as to form said ester of a fatty acid and a C1-C4 alcohol by transesterification and / or esterification of lipids produced by the microalga cells catalyzed by endogenous enzymes of said cells, and recovering said ester of a fatty acid and a C1-C4 alcohol from said cell suspension. The growth medium can advantageously be a dark fermentation effluent.

This invention proposes a method for preparing a single-cell suspension of porcine pancreatic cells and its application. The method includes the following steps: S1, taking porcine pancreatic tissue, removing surface fat, and mincing the tissue; S2, washing the tissue fragments, and then placing the tissue into a mixed enzyme dissociation solution for the first digestion; S3, after the first digestion, collecting the supernatant, and adding the remaining tissue to the mixed enzyme dissociation solution for a second digestion; S4, after the second digestion, collecting the supernatant and combining it with the supernatant from step S3, and filtering; S5, centrifuging the filtered dissociation solution, discarding the supernatant, and resuspending the cell pellet to obtain a cell suspension; the mixed enzyme dissociation solution includes trypsin, collagenase I, elastase, lipase, DNase I, heparin, sodium sulfate, and DH Kans buffer. The enzyme mixture of this invention has the effects of removing intercellular matrix, reducing cell aggregation, protecting cell membrane morphology, and reducing cell damage.

This invention provides a method for producing full-length antibodies in Escherichia coli at a yield of approximately 200 mg / L or more. [Solution] A method for producing a full-length antibody comprising a heavy chain (HC) and a light chain (LC), comprising culturing Escherichia coli cells expressing the coding sequence of the HC and the coding sequence of the LC in a culture medium under conditions acceptable for producing the HC and the LC, wherein the full-length antibody is produced in an amount of at least about 200 mg per liter of the culture medium, or the weight percentage of the produced full-length antibody relative to the weight of the cell pellet produced from the Escherichia coli cells is in the range of 0.05% to 20%.

This invention provides a method for producing cell tissue structures that are highly stable and reproducible. [Solution] The number of cells in the cell suspension and the cell diameter are counted (S10). The cell volume is calculated from the number of cells and cell diameter obtained in the counting step (S20). A cell pellet is obtained by centrifuging the cell suspension containing the cells whose cell volume has been calculated (S30). The cell pellet volume, which is the total volume of the cell pellet, is calculated (S40). Based on the cell pellet volume, a solvent containing biomaterials is added to the cell pellet so that the cells have a desired cell volume fraction, and a mixture is obtained (S50).

The invention relates to the field of biomedicine, and discloses a construction method and application of CIC rearrangement sarcoma primary cells. The construction method comprises the following steps: acquiring a malignant pleural effusion sample of a CIC rearrangement sarcoma patient; centrifuging the pleural effusion sample, and separating to obtain cell precipitate; cleaning the cell precipitate, lysing red blood cells, and centrifuging to obtain CIC rearranged sarcoma cells; and resuspending the CIC rearranged sarcoma cells in a cell culture solution, filtering, and culturing to obtain the CIC rearranged sarcoma primary cells. The technical bottleneck of CIC rearrangement sarcoma primary modeling is successfully broken through, and the established cell line and the related culture technology thereof have important scientific research and clinical transformation values. Besides, development of an animal model constructed based on the cell line and related diagnostic reagents is expected to provide key technical support for precise diagnosis and treatment of the rare sarcoma, and the cell line has remarkable social benefits and application prospects.

The application discloses a method for preparing a cell block by using mucin emulsifier and application thereof, and the preparation method comprises the following steps: centrifuging a non-gynecological sample to obtain a first separation liquid, adding the first separation liquid into a formaldehyde-alcohol solution to centrifuge to obtain a second separation liquid, and adding the second separation liquid into mucin emulsifier to centrifuge to obtain a cell block. The preparation method is simple, the manufacturing cost is low, the cell block has high enrichment efficiency, cell loss is small, and the mucin emulsifier does not affect cell activity. The cell block prepared by using the mucin emulsifier has the following advantages: the appearance of the section of the cell block is complete, the section has a regular circular structure, the porosity is moderate, and the thickness is uniform; the application effect of the section in two kinds of staining is good: the hematoxylin-eosin staining has a clear background and clear contrast, cells are uniformly distributed in a circular area, cell morphology presents a stereoscopic effect, and the excellent rate of the stained section is 98.94%; the immunohistochemical staining has a clean background, and structures and cells are specifically dyed, and the excellent rate of the stained section is 99.30%, so that the cell block has a wide application prospect.

To provide a method for producing cell pellets that can improve the visibility of the cell pellet, which is a mass of RI-labeled cells to be obtained. [Solution] A method for producing a cell pellet according to one aspect of the present invention includes: a preparation step of preparing a cell suspension containing cells in a physiological buffer; a labeling step of labeling the cells contained in a mixture of the cell suspension and a solution of a radionuclide with the radionuclide; and a separation step of separating the mixture into a cell pellet, which is a mass of the labeled cells, and a residual RI solution containing the radionuclide that was not taken up by the cells. The diameter of the cells contained in the mixture is 10 μm or more, and the number of cells contained in the mixture is 1.2 × 10⁻⁶. 7 It is more than just cells.

The invention relates to a device for improving gene mutation cloning inoculation efficiency of mouse lymphoma cells. The device comprises a shaking table, a liquid storage box and a multi-channel inoculation assembly, a liquid storage box is mounted on the shaking table and is used for driving the liquid storage box to shake so as to keep cell suspension in the liquid storage box uniform; the liquid storage box is used for storing a cell suspension and is provided with a plurality of liquid taking holes; the multi-channel inoculation assembly comprises a plurality of pipettor suction heads; the pipettor suction head can move and be inserted into the liquid taking hole so as to suck the cell suspension in the liquid storage box. According to the invention, the liquid storage box is shaken and uniformly mixed through the shaking table, so that the continuous uniform mixing of the cell suspension is realized, the consistency of the initial cell concentration of each parallel sample is fundamentally ensured, and the operation efficiency and the consistency are improved: the cell suspension in the liquid storage box is synchronously sucked by a plurality of pipettor suction heads of the multi-channel inoculation assembly and is inoculated into a porous plate; the original manual operation in several minutes is shortened to be completed in several seconds, so that the cell precipitation difference caused by the sequence of inoculation time is avoided.

This invention relates to the field of automated sampling technology and discloses an automated sampling device for the analysis of trace components in cell culture supernatant. The device includes a sampling worktable and a first robotic arm and a second robotic arm mounted thereon. The first robotic arm has a piston pump assembly at its end, and a sampling component is detachably mounted on its bottom via a threaded interface. The sampling component includes a sample sampling cylinder and a sampling head at the bottom. A suction port is provided on the side of the sampling head, and an insertion plug is slidably connected inside. An isolation balloon with a top opening is fixedly connected to the bottom end of the insertion plug. This invention uses the sampling cylinder directly as a storage container, eliminating residual loss on the inner wall during sample transfer. The isolation balloon effectively buffers the suction force, avoiding disturbance to the cell sedimentation layer, and adapts to differences in sedimentation layer thickness in each well, ensuring the purity and consistency of simultaneous multi-well sampling. This significantly improves the accuracy, reliability, and efficiency of automated micro-sample sampling.