52 results about "Cell dissociation" patented technology

The present invention relates to new plasma or new platelet-rich plasma preparations, new cell dissociation methods, new cell associations or compositions, a method of preparation thereof, a use thereof, devices for the preparation thereof and preparations containing such a platelet-rich plasma preparation and cell associations or compositions. Specifically, the invention provides compositions comprising plasma or platelet-rich plasma alone or in combination with cell preparations for use in tissue regeneration and bone regeneration and pain reduction.

The present invention relates to new plasma or new platelet-rich plasma preparations, new cell dissociation methods, new cell associations or compositions, a method of preparation thereof, a use thereof, devices for the preparation thereof and preparations containing such a platelet-rich plasma preparation and cell associations or compositions. Specifically, the invention provides compositions comprising plasma or platelet-rich plasma alone or in combination with cell preparations for use in tissue regeneration and bone regeneration and pain reduction.

The invention discloses a biochip and a preparation method thereof. The biochip comprises a centrifugal separator, an enricher and an injection port, wherein, the centrifugal separator and the enricher are respectively provided with a discharge port; the centrifugal separator is provided with a centrifugal separator outlet, the enricher is provided with an enricher inlet, and the centrifugal separator outlet is connected with the enricher inlet; the centrifugal separator comprises a vortex microfluid channel and a plurality of micro- columns arranged in the microfluid channel along the fluid flow direction; and the micro- columns divide the microfluid channel into an inner flow channel and an outer flow channel. The biochip of the invention has the advantages of compact structure, reducedtotal area of the biochip, improved separation efficiency, short separation and enrichment process, supported in situ culture and reduced cell dissociation times; and the biochip can also be used forseparating other functional particles.

The invention provides a cell-factory-based measles virus stock solution and a measles-series attenuated live vaccine preparation. A preparation method of the measles virus stock solution includes selecting SPF chick-embryo cells, and adding cell culture fluid to prepare cell suspension liquid, and adding the cell suspension liquid into a cell factory; inoculating the cell factory with working seeds of measles viruses and the cell suspension liquid according to the ratio of the working seeds to the cell suspension liquid being (0.005-0.05):1, and standing and cultivating the cell factory at the temperature of 33+ / -1 DEG C for 3-4 days, pouring out archeocyte culture fluid, and replacing with fresh cell growth liquid for continuous cultivation; during cytopathy, collecting single measles virus liquid step by step. In an equal production scale, the batch number of cell dissociation is reduced, and the high-titer measles virus liquid is obtained, so that quality uniformity of measles-series vaccine products is improved effectively, and product yield is increased.

The invention discloses a method for culturing lung cancer primary cells, and provides a lung cancer solid tumor primary cell culture method and a matched reagent. The technology is characterized by including the steps that (1) lung cancer solid tumor tissue is treated by using a mild cell dissociation reagent, and the activity of cancer cells in the tissue is ensured to the largest extent; (2) aspecial serum-free culture medium is prepared, in-vitro culture is conducted on the tumor cells derived from a lung cancer solid tumor source through a suspension culture system, and interference of normal cells is eliminated to the largest extent while normal amplification of the cancer cells is ensured. A lung cancer primary cell culture obtained through the method can be used for in-vitro experiments of various cell levels, secondary sequencing, construction of animal models, construction of cell lines and the like. It is contemplated that the culture method has wide application prospects in the fields of lung cancer research and clinical diagnosis and treatment.

The invention relates to an isolated culture method of chicken intestinal tract epithelial GammaDeltaT cells. Physical grinding is matched with a chemical enzyme process to dissociate chicken intestinal tract epithelial cells, thereby enhancing the intestinal tract cell dissociation degree, shortening the cell treatment time and ensuring the high activity of the cells; the use of the cell separation solution with the optimum density ensures the purification of the intestinal tract epithelial lymphocytes; the magnetic bead secondary antibody separation process is utilized to ensure the higher purity of the target cells; and the cell culture detects that the purity of the target cells obtained by isolated culture is up to 90%. The method overcomes the defects in the isolated culture process of the chicken intestinal tract epithelial GammaDeltaT cells, thereby laying the foundation for chicken intestinal tract epithelial GammaDeltaT cell culture and biological and immunological researches thereof.

The invention provides a kit and method for fast separating bone marrow mesenchymal stem cells. The kit comprises a cleaning solution, a cell culture solution A, a cell culture solution B and a cell dissociation solution, wherein the cleaning solution is a phosphate buffer solution; the cell culture solution A is an alpha-MEM liquid culture medium; the cell culture solution B is fetal calf serum; and the cell dissociation solution A is a trypsin solution with the mass / volume percentage concentration of 0.25%. Compared with the traditional separation culture method, the method comprises the following steps of carrying out adherent culture for 24 hours with the culture solutions by adopting the kit and utilizing bone marrow emigrated cells and then culturing and purifying in a half solution changing way, therefore, obtained cells have high purity and are faster to proliferate and primary cell culture can be completed for 7 days generally; and in addition, primary cells are subjected to passage and cultured by utilizing the culture solutions, thus the cell density can reach 90 percent after 2-3 days, and rich sources are provided for carrying out further experimental research by utilizing the bone marrow mesenchymal stem cells.

Disclosed is a method for the dissociation of cells. Cells are processed under conditions of pH, temperature, and shear to thereby yield a mixture of cell wall ghosts and cytoplasm. Preferably, the cells are jet cooked at an alkaline pH to form an intermediate mixture, and the intermediate mixture is subsequently subjected to mechanical homogenization. Generally, the cells become dissociated, whereby at least one separate cell wall component is substantially separate from the dissociated cell walls.

The invention discloses a method for culturing gastric cancer solid tumor primary cells. The invention provides the method for culturing the gastric cancer solid tumor primary cells and an auxiliary reagent. The core of the technology is as follows: (1) treating gastric cancer solid tumor tissues by using a mild cell dissociation reagent, so as to ensure the vitality of cancer cells in the tissuesto the greatest extent; and (2) preparing a special serum-free culture medium, and carrying out in-vitro culture on tumor cells derived from gastric cancer solid tumors by utilizing a suspension culture system, so as to eliminate the interference of normal cells to the greatest extent while the normal amplification of the cancer cells is ensured. The gastric cancer primary cell culture obtained by the method disclosed by the invention can be used for in-vitro experiments of various cell levels, next-generation sequencing, animal model construction, cell line construction and the like. Predictably, the culture method has wide application prospects in the fields of gastric cancer research and clinical diagnosis and treatment.

The invention discloses a cryopreservation method for adherent culture of cells. The method comprises directly adding refrigerating fluid in a cell culture bottle, then cooling the culture bottle step by step, finally placing the culture bottle in an ultra-low-temperature refrigerator to store the culture bottle, storing the culture bottle for 2-3 months, unfreezing the cells when culture conditions permit, and continuing culture. Freezing and unfreezing steps are simple and rapid, the steps of cell dissociation and centrifugation and the like in a traditional freezing method are omitted, cell pollution and damage to cell viability are greatly reduced, the influence of adherence degree of the cells is avoided during freezing, the adherence degree of the cells can be 20-30%, limitation of traditional freezing to the number of the cells is avoided, and the cryopreservation method is extremely suitable for dealing with destructive damage which sudden conditions in a laboratory may cause to cell culture. A cell cryopreservation pipe does not need purchasing, the original culture bottle is directly utilized to carry out freezing, the cells do not need to be inoculated to a new culture bottle after unfreezing, culture fluid is added to the original culture bottle to continue culture, and the testing expenses are greatly reduced.

The invention discloses a culture method of gynecological tumor primary cells. The invention provides a gynecological tumor primary cell culture method and a matched reagent. The core of the technology is as follows: a gynecological tumor solid tumor tissue is treated with a mild cell dissociation reagent, so that the activity of tumor cells in the tissue is ensured to the maximum extent; and a special serum-free culture medium is prepared, and solid tumor cells from a gynecological tumor are cultured in vitro by using a suspension culture system, so that interference of normal cells is eliminated to the maximum extent while normal amplification of the tumor cells is guaranteed. A gynecological tumor primary cell culture obtained by the method can be used for various cell-level in-vitro experiments, second-generation sequencing, animal model construction, cell line construction and the like. It is foreseeable that the culture method has wide application prospects in the fields of gynecological tumor research and clinical diagnosis treatment.

The invention belongs to the technical field of manufacturing of lithium-ion batteries, and particularly relates to a mechanism for dissociating a battery cell winding needle and a needle dissociating method for the mechanism. The mechanism comprises a first discharge clamp, a second discharge clamp, a finger-clamping connector, a finger-clamping cylinder, a fixed seat, a photoelectric control unit and a limiting control unit, wherein the second discharge clamp is arranged corresponding to the first discharge clamp; the fixed seat is arranged between the finger-clamping connector and the finger-clamping cylinder; the photoelectric control unit and the limiting control unit are arranged on the fixed seat; the first discharge clamp and the second discharge clamp are connected with the finger-clamping cylinder through the finger-clamping connector respectively; vacuum suction holes are formed in the first discharge clamp; vacuum suction nozzles are arranged on the second discharge clamp; and the vacuum suction holes are arranged corresponding to the vacuum suction nozzles. Compared with the prior art, the yield of a battery cell can be improved; the problem that an isolating film is scrapped due to cell dissociation can be reduced; and the condition that a winding machine further works after a needle dissociating failure of the winded cell is avoided.

This invention discloses a kit used for preparing marrow desmohemoblast stem cell. This kit includes dilution, eluant, separating medium which consist of separating medium A and separating medium B, cell culture fluid A, cell culture fluid B and cell dissociation buffer A; the stated dilution is isotonic Na chloride, PBS or Hank's liquid; eluant is isotonic Na chloride, PBS or Hank's liquid; separating medium A is hydroxyethyl starch solution whose quality / volume percentage concentration is 5.5-6.5% or cellogran solution whose quality / volume percentage concentration is 0.45-0.55%; separating medium B is Percoll liquid whose density is 1.073g / mL or ficoll- hypaque solution whose density is 1.077g / mL; cell culture fluid A is low carbohydrates DMEM bouillon or DF12 nutritive medium; cell culture fluid B is fetal calf serum, new-born calf serum or calf serum; cell dissociation buffer A is parenzyme liquid whose quality / volume percentage concentration is 0.4-0.6%.

The invention discloses a culture method of colorectal cancer primary cells. The invention provides the culture method of colorectal cancer primary cells and an auxiliary reagent. The core of the method comprises the following steps: (1) treating colorectal cancer solid tumor tissues by using a mild cell dissociation reagent to ensure the vitality of cancer cells in the tissues to the greatest extent; and (2) preparing a special serum-free culture medium, and carrying out in-vitro culture on tumor cells derived from colorectal cancer solid tumors by utilizing a suspension culture system, so that the interference of normal cells is eliminated to the greatest extent while the normal amplification of cancer cells is ensured. The colorectal cancer primary cell culture obtained by the method can be used for in-vitro experiments of various cell levels, next-generation sequencing, animal model construction, cell line construction and the like. Predictably, the culture method has wide application prospects in the fields of colorectal cancer research and clinical diagnosis and treatment.

The invention discloses a lung cancer micro-tumor cell model culture method. The invention provides a novel lung cancer micro-tumor model culture technology and matched reagents, and the core of the technology is as follows: (1) using a mild cell dissociation reagent for treating lung cancer solid tumor tissues, so as to ensure the vitality of various types of cells in the tissues to the greatest extent; and (2) preparing a special serum-free culture medium, and utilizing a suspension culture system to enable various types of cells separated from lung cancer tissues to be self-assembled to form a cell mass structure with various cell components, namely a lung cancer micro-tumor model. The lung cancer micro-tumor model obtained by the method can accurately reflect various characteristics of the original focus of a patient, and is a good scientific research experiment model and a preclinical experiment model in the field of precise tumor diagnosis and treatment. It can be foreseen that the culture method has wide application prospects in the fields of lung cancer research and clinical diagnosis and treatment.

Disclosed is a method for the dissociation of cells. Cells are processed under conditions of pH, temperature, and shear to thereby yield a mixture of cell wall ghosts and cytoplasm. Preferably, the cells are jet cooked at an alkaline pH to form an intermediate mixture, and the intermediate mixture is subsequently jet cooked. Generally, the cells become dissociated, whereby at least one separate cell wall component is substantially separate from the dissociated cell walls.

The invention discloses a bovine rumen epithelial tissue dissociation method for single cell sequencing. The bovine rumen epithelial tissue dissociation method comprises the following steps of: mechanically separating rumen tissues, performing enzyme digestion treatment, filtering impurities, cleaning cells, filtering again, observing the cells, controlling the quality, removing dead cells and the like. By adopting the bovine rumen epithelial tissue dissociation method, a rumen tissue single cell suspension with high activity, high cell count and low pollution can be obtained, the yield of effective single cell count and the quality of single cell nucleic acid are ensured, and a foundation is laid for single cell sequencing and data analysis; and the problem that a bovine rumen single cell dissociation technology is not systematically perfected in the prior art is solved.

The invention discloses a culture method of a gastric cancer micro-tumor cell model. The invention provides a novel gastric cancer micro-tumor model culture technology and a matched reagent, and the core of the technology is as follows: (1) using a mild cell dissociation reagent for treating gastric cancer solid tumor tissues, so that the vitality of various types of cells in the tissues is ensured to the greatest extent; and (2) preparing a special serum-free culture medium, and self-assembling various types of cells separated from gastric cancer tissues by using a suspension culture system to form a cell mass structure with various cell components, which is called as the gastric cancer micro-tumor model. The gastric cancer micro-tumor model obtained by the method can accurately reflect various characteristics of the original focus of a patient, and is a good scientific research experiment model and a preclinical experiment model in the field of precise tumor diagnosis and treatment. It can be foreseen that the culture method has wide application prospects in the fields of gastric cancer research and clinical diagnosis and treatment.

The invention discloses a gene recombination trypsin cell dissociation solution. The gene recombination trypsin cell dissociation solution comprises the following components according to a concentration unit: 50-70mg / L of dibasic potassium phosphate, 350-450mg / L of potassium chloride, 300-400mg / L of sodium hydrogen carbonate, 7000-9000mg / L of sodium chloride, 80-100mg / L of disodium hydrogen phosphate heptahydrate, 1000-2000mg / L of glucose, 200-500mg / L of EDTA-4Na, 1-60mg / L of gene recombination trypsin and the balance of water. The cell dissociation capability of the gene recombination trypsin cell dissociation solution is superior to that of the conventional trypsin; the gene recombination trypsin cell dissociation solution is capable of completely dissociating ordinary cells within three minutes; the cell dissociation process is relatively low in damage to the cells; the cells are inoculated after the cells are completely dissociated; the cells are high in living rate and good in growth conditions; the gene recombination trypsin cell dissociation solution does not need to be preserved at the temperature of 20 DEG C below zero; the using effect of the gene recombination trypsin cell dissociation solution is not changed after the gene recombination trypsin cell dissociation solution is preserved for 12 months at the temperature of 2-8 DEG C.

The invention discloses a method for culturing porcine circovirus type 2 by a microcarrier bioreactor. The method comprises the following steps: (1) cell dissociation in a spinner bottle; (2) cell filtration; (3) cell virus receiving and tank charging culture; (4) 24 h liquid exchange; (5) gaining of a primarily received antigen; (6) second-time cell dissociation and tank charging; (7) gaining ofa secondarily received antigen and liquid exchange culture; (8) gaining of a thirdly received antigen; and (9) melting of the primarily received antigen, the secondarily received antigen and the thirdly received antigen, filtration to remove a microcarrier to obtain an antigen liquid for titer detection. By adoption of the method, the cell wall-adhering efficiency can be improved, cell aggregatesare reduced, the cell density is improved, the yield of single-batch antigens can be increased, the titer of the antigens is improved, and the production cost is lowered.

The invention discloses a separation and culture method of umbilical cord blood derived mesenchymal stem cells, and belongs to the technical field of biology. The separation and culture method comprises the following steps of performing centrifuging treatment on umbilical cord blood, taking an upper layer to obtain plasma, and taking a lower layer to obtain cells; performing inactivation treatmenton the plasma, then performing centrifuging treatment, and taking supernatant so as to obtain autoserum; mixing the lower-layer cells with normal saline to obtain a mixture, then placing the mixturein lymphocyte separation liquid, and performing centrifuging treatment so as to obtain umbilical cord blood mononuclear cells; and re-suspending the umbilical cord blood mononuclear cells with serum-free culture mediums, placing the re-suspended umbilical cord blood mononuclear cells in an autoserum coating culture dish, performing primary culture to obtain primary mesenchymal stem cells, adding cell dissociation liquid into the primary mesenchymal stem cells, performing cell dissociation treatment, and then performing subculturing. According to the method, penicllin-streptomycin for cell culture is not added in a separation process and a culture process, so that toxicity is not generated on the cells. In addition, according to the method, the autoserum coating culture dish for centrifugalseparation is adopted, so that animal derived viruses cannot be introduced.

The present invention provides a method for generation of a cell composition of mesencephalic dopaminergic progenitor cells from a starting cell composition comprising pluripotent and / or multipotent stem cells, the method comprising the steps of a) differentiating said pluripotent and / or multipotent stem cells into mesencephalic dopaminergic progenitor cells, thereby generating a cell population comprising mesencephalic dopaminergic progenitor cells and other cells, b) dissociating the differentiated cells of step a) into a single cell suspension, and c) enriching said mesencephalic dopaminergic progenitor cells by using an antigen binding molecule specific for the CD47 antigen for positive selection of said mesencephalic dopaminergic progenitor cells in said single cell suspension. Said method may be performed in a closed cell sample processing system and may be performed in an automated manner.

The invention discloses a cryopreservation method for adherent culture of cells. The method comprises directly adding refrigerating fluid in a cell culture bottle, then cooling the culture bottle step by step, finally placing the culture bottle in an ultra-low-temperature refrigerator to store the culture bottle, storing the culture bottle for 2-3 months, unfreezing the cells when culture conditions permit, and continuing culture. Freezing and unfreezing steps are simple and rapid, the steps of cell dissociation and centrifugation and the like in a traditional freezing method are omitted, cell pollution and damage to cell viability are greatly reduced, the influence of adherence degree of the cells is avoided during freezing, the adherence degree of the cells can be 20-30%, limitation of traditional freezing to the number of the cells is avoided, and the cryopreservation method is extremely suitable for dealing with destructive damage which sudden conditions in a laboratory may cause to cell culture. A cell cryopreservation pipe does not need purchasing, the original culture bottle is directly utilized to carry out freezing, the cells do not need to be inoculated to a new culture bottle after unfreezing, culture fluid is added to the original culture bottle to continue culture, and the testing expenses are greatly reduced.

The invention discloses a preparation method of an animal nervous system endothelial cell single-cell suspension, and belongs to the technical field of biology. The method comprises the following steps: carrying out mixed culture on chopped fresh nervous tissues and a cell dissociation digestion solution, centrifuging, adding an endothelial cell culture solution, and filtering to obtain a cell mixture; and mixing the cell mixture with a cell gradient separation medium, and purifying and separating the endothelial cells. The method is easy and convenient to operate, low in cost, high in efficiency and suitable for industrial production, a specific instrument is not needed, the neural endothelial cell single cell prepared through the method can be used for single cell sequencing or other analysis, and deep research on the characteristics of the neural endothelial cell is facilitated.

The methods of rare cells collection from enclosed micro filter system are provided. This invention includes surface treatment of the filter, application of cells dissociation reagent, shaking / vibration and back / side flush systems.