2041 results about "Cultured Cell Line" patented technology

The invention provides a preparation method of a cell culture three-dimensional scaffold and cell culture meat, which comprises the following steps: firstly, preparing a gelatin-carrageenan mixed solution and a gel precursor solution of transglutaminase, dividing into two parts, incubating at constant temperature to obtain two parts of composite hydrogel, randomly freeze-drying one part to obtain aerogel, and freeze-drying the aerogel to obtain the cell culture three-dimensional scaffold. Stacking on the other part and incubating at constant temperature again to obtain the prefabricated double-layer gel. The prefabricated double-layer gel is directionally frozen, ice crystals grow upwards from the bottom, freezing is stopped after the temperature is reduced to the set temperature, and the cell culture three-dimensional scaffold is obtained through drying. The stent adopts an ordered-disordered partition design, parallel micro-channels are formed in the lower layer by using a directional ice template method, and muscle fiber arrangement is simulated; a porous spongy structure is constructed on the upper layer by using a random ice template method, and is adapted to adipocyte proliferation. Two layers of interfaces are interpenetrated and communicated, natural mechanical gradients are combined, natural tissue characteristics are re-engraved, the traditional limitation is broken through, the cell co-culture efficiency and the cultured meat quality are improved, and industrialization is promoted.

The invention discloses a cord blood-derived regulatory T cell amplification culture medium and a use method thereof, and relates to the field of animal cell culture and immune cell therapy, the culture medium is a serum-free culture medium, the culture medium is composed of a basic culture medium, a serum substitute combination, a combination of four cell factors, an immunosuppressor, an apparent stabilizer and an antioxidant, and all the components play a role synergistically; according to the use method, cell amplification is realized through a collaborative process of staged environmental regulation and targeted activation. According to the method, the problems of exogenous pollution and batch difference caused by dependence of a serum-containing system on cord blood-derived regulatory T cell amplification in the prior art are solved, the defect that a serum-free scheme is difficult to consider the amplification efficiency and the function stability at the same time is overcome, the cell amplification quality and the clinical application safety are guaranteed, and the method is suitable for large-scale clinical transformation requirements.

The invention discloses a culture medium and application thereof, and a method for in-vitro separation culture and adipogenic differentiation of precursor adipocytes in goat muscle, and relates to the technical field of cell culture, the culture medium comprises the following components: 8-12 [mu] g / mL of insulin, 8-12% by volume of fetal calf serum, 95-105 U / mL of penicillin, 95-105 [mu] g / mL of streptomycin and the like. On the basis of the prior art, a proper amount of liquiritin and pilocarpine are mainly added into a culture medium, and synergistic cooperation of the liquiritin and pilocarpine is found, so that the adipogenic differentiation capacity of cells such as mesenchymal stem cells and precursor adipocytes is remarkably improved, and the application range is wide. On the basis, the invention also provides a complete set of perfect in-vitro isolated culture and adipogenic differentiation method of the goat intramuscular precursor adipocytes, and a method and a way are provided for a series of researches developed through the goat primary intramuscular precursor adipocytes in the future.

The invention relates to the technical field of biology, and particularly discloses a device and a method for separating an adherent cell through a vortex sound field, an ultrasonic module, piezoelectric ceramics and an Archimedes spiral structure concave lens are arranged to generate the vortex sound field to act on a cell culture dish, so that an adjustable shear flow is generated on the cell culture dish, and the adherent cell is separated from the cell culture dish. And locally separating the adherent cells in the cell culture dish. According to the concave lens with the Archimedes spiral structure, the problem of defocusing of a high-order vortex sound field can be solved, and the falling control accuracy of adherent cells is improved. The proposed method does not require any physical contact with adherent cells, and does not require insertion of an instrument into a cell culture dish, thereby reducing the risk of contamination and cell damage. The stable flow generated by absorbing sound energy by the liquid is used for indirectly separating the adherent cells, the method is extremely low in energy density and has good biocompatibility, and the activity of the cells is ensured while the cells are efficiently separated.

The invention provides a method for producing cell culture meat through 3D scaffold-free co-culture of pseudosciaena crocea stem cell spheres, and belongs to the technical field of preparation of cell culture meat. A method for producing cell culture meat by 3D scaffold-free co-culture of pseudosciaena crocea stem cell spheres comprises the following steps: mixing and inoculating pseudosciaena crocea myoblasts and pseudosciaena crocea precursor adipocytes into a cell culture plate subjected to anti-adhesion treatment for proliferation culture, spontaneously gathering to form cell spheres, and carrying out differentiation culture to obtain the cell culture meat. According to the co-culture mode provided by the invention, high-efficiency myogenic differentiation and adipogenic differentiation of myoblasts and precursor adipocytes of the large yellow croakers can be realized through proliferation culture and differentiation culture, and an in-vivo microenvironment can be effectively simulated, so that cultured meat similar to a traditional meat product in texture and flavor is produced.

The embodiment of the invention provides an organ chip, and relates to the technical field of organ chips, the organ chip comprises at least two function parts and a dielectric layer located between the two adjacent function parts, the at least two function parts are stacked, and each function part is provided with a cell culture chamber; the dielectric layer is used for isolating contact between cells of adjacent cell culture chambers and allowing interaction and substance exchange between signal molecules between the adjacent cell culture chambers, and through the stacked functional part and dielectric layer structure, interaction of the signal molecules is allowed while different cell culture chambers are physically isolated; the air guide part and the connector of the culture box are tightly pressed, the air pressure regulation and control precision and the culture environment stability are guaranteed, the air pressure stability in the culture box is effectively guaranteed, and external microorganisms, dust and other pollutants are effectively prevented from invading the culture environment.

A method and apparatus for calibrating an imaging system provides a program stored in the imaging system having a graphical user interface and providing a plurality of tools to align hardware components of the imaging system including cameras for imaging cell culture vessels received by the imaging system and having a tool for providing a video image output of a cell culture vessel received in the imaging system.

The invention relates to the cross technical field of biological medicine and image processing, and discloses a mesenchymal stem cell aging detection method based on image processing. The method comprises the following steps: automatically collecting bright field and multi-channel fluorescence images in an integrated cell culture monitoring device; after background correction and illumination homogenization, segmenting the cells by using a U-Net network and extracting single cell contours; calculating characteristics such as cell area, roundness, cytoplasmic ratio, nuclear form irregularity index, lysosome fluorescence intensity mean value and distribution entropy; and inputting a gradient boosting decision tree model to judge the unicellular aging state, and evaluating population aging with a 20% positive rate threshold. According to the invention, label-free, non-invasive and high-flux accurate detection is realized, and the method is superior to manual interpretation.

The invention relates to a culture medium for high-density serum-free suspension culture of BHK-21 cells and application of the culture medium. The culture medium contains inorganic salt, amino acid, vitamins, energy substances, a buffer substance, an antioxidant, sterol, polyamine, an additive and an indicator, and the antioxidant comprises dihydrolipoic acid and S-acetyl-L-glutathione. According to the invention, the culture medium for serum-free suspension culture of the BHK-21 cells, which is clear in components and remarkable in culture effect, can be provided. The added dihydrolipoic acid and S-acetyl-L-glutathione are combined as antioxidants of the culture medium, so that the survival rate of BHK-21 cells cultured in the culture medium is improved while the oxidation resistance of the serum-free suspension culture medium is improved.

The invention relates to the technical field of biology, in particular to application of active polypeptide in maintaining dryness of umbilical cord mesenchymal stem cells. The active polypeptide disclosed by the invention is PNC-27, and the amino acid sequence of the PNC-27 is as shown in SEQ ID NO: 1. Experiments find that the PCN-27 polypeptide can effectively promote proliferation of the umbilical cord mesenchymal stem cells by regulating expression of periodic proteins Cyclin-D1 and CDK1, and can also effectively maintain dryness of the umbilical cord mesenchymal stem cells by promoting expression of dry gene Oct-4 protein in the umbilical cord mesenchymal stem cells. Experiments fully prove that the PCN-27 polypeptide can effectively maintain the dryness of the umbilical cord mesenchymal stem cells and stimulate the potential application value of the umbilical cord mesenchymal stem cells in the fields of cell culture, biomedicine and the like.

The invention relates to the technical field of medicines, in particular to application of salinomycin in preparation of oral medicines for preventing or treating monkey pox virus (MPXV) infection. Salinomycin is a polyether antibiotic produced by actinomycetes. It is found for the first time that salinomycin can efficiently inhibit monkey pox virus infection on the cell culture level, and can significantly improve the survival rate of mice infected with monkey pox virus in small animal living bodies. More importantly, salinomycin can be administered by oral administration, has good bioavailability and safety, and provides a brand new treatment means for monkey pox virus infection. The medicinal value of salinomycin in resistance to monkey pox virus infection is reported for the first time at home and abroad, and the application prospect of salinomycin in the field of viral disease treatment is expanded.

A method includes introducing a suspension including cells suspended in a cell culture medium through a feed port or a drain port into a cavity of a cell culture vessel, the suspension being in an amount sufficient to cover a gas permeable, liquid impermeable membrane positioned at a bottom of the cell culture vessel, the feed port being disposed through a surface of the cell culture vessel and configured to permit additional cell culture medium into the cavity, and the drain port being disposed through the surface of the cell culture vessel and configured to permit removal of the cells, cell culture medium, and used cell culture medium from the cavity, allowing the cells to settle on the gas permeable, liquid impermeable membrane by gravity, removing the used cell culture medium through the drain port and introducing the additional cell culture medium through the feed port such that a constant volume is maintained in the cell culture vessel until the cells expand to a desired cell density, wherein the removing and introducing are performed subsequent to allowing the cells to settle on the gas permeable, liquid impermeable membrane, resuspending the cells in the cell culture medium in the cell culture vessel, wherein the resuspending is performed after the desired cell density is attained, and removing the resuspended cells and the cell culture medium through the drain port.

The invention belongs to the technical field of cell biology, and particularly discloses a culture medium and a culture method of fish primary liver cells. The culture medium of the fish primary liver cells comprises the following components: 90% of an M199 culture medium, 10% of fetal calf serum, 100 U / mL of penicillin, 0.1 mg / mL of streptomycin, 5 ng / mL of EGF, 10 [mu] U / ml of insulin and 10 mM of dexamethasone. Based on the culture medium, an efficient and stable fish liver cell culture method is developed by combining a standardized anesthesia and disinfection process, a precise dissection and liver sampling method, an optimized tissue digestion and cell separation scheme, an efficient cell purification and adherent culture system, a stable passage technology and the like; a standardized hepatocyte line establishment operation process is defined, and the problems that species binding exists in an existing fish liver cell culture method, and a standardized culture process suitable for multiple fish fingerlings is difficult to form are solved.

The utility model discloses a table type glass bioreactor and relates to the technical field of bioreactors. The bioreactor comprises a bioreactor body, wherein a sealing cover is arranged at the top of the bioreactor body. The moving mechanism can drive the stirring mechanism to rotate while moving up and down, so that the stirring mechanism is in a working state of rotating while moving up and down in the bioreactor body, and high-temperature liquid positioned below the bioreactor body can flow to an upper area; meanwhile, the liquid with lower temperature above the bioreactor body flows to the area with higher temperature below the bioreactor body, so that the temperature gradient of different height positions in the bioreactor body is broken, even in the area far away from the heating plate, the uniform distribution of the temperature can be realized, and the consistency of the temperature in the bioreactor is improved; and biological processes such as cell culture and fermentation are facilitated.

The present disclosure provides materials and methods related to the purification of viral vectors. In particular, the present disclosure provides peptides, compositions, adsorbents, and related methods, capable of removing process-related impurities (e.g., host cell proteins, nucleic acids, and media components) and product-related impurities (e.g., product fragments, product aggregates, and inactive forms derived from product degradation by or association with other species in the cell culture harvest) from samples during the production and purification of adenovirus.

The invention discloses a cat kidney suspension culture cell line suitable for serum-free suspension culture and application of the cat kidney suspension culture cell line. The name of the cell line is F81-B4, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.46350. After 50 generations of continuous subculture, the cells still can maintain the original proliferation level and cellular morphology, the cell number can reach 9.0 * 10 < 6 > / mL after the cells are subcultured at the density of 1.0 * 10 < 6 > / mL for three days, the multiplication time is 22-26 hours, and the cell proliferation speed is equivalent to that in a shake flask when the cells are amplified to a 10L bioreactor for culture. The cell line is highly susceptible to feline parvovirus, feline calicivirus, feline herpes virus, canine parvovirus and canine distemper virus, the virus content of a virus solution harvested 72 h after FPV virus inoculation can reach 107.5 TCID50 / mL, the virus content of a virus solution harvested 24 h after FCV virus inoculation can reach 1010.8 TCID50 / mL, the virus content of a virus solution harvested 48 h after FHV-1 virus inoculation can reach 107.5 TCID50 / mL, compared with an existing F81 adherent cell production process, the production process has the advantages that the production efficiency is high, and the production cost is low. The method has the characteristics of rapid lesion, short virus collection time and high titer. The cell matrix is an ideal cell matrix which can be used for culturing the three viruses.

The instant invention relates to compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, e.g., an anti-IL-1R1 antibody or antigen binding portion thereof, and methods, e.g., cell culture and protein purification methods, for producing such compositions.

Provided herein are peptide amphiphiles (PAs) comprising a bioactive peptide, nanofibers displaying the bioactive PAs, and methods of use thereof. The disclosed peptide amphiphiles comprise a hydrophobic tail, a structural peptide segment, a charged peptide segment, and a bioactive IKVAV peptide. The disclosed PAs may be used in cell culture methods and in methods of treating central nervous system injury.

The invention discloses a physiological microenvironment-based cell culture simulation system, which relates to the field of cell culture, and comprises a main control unit adopting an embedded microcontroller; the multi-mode environment simulation unit is connected with the main control unit and comprises a fluid shear force module and a mechanical stretching module; the multi-parameter environment monitoring unit is connected with the main control unit and comprises an optical sensor for monitoring cell density, a pH sensor for monitoring environment parameters and a temperature sensor; the culture chamber is used for accommodating a cell culture; the environment regulation and control unit is used for regulating culture environment parameters; the multi-modal environment simulation unit and the multi-parameter environment monitoring unit are integrated, and the main control unit optimizes a multi-target control strategy in a manner of dynamically adjusting a weight coefficient based on cell growth state parameters monitored in real time, so that accurate and adaptive closed-loop management of cell culture is realized.

The invention relates to the technical field of aquatic animal cell culture, in particular to a rapid enrichment and separation method for gonad somatic cells of plectropomus leopardus. Comprising the following steps: selecting healthy plectropomus leopardus for anesthesia; the method comprises the following steps: taking gonad tissues, performing sterile treatment, cutting, centrifugal washing and filtering, inoculating the gonad tissues into a culture medium for primary culture to obtain gonad cells emigrated from tissue blocks, performing trypsin-EDTA digestion and centrifugal washing, and treating the gonad cells with 1 * PBS containing polysorbate; after closing, antibody incubation and washing, the marking effect is verified through a fluorescence microscope, and specific enrichment of gonad somatic cells is achieved; and filtering and purifying the enriched fluorescence-labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescence-positive cells under a fluorescence visual field, transferring the fluorescence-positive cells into a culture dish containing a complete medium, inoculating the separated fluorescence-positive cells into the optimized complete medium, and carrying out purification culture, so that the cells grow in a wall-attached manner and stable passage is realized.

The invention relates to the field of synthetic biology, metabolic engineering and fermentation engineering, and discloses a genetically engineered bacterium and application thereof in production of alpha-ketoglutaric acid.The genetically engineered bacterium comprises the steps that the genetically engineered strain with escherichia coli BW25113 or corynebacterium glutamicum ATCC13032 as a host is constructed; overexpression type isocitrate dehydrogenase genes are integrated through chromosomes, alpha-ketoglutarate dehydrogenase complex genes are knocked out, NADPH invertase is overexpressed through plasmids, and exogenous AKG transporter protein is introduced, so that metabolic flux directional regulation and control and product efflux enhancement are realized; meanwhile, a staged fermentation process is adopted and comprises dynamic gradient adjustment of inorganic phosphorus concentration and two-stage pH control. According to the method, the AKG yield of 78.2 g / L can be obtained under the 5L tank scale, the sugar acid conversion rate reaches 0.68, and the method has the advantages of being high in yield, low in byproduct and controllable in cost. The obtained AKG is suitable for a plurality of fields such as medical intermediates, functional foods and cell culture additives, and has wide industrial application prospects.

The invention relates to the technical field of cell culture, and discloses a stem cell differentiation degree identification method based on image features. The method comprises the following steps: continuously capturing multi-temporal image data in a stem cell culture process through a high-resolution microscopic imaging system, and generating an image and environment synchronous data set; processing the data set, extracting dynamic morphological features and texture change modes of the stem cells, and constructing a fusion feature set; outputting a differentiation process index based on the fusion feature set; a differentiation degree grade division rule is set, and a differentiation process index and differentiation grade mapping table is established; in a real-time application stage, acquiring real-time imaging data and environment readings, and inputting the real-time imaging data and the environment readings into an evaluation network to calculate real-time differentiation process indexes; querying the mapping table according to the real-time index to obtain a differentiation grade judgment result; comparing the judgment result with a preset target range, and if the judgment result falls into the target range, automatically executing differentiation state marking and culture parameter adjusting operation.

The invention relates to the technical field of biology, in particular to an MDCK cell strain and application of the MDCK cell strain in preparation of influenza vaccines. The invention provides an MDCK cell strain, the preservation number of which is CCTCC NO: C202504. The cell strain not only can maintain high sensitivity and high yield to influenza viruses and ensure the yield and quality of vaccines, but also can remarkably improve the biological safety of cell matrix influenza vaccines due to elimination of tumorigenicity risk, and has a good application prospect. And a key technical support is provided for promoting domestic research and development and marketing of influenza vaccines based on cell culture.

Provided are methods for the creation of ultra-high density cryopreserved cell banks. In certain embodiments, these methods employ altered perfusion culture techniques that allow for production of ultra-high density cell cultures that can be cryopreserved at unexpectedly high cell densities without the need for any cell concentration steps, while retaining excellent cell viability and quality.

A sequential cell culture system and methods for the in vitro generation of human spermatozoa from differentiating spermatogonia (dSPGs) and spermatogonial stem cells (SSCs) is provided. The instant disclosure encompasses the discovery and optimization of distinct, stage-specific culture media and conditions that enable, for the first time, the complete progression of human spermatogenesis outside the body. The process includes: (1) inducing entry of dSPGs and SSCs into meiosis; (2) supporting meiotic progression and completion to round spermatids; and (3) promoting spermiogenesis and the formation of elongated, spermatozoa. The invention further provides compositions, systems, and protocols for each stage, as well as markers and methods for identifying successful progression through each developmental transition. This system enables, for the first time, the complete in vitro recapitulation of human spermatogenesis, providing a platform for research, infertility treatment, and assisted reproductive technologies.

The utility model belongs to the technical field of cell culture, and particularly relates to an automatic cell collecting device which comprises a rack and a sealing cover arranged on the rack, a normally closed butt joint window is arranged on one side of the sealing cover, and the butt joint window can be connected with a culture box; a culture dish can be placed in the culture box; the first mechanical arm is arranged on the operation platform; the liquid collecting mechanism is arranged on the operation platform, the liquid input mechanism is arranged on the operation platform, a first cover opening assembly is further arranged beside the liquid input mechanism, and the centrifugal collecting mechanism is arranged on the operation platform. By arranging the butt joint window, the first mechanical arm, the liquid collection mechanism, the liquid input mechanism and the centrifugal collection mechanism, the cell collection process can be fully automatically completed, the automation degree is high, the risk of artificial secondary pollution is reduced, meanwhile, a production line can be synthesized with a cell inoculation device, the first mechanical arm is shared, the size of the machine is reduced, and the production cost is reduced. The occupied area is small, the cost is low, and the structure is simpler.

The present application relates to the technical field of biological cell culture, and particularly relates to a three-dimensional cell culture device with automatic liquid replacement function, comprising: a box body; a rotating assembly, a liquid replacement assembly and a control display assembly are integrally installed in the box body. The three-dimensional cell culture device with automatic liquid replacement function provided by the present application integrates microgravity simulation and liquid replacement function in a relatively closed culture box body space, ensures that the culture environment in the cell culture process has high stability by not excessively affecting the air flow, temperature and humidity environment, etc. in the culture box body during liquid replacement; the device for replacing culture solution does not need to rotate, effectively releases the culture space while reducing the energy consumption required in the rotation process, can load multiple culture bottles for three-dimensional culture and automatic liquid replacement at the same time, and has high potential for large-scale cell preparation.

The invention discloses a method for culturing NK (Natural Killer) cells. The method comprises the following steps: inoculating a mononuclear cell into a T25 cell culture flask, wherein the T25 cell culture flask is coated by a coating factor in advance; carrying out culture treatment on the inoculated mononuclear cells in a culture system containing an NK cell culture medium containing an activation amplification factor and plasma so as to obtain NK cells; wherein the coating factor, the activation amplification factor and the NK cell culture medium are derived from an NK cell amplification kit. According to the method provided by the embodiment of the invention, the dosage of the amplification reagent can be reduced, the amplification multiple of the NK cells is improved, and the production cost is reduced.

This invention discloses a hepatocyte model with silencing or overexpression of LCMT1 and its construction method, comprising: passage and seeding cells in logarithmic growth phase into well plates; diluting plasmid siRNA1 or siRNA2 or an overexpression plasmid to an appropriate concentration, and then mixing it with transfection reagent dilution to obtain a transfection mixture; washing the cells in each well of the plate and adding another culture medium, then gently dripping the transfection mixture into the well plate, shaking well, and culturing in a cell culture incubator for a period of time; replacing the culture medium with complete culture medium without antibiotics to obtain the constructed hepatocyte model with silencing or overexpression of LCMT1. This invention, by constructing a hepatocyte model with silencing or overexpression of LCMT1 in the liver, allows for validation experiments to be conducted to study the development mechanism of liver fibrosis. It is suitable for widespread application in various laboratories and provides an experimental model basis for research on liver fibrosis and drug trials.

The invention relates to biomedical engineering, and discloses a biochip screening method of a therapeutic agent based on intestinal brain axis regulation and application thereof. The screening method comprises the following steps: performing intestinal wall cell culture or microglial cell culture by adopting a metasurface plasmon resonance biochip of which the surface is subjected to bionic treatment to obtain an intestinal wall cell adherent chip board or a microglial cell adherent chip board; the method comprises the following steps: adding a culture medium containing a chemotherapeutic drug or lipopolysaccharide into a cell-adherent chip board for culture I, then removing the culture medium, adding a therapeutic agent to be detected for culture II to obtain a culture solution, and detecting the cell repair effect of intestinal wall cells or microglial cells in the culture solution, and screening to obtain a therapeutic agent with a repairing effect on intestinal wall cells and / or microglial cells. The screening method can be used for efficiently and accurately screening the therapeutic agent with dual effects of intestinal regulation and neuroprotection from a plurality of therapeutic agents to be detected, and can be used for dynamically monitoring and exploring the interaction condition of the therapeutic agent and cells on line in an unmarked and real-time manner.