235 results about "Culture cell" patented technology

The invention discloses a preparation method and application of a cell culture liver based on a microcarrier. The method comprises the steps of obtaining liver seed cells, preparing the microcarrier, culturing cells in vitro based on the microcarrier, preparing the cell culture liver and the like. The preparation method of the cell culture liver based on the microcarrier is provided for the first time, and the preparation scheme that a cell-microcarrier compound serves as a structural unit and is assembled into the cell culture liver after being mixed with an edible material is adopted, so that on one hand, simulation of a real liver lobule structure can be achieved, and on the other hand, simulation of the liver lobule structure can be achieved; the whole structure relation of liver lobules and connective tissues in liver tissues can be simulated, full reduction of animal liver appearance and structural characteristics is achieved, edible materials used in the preparation process are low in price and good in biocompatibility, and the preparation method is suitable for large-scale industrial production. Liver seed cells can be subjected to efficient amplification and albumin synthesis through three-dimensional culture on a microcarrier prepared on the basis of edible materials, so that the cell culture liver is close to a real animal liver in the aspect of nutrition.

The invention discloses a perfusion device for stem cell culture, and belongs to the technical field of cell culture.The perfusion device comprises a frame, a closing assembly convenient to overhaul is arranged on the frame, a culture container is arranged in the frame, a perfusion bottle is fixedly installed in the frame, a filter box is fixedly installed in the frame, and the filter box is connected with the perfusion bottle. A pumping assembly is arranged in the frame, a penetrating groove is formed in the top of the filter box, and a connecting pipe is connected to the filter box; culture cells are conveniently collected through the arranged filter cylinder, the cells cultured in the perfusion bottle and a culture solution flow into the filter box through a liquid supply pipe, the filter cylinder is covered with a filter membrane, the liquid can flow to the outside to intercept the cells, then the cells can be filtered in the filter cylinder to be collected, then the filter cylinder is rotated, and the cells can be conveniently collected. The threaded sleeve is separated from the threaded head, so that the filter cartridge can be detached, the cells can be conveniently separated and collected, and the working efficiency is improved.

The present invention relates to a microfluidic device (10) for 3D cell and tissue culture, wherein said microfluidic device (10) comprises at least one microfluidic unit (11) comprise a scaffold chamber (14) for housing a matrix (19) for culturing cells and / or acting as a barrier, in fluid communication with a media chamber in the form of a loop chamber (12) having a hollow section (16) for guiding a circular flow path of a fluid therein and a first capillary burst valve (15a) positioned between said scaffold chamber (14) and said loop chamber (12) arranged to selectively control the flow of fluid therebetween.

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 invention provides polypeptides and encoding nucleic acids of aldehyde dehydrogenase variants. The invention also provides cells expressing aldehyde dehydrogenase variants. The invention further provides methods for producing 3-hydroxybutyraldehyde (3-HBal) and / or 1,3-butanediol (1,3-BDO), or an ester or amide thereof, comprising culturing cells expressing an aldehyde dehydrogenase variant or using lysates of such cells. The invention additional provides methods for producing 4-hydroxybutyraldehyde (4-HBal) and / or 1,4-butanediol (1,4-BDO), or an ester or amide thereof, comprising culturing cells expressing an aldehyde dehydrogenase variant or using lysates of such cells.

In one embodiment, cells 12 are suspended inside a hollow capsule 11 including a semipermeable membrane, and the hollow capsule 11 is incubated in a culture medium to allow the cells 12 to aggregate inside the hollow capsule 11. The cells 12 are a mixture of dissociated target cells 14 and auxiliary cells 15 for assisting the aggregation of the target cells 14. The target cells 14 more easily aggregate in the presence of the auxiliary cells 15 than in the absence of the auxiliary cells 15. In another embodiment, dissociated cells 12 are suspended inside a hollow capsule 11 including a semipermeable membrane, and the hollow capsule 11 is incubated in a culture medium to allow the cells to aggregate inside the hollow capsule 11. The cells 12 include at least stromal cells. Thus, an aggregate 21 of the cells 12 is produced.

The invention relates to the technical field of cell culture, in particular to an oriented porous bionic gelatin scaffold for controlling cell orientation arrangement, a preparation method and application thereof, and the oriented porous bionic gelatin scaffold can be used for cell culture meat production. The preparation method comprises the following steps: mixing and solidifying gelatin and glutamine transaminase, directionally freezing, freeze-drying, sterilizing, inoculating and culturing cells and the like. Parameters and culture medium components in each step are also explained. Compared with the prior art, a regular directional porous structure is formed through a hydrogel scaffold and directional freezing, directional arrangement and growth of fish muscle satellite cells are guided, the cell activity is high, and the proliferation and differentiation effects are good; the gelatin material is adopted, so that the cost is low, the biocompatibility is good, and the gelatin material is superior to collagen and wheat glutelin; the preparation process is simple, does not need complex equipment, is suitable for large-scale production, and promotes the development of cell culture meat.

The invention discloses a cfDNA detection standard substance and a preparation method thereof. The preparation method comprises the following steps: carrying out multiplication culture on cells; performing cell apoptosis treatment on the amplified cells; after cell apoptosis treatment is finished, incubating for a period of time, and collecting supernate; adding the supernate into plasma to prepare a plasma standard substance for cfDNA detection; or extracting cfDNA in the supernate, and preparing a DNA standard substance for cfDNA detection; according to the preparation method, the supernatant of a culture cell line is adopted as a source of a cfDNA detection standard substance, rapid death of cells is promoted through a method of nutrient depletion or cytotoxic reagent addition, the content of mononuclear bodies in the supernatant of the culture medium is increased, the concentration of cfDNA is further increased, and the preparation method is easy to operate, high in yield and low in cost.

The invention particularly relates to a chip co-culture method, a high-content imaging analysis method based on the chip co-culture method and application of the chip co-culture method in anti-tumor drug screening. The tumor-related fibroblasts and the tumor organoid cells are respectively subjected to resuspension counting with a PBS buffer solution, the two kinds of cells are fully and uniformly mixed according to the quantity ratio of 1: (2-3) to form co-culture cell suspension, and the co-culture cell suspension is inoculated into a chip according to the volume dose of 1.5-2 mL per row; placing the culture chip in a 5% CO2 incubator at 37 DEG C for first culture, after incubation for 20-30 minutes, slightly and completely sucking PBS in the chip by using a pipette, adding 1.5-2 mL of an organoid culture medium into each hole of the chip, and placing the chip in the 5% CO2 incubator at 37 DEG C for second culture to obtain the co-culture cell system. Rapid and high-quality detection of a large amount of samples can be realized, and an important technical support is provided for deeply understanding a tumor microenvironment and developing a new treatment strategy.

The invention relates to a co-culture microsphere model with a core-shell structure and a preparation method of the co-culture microsphere model. The co-culture microsphere model comprises a core-shell microsphere, the core-shell microsphere is composed of an inner core and an outer shell, the inner core is an organoid loaded by matrigel, the outer shell is formed by crosslinking a photo-crosslinking hydrogel material, co-culture cells are doped in the photo-crosslinking hydrogel material, and the inner core is wrapped by the outer shell. According to the co-culture microsphere model, double-layer structure microspheres of co-culture cell wrapped organoids are formed, immune infiltration and in-vivo characteristics can be simulated, and a similar in-vivo three-dimensional tumor immune microenvironment is provided. And the types of cells loaded on the inner layer and the outer layer can be flexibly replaced to form a microsphere model for co-culture of single or multiple cells and organoids. The co-culture microsphere model is more stable in structure, the size of the diameter of the core-shell microsphere can be optimized by changing 3D printing parameters, the operability of subsequent research is high, and the co-culture microsphere model is suitable for different research application scenes.

The present invention relates to a polymer conjugate comprising a ULA polymer a) and one or more polypeptides b), wherein the polypeptides are conjugated to the ULA polymer, and the conjugate is coated on the surface of a device for culturing cells. The peptides can interact with receptors on the surface of the cells, while the polymer prevents non-specific binding of the cells to the surface of the device.

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

The present invention relates to a method for culturing cells, as well as a cell culture device and a kit used in the culturing method. The method for culturing cells of the present invention includes applying cells to a polyimide porous membrane and culturing them. In one embodiment of the method of the present invention, it includes a step of inoculating cells onto the surface of the above-mentioned polyimide porous membrane. Alternatively, in one embodiment, it includes the following steps: loading a cell suspension onto the dry surface of the above-mentioned polyimide porous membrane, placing the above-mentioned polyimide porous membrane, or moving the above-mentioned polyimide porous membrane to promote the outflow of liquid, or stimulating a part of the surface to suck the cell suspension into the above-mentioned membrane, and then accumulating the cells in the cell suspension into the above-mentioned membrane and allowing the water to flow out. Alternatively, in one embodiment, it includes the following steps: wetting one or both surfaces of the above-mentioned polyimide porous membrane with a cell culture solution or a sterilized liquid, loading a cell suspension onto the wetted polyimide porous membrane, and then accumulating the cells in the cell suspension into the above-mentioned membrane and allowing the water to flow out.

The invention belongs to the field of veterinary virology, and discloses a bovine ephemeral fever virus strain and reverse genetic application thereof, and the reverse genetic steps are as follows: taking pCI plasmid as a skeleton to prepare plasmid for expressing a complete genome of bovine ephemeral fever virus; the method comprises the following steps: respectively preparing auxiliary plasmids for expressing an N protein gene, a P protein gene, an L protein gene and a G protein gene of the bovine ephemeral fever virus by taking pCAGGS plasmids as a skeleton, co-transfecting BHK-21 cells by using the five plasmids, and culturing the cells to obtain the rescued bovine ephemeral fever virus. The rescued bovine ephemeral fever virus has no obvious difference from the reproductive capacity of the parent virus, and the genome is still stable after 10 generations of continuous passage in BHK-21 cells. The invention provides methodological support for genetic modification of the bovine ephemeral fever virus, and has an application prospect.

The present invention relates to a method for three-dimensional culture and induced differentiation of neural organoids and use thereof. The method for three-dimensional culture and induced differentiation of neural organoids comprises: adding a signal pathway inhibitor into an induced pluripotent stem cell three-dimensional culture, and culturing cells to obtain the neural organoids. The signal pathway inhibitor comprises a TGF-β signal pathway inhibitor, a WNT signal pathway inhibitor, and a BMP signal pathway inhibitor. Compared with a traditional method, the neural organoids obtained by the method of the present invention feature a large size, small individual difference, and short culture time. The method is simple and rapid, and has high stability and low cost, and only a common incubator is needed. The method has wide application prospects.

To provide a culture vessel by which cells can be easily observed.SOLUTION: In a method for manufacturing a culture vessel, a culture vessel having a body part which has a through hole, and a sheet stuck on a lower face of the body part so as to close a lower end part of the through hole is manufactured. The method includes the steps of: adding tensile force to a sheet material with a thickness dimension of less than 50 μm; obtaining a first intermediate body in which the sheet material is stuck on a substrate; and sticking a surface of the sheet material side of the first intermediate body to the lower face of the body part.SELECTED DRAWING: Figure 2

This product is a cell manufacturing device that processes and cultures cells by setting a cartridge containing blood and reagents into the main body.

An object of the present invention is to provide a cell culture method and a production method for a product, in which an occurrence of aggregates in foam is suppressed.According to the present invention, there is provided a cell culture method of culturing cells at a cell density of 30×106 cells / mL or more and 400×106 cells / mL or less in a culture solution, the method including: step A of adjusting a poloxamer concentration in a foam liquid constituting foam to be 6 times or less a poloxamer concentration in the culture solution; or step B of adjusting a viscosity of the foam liquid constituting the foam to be 1.5 mPa·s or less under conditions of 35° C. or higher and 38° C. or lower.

The invention relates to a pueraria flower quality control and evaluation method based on reactive oxygen species (ROS) detection, and can effectively solve the problem that an existing pueraria flower quality detection method is difficult to reflect the overall efficacy of pueraria flowers. The method comprises the following steps: preparing a pueraria flower alcohol extract, constructing an alcohol damage cell model, preparing a cell culture medium containing gradient pueraria flower extract, culturing cells, and carrying out ROS level detection and statistics. The invention relates to a control group, a model group and a kudzuvine flower extract group, the internal quality of the control group, the model group and the kudzuvine flower extract group are jointly controlled from multiple angles, the current quality control and evaluation method is improved, a new technical approach can be provided for quality control and quality evaluation research of other alcohol effect dispelling and liver protecting traditional Chinese medicine decoction pieces, and the method is easy to operate, high in practicability, wide in application range and easy to popularize. The method is an innovation in traditional Chinese medicine quality control and evaluation.

The invention provides polypeptides and encoding nucleic acids of aldehyde dehydrogenase variants. The invention also provides cells expressing aldehyde dehydrogenase variants. The invention further provides methods for producing 3-hydroxybutyraldehyde (3-HBal) and / or 1,3-butanediol (1,3-BDO), or an ester or amide thereof, comprising culturing cells expressing an aldehyde dehydrogenase variant or using lysates of such cells. The invention additional provides methods for producing 4-hydroxybutyraldehyde (4-HBal) and / or 1,4-butanediol (1,4-BDO), or an ester or amide thereof, comprising culturing cells expressing an aldehyde dehydrogenase variant or using lysates of such cells.

The present invention provides an inbred corn line designated JRDJ6933, methods for producing a corn plant by crossing plants of the inbred line JRDJ6933 with plants of another corn plant. The invention further encompasses all parts of inbred corn line JRDJ6933, including culturable cells. Additionally provided herein are methods for introducing transgenes into inbred corn line JRDJ6933, and plants produced according to these methods.

One approach to the exploration method includes calculating a TAD score representing transcriptional activity for each TAD present in the genome and selecting TADs based on the TAD scores. Another approach includes analyzing a genome obtained from cells expressing a foreign gene and selecting the boundaries of regions containing the foreign gene. Cell preparation involves inserting the target gene into the regions identified by the exploration method. Cell sorting involves calculating a TAD score representing transcriptional activity for each TAD present in the genome, selecting TADs based on the TAD scores, and selecting cells containing the target gene in the selected TADs. The cells are derived from Chinese hamsters and are cells in which the target gene has been inserted into at least one specified region. The method for manufacturing the cell product includes culturing the cells to express the target gene.

To provide safe and high-performance stem cells appropriate for clinical application, and a method for producing the stem cells.SOLUTION: Provided are a stem cell population derived from human deciduous tooth dental pulp, wherein 90% or more of the stem cell population is characterized by being CD117-negative, CD73-positive, CD90-positive, and CD105-positive, and a method for producing a stem cell population derived from human deciduous tooth dental pulp, the method comprising a step of culturing cells isolated from human deciduous tooth dental pulp in a medium that does not contain FBS (fetal bovine serum) in the presence of human platelet lysate (hPL).SELECTED DRAWING: None

Methods are disclosed for producing a polypeptide comprising an amino acid sequence including a lysine residue susceptible to hydroxylation, and / or including a proline residue susceptible to hydroxylation, comprising culturing cells comprising nucleic acid for expressing the polypeptide for the majority of the period of culture in cell culture medium having an Fe ion concentration of less than 60 µM.

The application provides a method for preparing FFPE quality control products by using cell lines, a reference product and a kit, and belongs to the fields of medicine, clinical laboratory science and biotechnology, and comprises the following steps: culturing cell lines by using microcarriers; mixing different cell lines based on requirements; centrifuging the mixed cell microcarrier suspension in a sealed malt pipe at one end; removing supernatant after the treatment; packaging the two ends of the malt pipe by using a semi-permeable membrane; removing the semi-permeable membrane after fixing, dehydrating and transparentizing the malt pipe by using a formaldehyde solution; separating the cell microcarriers after the wax immersion treatment from the malt pipe; and embedding the cell microcarriers into a paraffin block by using wax. The quality control product can be used in subsequent quality control processes, the mutation frequency of the quality control product can be kept in good uniformity, and batch production and large-scale production of the quality control product can be realized.

An apparatus for culturing cells includes a bioreactor with a fluid reservoir and a fixed bed, as well as a portion associated with the fluid reservoir and adapted for preventing cells from settling therein. The bioreactor may include a first vessel including the fluid reservoir and a second vessel with the fixed bed for the cell culture, which may be directly connected to the first vessel by a frangible connection to facilitate disassembly, such as for sampling. A conduit may be provided for connecting the first vessel to the second vessel, so as to establish fluid communication there between. A pump may also be provided for pumping fluid from the first vessel to the second vessel, such as though the conduit if present. Use of the apparatus as a screening tool for a large-scale bioreactor, in a kit, and related methods are also disclosed.

The present invention provides an inbred corn line designated KFF7351, methods for producing a corn plant by crossing plants of the inbred line KFF7351 with plants of another corn plant. The invention further encompasses all parts of inbred corn line KFF7351, including culturable cells. Additionally provided herein are methods for introducing transgenes into inbred corn line KFF7351, and plants produced according to these methods.

The invention belongs to the technical field of cell biology, and particularly relates to a cell culture method based on self-assembly microgel, a cell product and application. The cell culture method based on the self-assembled microgel comprises the following steps: culturing cells by adopting a cell culture medium, and adding the self-assembled microgel into a culture system. The cell culture method has the beneficial effects that the controllable and reversible self-assembly microgel scaffold is constructed by taking microgel with self-assembly capability as a basic construction unit, and three-dimensional wrapping, static culture and dynamic culture aiming at suspension cell types are realized in vitro; the cells obtained by the culture method can be transplanted in vivo, can be used for a clinical treatment scheme for assisting transplantation of cells in marrow cavities such as allogeneic hematopoietic stem cells and megakaryocytes, and can be used for optimizing cell therapy of hematological malignant tumors, thrombocytopenia and other hematological system malignant diseases.

The utility model relates to the technical field of cell culture, and discloses a cell shake flask capable of improving tissue digestion efficiency, which comprises a culture flask body, a flask cover is mounted at the top end of the culture flask body, and a first constant-temperature heating cover is arranged on one side of the outer side wall of the culture flask body. Mounting blocks are fixed to the front end and the rear end of one side of the first constant-temperature heating cover correspondingly, inner cavities are formed in the first constant-temperature heating cover and the second constant-temperature heating cover correspondingly, and electric heating wires are mounted in the inner cavities. When the culture bottle body disclosed by the utility model is used for culturing cells, the culture bottle body can be heated according to actual conditions by arranging components such as the first constant-temperature heating cover, the mounting block, the mounting groove and the second constant-temperature heating cover, and meanwhile, the digestion efficiency of cell tissues can be promoted by heating the culture bottle body; and the culture bottle body can be detached when not needed subsequently, so that the observation of the condition of cells in the culture bottle body through the culture bottle body is not influenced, and the use effect is better.

Provided is a method for producing, by culturing, a cell for measuring the activity of a sensor protein that can be used as a chemical substance sensor. A method for producing a cell for measuring the activity of a sensor protein, the method comprising: culturing a cell containing a polynucleotide containing a sensor protein coding sequence, and when the ratio (specific activity) of the activity value of the sensor protein when the concentration of a response substance is 1 [mu] M to the activity value when the concentration of the response substance is 0 [mu] M is 1.2 or more; and recovering the cells.