41 results about "Oxyntic cell" patented technology

A nanostructured low-immunogenic biological artificial blood vessel, a preparation method therefor and a use thereof. The preparation method comprises: decellularizing a pre-treated animal blood vessel to obtain a decellularized blood vessel; treating the decellularized blood vessel in an enzyme solution to obtain an enzyme-treated blood vessel, wherein the enzyme solution comprises nuclease and / or a biological enzyme; and using a cross-linking agent to cross-link the enzyme-treated blood vessel to obtain a nanostructured low-immunogenic biological artificial blood vessel, which is used as a blood vessel transplantation material. The artificial blood vessel overcomes the defects of decreased mechanical properties in decellularized biological tissues, in-vivo calcification after long-term use, and the presence of immunogenicity, antigenic components such as vascular wall cells and cell nuclei are fully removed, and original collagen and other extracellular matrix structural proteins in the tissue are retained to a greater extent, avoiding the in-vivo calcification of the blood vessel upon long-term implantation, enhancing the durability of use of the blood vessel; and the method has a short preparation cycle, low cost and high long-term patency rate.

The invention relates to the field of cell culture equipment, in particular to an adherent cell culture reactor, and aims to solve the problems that some cells at the bottom of a culture dish are easy to lack nutrition when the cells are subjected to static culture at present, so that growth stagnation and even necrosis are caused; according to the adherent cell culture reactor provided by the invention, the problem that the cells are easily damaged and even die due to shearing force generated by stirring a culture solution through stirring equipment is solved, the adherent cell culture reactor is provided, and the adherent cell culture reactor consists of a left shell and a right shell which are symmetrically arranged and side plates which are fixedly connected between the two shells and are arranged on the front side and the rear side; the lower side of the vertical plate is provided with a driving structure for controlling the incubator to rotate at a steady speed, the inner walls of the two protective covers are respectively and fixedly connected with a culture bed, and the adherent cells can be driven to uniformly stay in the culture beds in the steady-speed rotation process of the incubator; the culture solution flows towards one direction in the steady-speed rotation process of the culture device, so that the shearing force generated during the circulation of the culture solution can be greatly reduced, and the damage to cells is reduced.

The invention belongs to the technical field of medicines, and particularly provides a solid dispersion as well as a preparation method and application thereof. The solid dispersion comprises a carrier material, and further comprises quercetagetin and phospholipid, the mass ratio of the quercetagetin to the phospholipid is 1: (0.3-1.5), and the carrier material comprises one or more of a hydrophilic polymer and a pH-dependent polymer. Wherein the quercetagetin and the phospholipid are combined through an intermolecular force, so that the quercetagetin obtains a lipid material similar to a cell membrane structure, a cell membrane bionic phospholipid compound is formed, the cell membrane bionic phospholipid compound can be naturally compatible with an upper wall cell membrane of a small intestine, and the permeation transmembrane capability of the quercetagetin is better promoted; the quercetagetin is added into the solid dispersion, so that more quercetagetin can enter systemic circulation and lymphatic circulation, and therefore, the solid dispersion can strengthen transmembrane absorption while strengthening dissolution, so that the solid dispersion has high bioavailability.

To provide methods of producing and purifying AAV viral vectors having low empty capsids, low host cell proteins, and / or low contaminating DNA, while retaining high potency; and to provide a pharmaceutical composition containing the vector.SOLUTION: A method of manufacturing an AAV viral vector comprises: a. culturing adherent cells; b. transfecting the adherent cells with one or more plasmids to enable production of the AAV viral vector; c. lysing the adherent cells to isolate the AAV viral vector; d. acidifying and clarifying the cell lysate of c; e. purifying the product of d using cation exchange chromatography; f. filtering the product of e using tangential flow filtration; g. ultracentrifuging the product of f in a cesium chloride buffer; h. collecting the AAV viral vector from the product of g; and i. formulating and filling the AAV viral vector collected in h. to produce a final pharmaceutical product.SELECTED DRAWING: None

The invention discloses a method for realizing in-vitro amplification of adherent cells by utilizing a stirring type bioreactor matched with a three-dimensional porous hydrogel carrier. The method comprises the following steps: sterilizing a hydrogel carrier and a stirred bioreactor, adding a cell culture medium, ventilating, inoculating cells, starting the stirred bioreactor for culturing, observing the color change of a culture solution in the culture process, and changing the solution when the culture solution turns yellow, thereby ensuring the nutrition required by cell culture. The hydrogel with high biocompatibility is used as a carrier, a porous three-dimensional growth environment is provided, cells can be attached to the interior of the hydrogel carrier, the cell growth space is sufficient, the cells can be cultured for more generations, the number and density are greatly increased, the amplification multiple can reach 10 times or above, the production efficiency is improved, and the production cost is reduced.

The mechanisms driving the development of extracapillary lesions in focal segmental glomerulosclerosis (FSGS) and crescentic glomerulonephritis (CGN) remain poorly understood. A key question is how parietal epithelial cells (PECs) invade glomerular capillaries, thereby promoting injury and kidney failure. Here the inventors show that expression of the tetraspanin CD9 increases markedly in PECs in mouse models of CGN and FSGS, and in kidneys from individuals diagnosed with these diseases. Cd9 gene targeting in PECs prevents glomerular damage in CGN and FSGS mouse models. Mechanistically, CD9 deficiency prevents the oriented migration of PECs into the glomerular tuft and their acquisition of CD44 and β1 integrin expression. These findings highlight a critical role for de novo expression of CD9 as a common pathogenic switch driving the PEC phenotype in CGN and FSGS, while offering a potential therapeutic avenue to treat these conditions. Accordingly, CD9 represents a reliable biomarker and as well as a biotargets in glomerulonephritides.

The invention discloses a multilayer adherent cell culture reactor and a use method. The culture reactor comprises a shell, a plurality of layers of cell culture plates are arranged in the shell in an array manner; and the liquid inlet is used as an inlet of a cell suspension. The liquid outlet is an outlet of the culture medium; the air holes are formed in the upper surface and the lower surface of the shell. And the quality control chamber is communicated with the inner cavity of the shell. The use method comprises the following steps: uniformly mixed cells and a culture medium enter the inner cavity of the shell through the liquid inlet for positive culture; after front culture is completed, the multi-layer adherent cell culture device is inverted; uniformly mixed cells and a culture medium enter the inner cavity of the shell through the liquid inlet for reverse culture. By arranging the multiple layers of cell culture plates with double surfaces capable of culturing, the cell culture area is greatly increased, the space utilization rate is improved, the use amount of a culture medium is reduced, the prepared quality control chamber is communicated with the inner cavity of the shell, and the cell growth state is conveniently, rapidly and accurately observed on the premise of not polluting the internal environment and not damaging the culture stability through isolation of the partition opening.

The present invention discloses a novel uniform strain chamber for an adherent cell loading device, which relates to the field of cell mechanics. Cells are planted in the novel uniform strain chamber of the adherent cell loading device, and the four positioning holes of the chamber are placed on a fixed pile. The fixed pile is connected to the stator. The controller controls the stepper motor, and the rotational motion of the motor is converted into linear motion through a transmission screw and transmitted to the mover, thereby indirectly strain stimulating the cells in the chamber cavity, thereby realizing relevant mechanical biology research on the cells. The strain of the adherent cell chamber in the existing cell tensile loading device is uneven, and the experimental results have large errors. In the invention, the elastic basement membrane of the chamber is designed to have one side as a plane, i.e., the part where the planted cells are poured into the cell culture medium, and the other side as a curved surface, i.e., an optimized surface. The longitudinal cross-section is an M-shaped structure, and its thickness variation meets the uniformity requirement of the strain of the plane part of the basement membrane during the chamber loading process, thereby increasing the uniform area of ​​the adherent cells in the strain loading, and therefore has high use value.

The utility model discloses an adherent cell culture medium, which belongs to the technical field of culture media for adherent cells and comprises a shell, a partition plate arranged in the shell, four cavities arranged between the partition plate and the shell, a culture dish arranged in each cavity, and a solid culture medium arranged at the bottom of the inner side of each culture dish. A pop-up assembly is arranged between the culture dish and the shell, the pop-up assembly is used for popping up the culture dish, the pop-up assembly comprises a threaded rod, the threaded rod is arranged on the lower side of the culture dish, the lower side of the shell is rotationally connected with a nut, the nut is in threaded connection with the threaded rod, and a plurality of springs are arranged between the culture dish and the shell; and an adjustable shading component is arranged on the upper side of the shell and is used for performing different shading adjustments on the culture dishes in the cavity, so that the problem that a plurality of groups of adherent cells are inconvenient to culture and observe by using the same culture medium is solved.

The disclosure is directed to a method for seed train expansion of adherent cells comprising culturing cells with a serum-supplemented growth medium in a N-2 vessel; removing the cells from the serum-supplemented medium; inoculating the cells from step into a serum-free growth medium in a N-1 vessel; culturing the cells in the N-1 vessel under suspension conditions; and inoculating a growth medium in a bioreactor with the suspension-cultured cells. In some aspects, the adherent cells are not suspension-adapted. In some aspects, the adherent cells are suspension-adapted. In some aspects, the adherent cells produced by the seed train expansion method are used to produce viral vectors. In some aspects, the viral vectors are AAV vectors.

The invention relates to the technical field of biological medicine, and discloses a natural product for targeted inhibition of a KCNQ1 / KCNE2 complex channel and application of the natural product for targeted inhibition of the KCNQ1 / KCNE2 complex channel in preparation of a medicine for treating or preventing gastric ulcer, and an application method of a pharmaceutical composition comprises the following steps: S1, applying the pharmaceutical composition to a medication object; s2, the natural product interacts with a KCNQ1 / KCNE2 complex channel on a gastric wall cell membrane, and potassium ion current of the complex channel is inhibited; s3, weakening functions of H < + > / K < + >-ATPase in gastric wall cells by inhibiting potassium ion current, so that gastric acid secretion is reduced, and gastric ulcer is treated or prevented. According to the invention, gastric acid is indirectly regulated and controlled through targeted inhibition of gastric wall cell KCNQ1 / KCNE2 complex channels, and a new strategy for treating gastric ulcer, which is milder and has safety potential, is provided.

The invention relates to a cell culture device (10, 10a, 10b) comprising - a culture chamber (15, 15a; 15b) comprising a cell culture medium (25), - a plurality of cell culture supports (30, 30a; 30b) present in the cell culture medium (25) contained in the culture chamber (15, 15a; 15b), the cell culture supports (30, 30a; 30b) having a density different from that of the cell culture medium (25) such that these cell culture supports (30, 30a; 30b) are located at an interface (27) at least constituted on one side by the cell culture medium (25), at least one of the cell culture supports (30, 30a; 30b) carrying adherent cells (C), and - a system for moving the cell culture supports (30, 30a; 30b) along the interface (27).

The present application relates to the technical field of biology, and particularly discloses a method for culturing and obtaining foregut endoderm cell spheroids and application thereof. The method for culturing and obtaining foregut endoderm cell spheroids of the present application obtains foregut endoderm cell spheroids by inducing differentiation of stem cells on a culture surface with a PDA-matrigel coating. The culture method of the present application can ensure the differentiation efficiency of DE and AFE adherent cells and spheroids, promote the generation of AFE cell spheroids and the separation from the adherent cell layer, reduce the edge rolling of AFE adherent cells, prolong the production time of AFE cell spheroids, significantly improve the yield of AFE cell spheroids, and provide a basis for subsequent differentiation and culture of lung organoids.

The invention discloses a bioreactor for nerve cell culture, and relates to the technical field of cell culture, a metal ball falls down to drive a culture dish to deflect, so that an azimuth magnetic field is generated, multi-stage energy storage is performed on the metal ball, the metal ball is bounced off after the energy is stored to a certain degree, and the bounced-off metal ball falls down disorderly to drive the culture dish to deflect disorderly. Therefore, the magnetic field is generated disorderly, irregular magnetic field stimulation is generated on nerve cells in the culture dish, and on one hand, the basic problems of cell adherence, cell block mass precipitation, uniform distribution of nutrient substances and oxygen, removal of metabolic waste and the like are solved; on the other hand, a stirring structure is not arranged, so that the problem of damage to cells caused by traditional stirring is avoided; and finally, the nerve cells are stimulated by combining a magnetic field disordered triggering mode with disordered deflection of the culture dish, so that the initiative of cell movement such as nerve cell axon growth, synaptic remodeling, cell migration and the like is stimulated.

The invention provides a method for obtaining patterned adherent cells, which comprises the following steps of: covering the surfaces of the adherent cells with a solution containing photo-initiated photosensitive hydrogel, carrying out patterning treatment on an obtained hydrogel system through stereolithography, and removing an uncured hydrogel part and cells which are not protected by the cured hydrogel to obtain the patterned adherent cells. And finally, removing the cured hydrogel to obtain the adherent cells subjected to patterned culture. According to the method, the sacrificial phase hydrogel protective layer is manufactured in combination with a photoetching method, the adherent cells do not need to be resuspended again, the cell pollution risk possibly introduced in a traditional method is avoided, in-situ arrangement and shape control of the adherent cells can be achieved, and adherent cell communities with regular or complex patterns can be formed.

The present invention discloses an evaluation method for adherent cell digestion results, a cell culture method and a terminal device. The evaluation method for adherent cell digestion results comprises: obtaining an adherent cell image after digestion; performing image processing on the adherent cell image after digestion to obtain the adherent cell area after digestion; calculating the digestion yield according to the adherent cell area after digestion and the bottom area of ​​the cell culture container; or calculating the digestion yield according to the adherent cell area after digestion and the adherent cell area before digestion; evaluating the adherent cell digestion results according to the digestion yield to obtain a digestion evaluation result. The present invention can avoid the error caused by manually evaluating the digestion results of adherent cells, so as to improve the accuracy of the digestion evaluation results of adherent cells, thereby ensuring the cell viability of adherent cells during the culture process.

To provide a substrate for suspension culture of adhesive cells, which is used for suspension culture of adhesive cells capable of producing a substance selected from proteins, glycoproteins, viruses, and viral vectors, wherein the substrate exhibits superior culture performance and enables further increase in the production of the substance.SOLUTION: A substrate for suspension culture of adhesive cells comprises a laminate in which a porous layer and a nonwoven fabric layer having an apparent density of less than 0.440 g / cm3 are laminated.SELECTED DRAWING: None

An adherent cell culture substrate, which is a sheet-shaped substrate used for producing a culture vessel for adherent cells, includes a groove including a crest-shaped portion and a valley-shaped portion on one surface side of the substrate, in which a distance from a top end portion of the crest-shaped portion to the other surface of the substrate is 1 mm or less. Further, it is preferable that a cross section of the groove perpendicular to a direction in which the groove extends is formed in a substantially V-shape and an inclination angle of a side surface of the substantially V-shape of the groove is 80 degrees or less. Further, it is preferable that a plurality of the grooves are linearly arranged in parallel and the top end portions of the crest-shaped portions are formed in a linear shape.

The application belongs to the technical field of animal models, and particularly relates to a construction method and application of a GRIM-19 and IL-33 double-gene parietal cell-specific knockout mouse model. The construction method comprises the following steps: respectively modifying mouse GRIM-19 and IL-33 genes by flox, then crossing the GRIM-19flox / flox mouse with the IL-33flox / flox mouse to obtain F1 generation mice; crossing the F1 generation mice with Atp4b-Cre mice to obtain F2 generation mice; crossing the F2 generation mice with the F1 generation mice to obtain a GRIM-19 and IL-33 double-gene parietal cell-specific knockout mouse model. The model is helpful for exploring the regulation mechanism of the GRIM-19 and IL-33 genes in SPEM occurrence and progression, and is of great significance for the treatment of SPEM and the prevention and treatment of gastric cancer.

The invention relates to an adherent cell microcarrier suspension culture method, which can reduce the usage amount of serum, reduce the cost of a culture medium and the risk of biosafety, can flexibly enlarge the culture volume according to the specification of a shake flask, and is convenient to operate.

The invention belongs to the field of traditional Chinese medicine, and discloses the application of aloe leaf powder in the preparation of a drug for treating chronic atrophic gastritis. Aloe leaf powder can significantly improve the phenotype of reduced number of stomach parietal cells, glandular dilatation, increased neck mucous cells and reduced chief cells in mice induced by tamoxifen, and has the efficacy of treating chronic atrophic gastritis. The invention discloses the application of at least one of aloe polysaccharide, aloe polyphenol or aloe flavonoid in the preparation of a drug for treating chronic atrophic gastritis. The polysaccharide, flavonoid and polyphenol in the aloe leaf powder all have the efficacy of treating chronic atrophic gastritis, and the ratio of polysaccharide>polyphenol>flavonoid. The invention discloses a composition for treating chronic atrophic gastritis, which is a composition of polysaccharide, polyphenol and flavonoid according to the mass ratio of (11.247-65.32):(0.373-4.551):(0.452-7.182), which can significantly improve the efficacy of chronic atrophic gastritis and has a synergistic effect.

The present invention provides a method for culturing an adherent cell, including a step of suspension culturing the adherent cell in a medium containing a nanofiber composed of a water-insoluble polysaccharide, wherein the culture is performed along with stirring.

The invention discloses a cell solution mixing structure which comprises a feeding cavity and a test tube, the feeding cavity comprises a sample injection needle through hole, a sample injection needle and a mounting hole, the sample injection needle through hole is formed in the feeding cavity in the vertical direction of the feeding cavity, the sample injection needle penetrates and is inserted into the sample injection needle through hole, the top of the sample injection needle is limited and fixed in the sample injection needle through hole through a sealing joint, and the test tube is arranged in the mounting hole. The mounting hole is formed in the feeding cavity in the transverse direction of the feeding cavity and is communicated with the sample injection needle through hole; the cell solution uniform mixing structure further comprises a vibration structure, the vibration structure is arranged in the mounting hole, and the vibration end of the vibration structure abuts against the sample injection needle. Wherein the vibration end of the vibration structure impacts the top end of the sample injection needle, and the bottom end of the sample injection needle amplifies vibration and then vibrates and uniformly mixes a cell solution in the test tube. The uniform mixing of the cell solution in the test tube is more friendly, and the problems of cell hanging on the wall, low cell survival rate and non-uniform distribution of cells in the solution caused by uniform mixing of the cell solution in the test tube through rotation of the motor in the prior art are solved.

The invention relates to a cell collection method for improving transmission electron microscope picture quality of cells cultured in vitro, and belongs to the field of transmission electron microscope biological sample preparation. Comprising the following steps: (1) reviving cells: discarding a drug-containing culture solution, and adding a fresh complete culture solution; (2) scraping cells and adding glutaraldehyde stationary liquid for pre-immobilization; (3) pre-fixation: sucking the supernate by using a needle cylinder, and adding 1mL of stationary liquid along the tube wall; (4) rinsing: rinsing for 5 minutes by using 0.1 M PB and pure water; (5) carbonization: putting the cell agglomerates into a carbonization cabin, and fumigating and burning the cell agglomerates for 1-1.5 hours by using 1% osmium tetroxide; (6) pure acetone induction: putting the sample obtained in the step (5) into 100% acetone for 5 minutes twice; (7) permeating: adding a mixed solution of epoxy resin and 100% acetone, centrifuging, and putting into a 38 DEG C oven for 2 hours; and (8) embedding and curing. According to the method, the cells can be collected in a short time, the influence of operation conditions and environment on the collection process is reduced, and the TEM picture quality of the adherent cells cultured in vitro is improved.

The invention relates to the technical field of surface plasma resonance, and discloses an establishment method and application of an intestinal wall cell mucous layer model. The method comprises the following steps: carrying out bionic treatment on the surface of a metasurface plasmon resonance chip, inputting a culture solution containing intestinal wall cells into the surface of the metasurface plasmon resonance chip through a micro-fluidic chip to carry out intestinal wall cell culture, inputting an intestinal tract simulation solution into the surface of the metasurface plasmon resonance chip, and carrying out intestinal tract simulation on the surface of the metasurface plasmon resonance chip. And detecting the refractive index change of the surface of the metasurface plasmon resonance chip, and monitoring the forming process of the mucus layer of the intestinal wall cell in real time according to the refractive index change. According to the intestinal wall cell mucous layer model obtained through the construction method, unmarked real-time continuous monitoring of the intestinal wall cell mucous layer forming process, dynamic observation of physiological changes of cells and simulation of the real intestinal environment can be achieved, and the method has high sensitivity and accuracy, is low in detection cost and has remarkable market prospects.

The present disclosure provides gene therapy vectors, such as recombinant adeno-associated virus (rAAV), produced in mammalian adherent cells cultured in suspension conditions, for expressing a human micro-dystrophin gene. The present disclosure also provides compositions and methods of using these rAAV to treat muscular dystrophy, such as, e.g., Duchenne Muscular Dystrophy.

The invention belongs to the technical field of cell clone acquisition, and discloses a needle head, a teasing needle and an adherent cell clone acquisition method.The needle head comprises a needle head body and a supporting column, the needle head body is provided with a suction channel, and the suction channel penetrates through the first end face, in the height direction, of the needle head body; the multiple supporting columns are arranged on the outer surface of the needle head body at intervals in the circumferential direction of the needle head body, and the supporting columns protrude out of the first end face; the adherent cell clone obtaining method comprises the steps that when cell clone obtaining is conducted, the needle body is placed in a culture solution of a culture container, all the supporting columns abut against the inner wall of the culture container, a flow field is more stable, then cell clone is sucked through the suction channel, and therefore when the cell clone is sucked, the cell clone is separated from the culture solution, and the adherent cell clone is obtained. The distance between the first end face and cell clone does not need to be noticed; meanwhile, the needle head can also form a relatively stable flow field on the surface, so that the acquisition efficiency of adherent cell clone is improved.

The invention relates to the field of biomedical engineering, and discloses a construction method and application of an intestinal wall cell barrier model. The method comprises the following steps: performing intestinal wall cell culture on a metasurface plasmon resonance chip of which the surface is subjected to bionic treatment in a culture medium to obtain a chip plate attached to the wall of intestinal wall cells, and monitoring the forming process of a barrier layer of the intestinal wall cells in real time by detecting the refractive index change of the surface of the metasurface plasmon resonance chip. The intestinal wall cell barrier model obtained by the construction method can realize unmarked real-time continuous monitoring of the formation process of the intestinal wall cell barrier layer and dynamic observation of physiological changes of cells, and the method has high sensitivity and stability.

The invention relates to a serum-free culture medium for suspension culture of McCoy cells and application of the serum-free culture medium. The serum-free culture medium for suspension culture of the McCoy cells comprises amino acid, vitamins, inorganic salt, glucose, sodium hyaluronate, PF68 and water. Wherein the serum-free culture medium meets at least one of the following characteristics (t1) and (t2): (t1) amino acid contains one or more of asparagine, cysteine, glutamic acid and proline; and (t2) the inorganic salt contains anhydrous disodium chlorine phosphate. The serum-free culture medium provided by the invention can realize high-density suspension culture of McCoy adherent cells without the help of any adsorption carrier, and can realize continuous and stable passage.

The utility model discloses a bioreactor microcarrier adherent cell pancreatin digestion device, which belongs to the technical field of cell pancreatin digestion, and comprises a pancreatin digestion unit connected with a bioreactor, and the pancreatin digestion unit is connected with a bottom valve of the bioreactor through a second connecting pipe; the micro-carrier adherent cells are digested by pancreatin in the bioreactor. The bioreactor microcarrier adherent cell pancreatin digestion device provided by the utility model can be used for observing the cell digestion state in real time and terminating digestion in time, carrying out data statistics and analysis on cell counting, calculating the recovery rate of digested cells, and observing the cell attachment rate and the cell growth condition of the cells in a next-stage bioreactor, so that the digestion efficiency of the cells in the next-stage bioreactor is improved. Compared with culture parameter data of a production batch for cell digestion by using a butterfly-shaped pancreatin digester before improvement, the recovery rate and the adherence rate are stably increased, and the working efficiency and the product yield are better improved.