234 results about "Microcarrier" 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 preparation method of a mesenchymal stem cell exosome based on microcarrier three-dimensional culture and application of the mesenchymal stem cell exosome in anti-aging skin care products. According to the method, a three-dimensional culture system of mesenchymal stem cells is optimized, and the anti-aging function activity of the subsequently separated and purified exosome is remarkably improved by adopting a microcarrier with recombinant collagen as a main component; subsequently, by optimizing an exosome separation and purification technology, and through a specific separation and purification mode and conditions, the exosome with anti-aging active ingredients is further obtained. Experiments prove that the exosome obtained by the method disclosed by the invention can effectively remove metalloproteinase 1 and promote collagen regeneration, has a remarkable anti-aging effect, is suitable for essence, face cream and other dosage forms, and has a wide market prospect.

The invention discloses a natural product-induced artificial nano-vesicle derived from human umbilical cord mesenchymal stem cells and application of the natural product-induced artificial nano-vesicle in ischemic diseases. The preparation method of the nano vesicle derived from the human umbilical cord mesenchymal stem cells comprises the following steps: S1, inoculating the human umbilical cord mesenchymal stem cells to a 3D microcarrier; then adding traditional Chinese medicine active ingredients to activate the human umbilical cord mesenchymal stem cells, continuing to culture, cracking the microcarrier, and harvesting cells; s2, dispersing the human umbilical cord mesenchymal stem cells obtained in S1 to obtain a suspension, extruding the suspension through a polycarbonate film, and then discarding large particles by adopting a gradient centrifugation method to obtain the nano-vesicles. Compared with naturally secreted exosomes, the exosome has the advantages that the yield is increased, the treatment efficiency is improved, the production cost is reduced, and compared with stem cells, the problems of low transplantation rate and the like are avoided. The stem cells are pretreated based on the traditional Chinese medicine, the vesicles are extracted by utilizing an extrusion method, and the yield and the treatment efficiency of the nano vesicles are remarkably improved.

The invention provides an elastic microcarrier as well as a preparation method and application thereof, and belongs to the field of water, wastewater and sewage treatment, the elastic microcarrier comprises the following components in parts by mass: 50-75 parts of thermoplastic elastomer, 25-10 parts of diatomite, 10-5 parts of tourmaline and 15-10 parts of iron-carbon powder. The material has good elasticity, adsorbability, biocompatibility and electrochemical characteristics, and can form a microscopic primary battery effect. The elastic microcarrier is applied to sewage treatment with a membrane bioreactor, pollutants can be removed through physical adsorption, chemical action and biological enhancement, meanwhile, the surface of a membrane is kept clean in combination with flexible scraping, membrane pollution is remarkably slowed down, the running membrane flux is improved, and the service life of the membrane is prolonged. The mechanical damage of the traditional rigid filler to the membrane is avoided, and the service life of the membrane is prolonged. The preparation method of the elastic microcarrier is simple and easy to operate, the elastic microcarriers with uniform specifications can be prepared in batches, and the applicability is high.

A preparation method for a micro-vehicle formed by enveloping siRNA-loaded polypeptide dendrimer nanogel with a cell membrane of engineered MSCs, and the use of same. The method comprises: using lentivirus-mediated gene transfection technology for constructing MSCs that highly express CXCR4 and PSGL-1; using cytokinin B for stimulating a cell to generate vesicles, and then performing centrifugation to collect the vesicles; by means of the electrostatic attraction between polypeptide dendrimer nanoparticles and siRNAs, attaching the siRNAs of IL-6 and HIF-1α into the polypeptide dendrimer nanoparticles; and by means of using a membrane extruder, enveloping the siRNA-loaded polypeptide dendrimer nanogel with the cell membrane. The prepared micro-vehicle exhibits a good property of aggregating at inflammatory blood vessel sites and a targeting function for joint cavities with rheumatoid arthritis. In addition, the loaded siRNAs can inhibit the expressions of IL-6 and HIF-1α in the joint cavities and inhibit the cell migration and angiogenesis. The micro-vehicle targeting inflammation sites has the advantages of excellent biocompatibility, ideal siRNA loading ratio, good inflammation targeting function, etc.

The present disclosure provides a porous microcarrier loaded with laccase, a preparation method and an application. The porous microcarrier loaded with laccase includes: a hydrogel porous microcarrier; laccase, which is loaded on the hydrogel porous microcarrier by covalent bonding; wherein the hydrogel porous microcarrier is a methacrylated hydrogel microcarrier filled with chitosan, providing a loading site for laccase.

The invention discloses a gelatin microcarrier lysate as well as a preparation method and application thereof. The gelatin microcarrier lysis buffer comprises lyase, a cell protective agent, a cell energy metabolism protective agent, an antioxidant, a buffer solution and a chelating agent. The gelatin microcarrier lysate has extremely strong protectiveness to cells, abandons damaging trypsin and EDTA, and innovatively adds a cell protection combination of a cell protective agent L-arginine, a cell energy metabolism protective agent sodium pyruvate and an antioxidant vitamin C, so that the cell survival rate (gt; 98%) and the retention rate (gt; the method is high in cell yield, good in safety and wide in range.

The invention belongs to the technical field of rabies vaccines, and particularly relates to a purification method of a rabies virus inactivated vaccine, which comprises the following steps: step 1, carrying out passage by using a Walvax-2 cell strain through a cell factory method, and carrying out multiplication culture on the passage production cells in a microcarrier bioreactor; 2, cleaning the cells with a treating fluid, inoculating the cells with a rabies virus fixed strain, and preparing and culturing a rabies virus solution; step 3, carrying out ultrafiltration concentration on the rabies virus stock solution; step 4, purifying the rabies virus concentrated solution; step 5, inactivating the rabies virus; and 6, hydrolyzing to obtain the rabies vaccine stock solution. By adopting the composite Capto Core 700 column chromatography, impure proteins can be effectively removed, the recovery rate is high, the quality of the rabies vaccine obtained by adopting a mode of firstly purifying and then adopting beta-propiolactone inactivation is excellent, and the effect of the finally obtained human diploid cell rabies vaccine stock solution is good.

The embodiment of the invention discloses a method for constructing a cartilage micro-tissue by compounding an extracellular matrix with a vascular matrix component and application. The method comprises the following steps: preparing a porcine decellularized cartilage extracellular matrix; generating a cartilage extracellular matrix suspension; carrying out screening treatment on the cartilage extracellular matrix suspension; preparing a porcine acellular cartilage extracellular matrix microcarrier; preparing a target suspension of a common system from the porcine acellular cartilage extracellular matrix microcarrier and vascular matrix component cells, and transferring the target suspension to a bionic floating microenvironment of a shaker bioreactor for dynamic culture; carrying out culture treatment on the porcine acellular cartilage extracellular matrix microcarrier; and taking out the cell-loaded porcine decellularized cartilage extracellular matrix microcarrier from the cultured and treated porcine decellularized cartilage extracellular matrix microcarrier as the constructed cartilage micro-tissue. According to the embodiment, the condition of secondary injury to a user can be improved and reduced, the repairing effect is improved, and the adaptability of the constructed cartilage micro-tissue is improved.

The invention discloses a stem cell bionic microcarrier amplification system to solve the problems of low efficiency, easy function loss and the like of a traditional amplification method. The core of the micro-carrier comprises a porous degradable polymer micro-carrier of which the surface simulates an extracellular matrix (ECM) topological structure and is covalently bound with ECM protein; in a customized bioreactor, a physiological range fluid shear force is generated by periodically regulating and controlling stirring, and an in-vivo mechanical microenvironment is simulated. According to the system, through the synergistic effect of an ECM bionic structure and dynamic mechanical stimulation, adhesion and proliferation of stem cells are effectively promoted, and stemness and multidirectional differentiation potential are maintained. The amplification multiple can reach 10 times or more of that of a traditional method within 7-10 days, the method has the advantages of being capable of being amplified, standardized, low in cost and the like, and a key technical support is provided for industrialization of stem cell treatment.

Provided are methods of obtaining large scale cultures of muscle-derived muscle precursor cells (MPCs) using microcarriers as growth matrices, methods for obtaining therapeutically effective amounts of these cells, populations of cells obtained by the methods, and compositions comprising expanded cells, and a method for preparing a medicament, for example for the treatment of skeletal muscle dysfunction.

The invention relates to image differentiated multiplex assays. Provided herein are encoded microcarriers for analyte detection in multiplex assays. The microcarrier is encoded using an analog code for identification and includes a capture agent for analyte detection. Also provided are methods of making the encoded microcarriers disclosed herein. Further provided are methods and kits related to multiplex assays using the microcarriers described herein.

The invention relates to a polylactic acid microcarrier and a method for cell amplification by using the polylactic acid microcarrier. The preparation method disclosed by the invention is simple and convenient to operate, the prepared polylactic acid microcarrier is uniform in particle size and pore size distribution, the microcarrier is subjected to surface retreatment, more active sites are provided for cell adhesion, a three-dimensional matrix environment closer to a body is provided, cell amplification is facilitated, the dryness and differentiation potential of stem cells are maintained, and the preparation method is suitable for large-scale popularization and application. And the microcarrier can be repeatedly used, so that the use cost is reduced.

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

Devices for filtering liquids are provided herein. More particularly, the devices may be devices for separating biological cells from microcarriers the cells have been grown with, as well as other debris and materials, that are found in the cell culturing liquid. The devices may also be foldable or collapsible.

The invention discloses a preparation method of extracellular small vesicles of functional homogeneity mesenchymal stem cells. The preparation method comprises the following steps: separately and statically culturing tissues from different individual sources for 8-12 days through a 3D culture microcarrier; and carrying out mixed dynamic culture on the culture product obtained in the step S100, collecting a supernatant, and separating the extracellular vesicles from the supernatant. The sEVs obtained by the preparation method have high-homogeneity mesenchymal stem cell sEVs proteome, and in addition, compared with sEVs purified by independently culturing mesenchymal stem cells, the sEVs have stronger immune regulation and control capabilities (including inflammatory reaction inhibition, proinflammatory factor reduction and inflammatory inhibition factor increase) and regeneration and repair promotion capabilities.

Disclosed in the present invention are a 3D preparation method for and a use of hepatocytes. In the present invention, hepatic precursor cells are differentiated into functional hepatocytes via low-oxygen conditions and a specific culture medium, and after sampling and counting, hepatic spheroids are subjected to liquid nitrogen cryopreservation. The 3D preparation of hepatocytes does not require a microcarrier, and digestive enzyme digestion is not needed.

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.

The present invention relates to a single-phase, spherical edible microcarrier for the three-dimensional culture of stem cells intended for the preparation of cultured meat. The microcarrier comprises a homogeneous mixture of edible ionic polysaccharide (0.375-1.5%), edible protein (1-2.5%), edible polymer (0.025-0.1%) and water (95.9-98.6%). The invention also relates to a method for preparing these microcarriers by mixing three solutions: ionic polysaccharide in saline solution, edible protein in saline solution and edible polymer in water. The mixture is subjected to a process involving microspraying or electrospraying with calcium chloride in order to form spherical microcarriers of 50-300 micrometres. The design of the single-phase, spherical edible microcarrier simplifies the production of cultured meat by dispensing with the need for core-shell structures, thereby improving cell anchorage efficiency and facilitating cell recovery.

Provided are a dental pulp mesenchymal stem cell-microcarrier scaffold complex, and a preparation method therefor and a use thereof. The dental pulp mesenchymal stem cells and a microcarrier are subjected to 3D culture and amplification under low oxygen and high osmotic pressure conditions, and then sampling and counting are carried out, and a dental pulp mesenchymal stem cell-microcarrier scaffold complex is directly subjected to stem cell liquid nitrogen cryopreservation. The dental pulp mesenchymal stem cell-microcarrier scaffold complex can secrete higher cytokines and exert better anti-inflammatory and immunomodulatory effects. The complex can be directly cryopreserved without degradation of a scaffold and preparation of single cells, so that the operation is simple, and the activity and the survival of the cells which are resuscitated from cryopreservation are higher. Before cell treatment, the dental pulp mesenchymal stem cell-microcarrier scaffold complex is directly thawed, has auxiliary materials added, and is placed into a biological artificial liver reactor for treating acute liver failure.

A system, method, and apparatus for delivering genetic material to a shoot apical meristem. The delivery system may have a biolistic particle delivery assembly having a gas acceleration tube and a bombardment chamber. The bombardment chamber has a microcarrier launch assembly comprising an opening adapted to receive gas for acceleration of the microcarrier system and to direct gas to a target. The delivery system may have a source of pressurized carrier gas operatively connected to a first end of the gas acceleration tube; and an extender. The extender has a channel configured to fit through the opening of the microcarrier launch assembly. The length of the extender facilitates the penetration of the microcarrier system to an L2 layer of the shoot apical meristem, conferring germline transformation and / or modification.

To provide a microcarrier that enables confirmation of contamination.SOLUTION: The problem is solved by a microcarrier having an acid-base indicator fixed on its surface. It is more preferable that an amount of the acid-base indicator fixed on the microcarrier is 0.1 μg / cm2 to 900 μg / cm2.SELECTED DRAWING: None

A three-dimensional culture method for large-scale preparation of stem cells, comprising a three-dimensional microcarrier-based cell resuscitation method, a three-dimensional microcarrier cell culture-based in situ passage method, a three-dimensional microcarrier in situ freeze preservation method for cells, a three-dimensional microcarrier cell adsorption culture method, a method for harvesting cells on a three-dimensional microcarrier, a method for sampling cells cultured on a microcarrier, and a three-dimensional microcarrier-based cell large-scale expansion method.

This invention belongs to the field of biomedical engineering technology, specifically the field of cell function detection technology, and relates to a method for evaluating the ability of mesenchymal stem cells (MSCs) to promote Treg cell proliferation. This method utilizes porous microcarriers to achieve three-dimensional cell distribution, employing low-speed rotation throughout the culture process to simulate the in vivo suspension microenvironment. A culture medium matching human physiological parameters is configured, with oxygen concentration adjusted to a hypoxic range of 1%-3% to simulate the microenvironment of inflammatory sites. Mononuclear cells with a purity of over 95% are obtained through magnetic bead sorting and co-cultured with MSCs for approximately 3 days, significantly shortening the culture period and reducing data deviation, thus achieving specific screening of MSCs with high immunomodulatory functions. By constructing a co-culture microenvironment that conforms to human physiological characteristics, this invention effectively improves detection sensitivity and repeatability, avoids interference from individual differences in peripheral blood mononuclear cells from different sources, and provides technical support for the quality control and clinical application of MSCs.

The invention discloses a kelp gametophyte gene editing method and application, and belongs to the technical field of gene editing, the kelp gametophyte gene editing method comprises the following steps: preparing a kelp gametophyte receptor; assembling an RNP complex and preparing a microcarrier; introducing the RNP compound into a kelp gametophyte by using a gene gun; and recovering culture and screening. According to the technical scheme, the dependency of a microinjection technology on precise instruments and professional operation skills is eliminated, efficient gene delivery can be achieved through a standardized gene gun operation process, the technical threshold is greatly reduced, and the kelp gametophyte gene editing method has remarkable technical advantages. According to the method, the high-throughput characteristic of a gene gun technology is fully exerted, multiple mutants can be obtained through one-time bombardment, the experiment efficiency is greatly improved, and powerful support is provided for kelp large-scale gene function research and molecular breeding.

The application discloses a kind of double-network amphiphilic hydrogel microcarriers capable of maintaining cell stemness and application thereof, and belongs to the technical field of cell culture.The present application copolymerizes SBMA with acrylamide (AAm) monomer to form a double-network structure with acrylic acid (PAA) to prepare hydrogel microcarriers.The double-network structure hydrogel microcarriers PSAm obtained based on amphiphilic materials can effectively expand MSCs while maintaining their cell stemness for a long time, which is conducive to long-term culture of MSCs.Compared with ordinary culture, the artificial subculture process can be reduced, manpower and material resources can be saved, resource waste can be avoided, and a new experience method is provided for long-term culture, preservation and maintenance of cell stemness of MSCs.

The invention discloses a method for large-scale cryopreservation of animal cells. The method comprises the following steps: carrying out first-stage amplification on to-be-preserved cells under the support of matrigel; performing second-stage amplification on the to-be-preserved cells subjected to the first-stage amplification under the support of a microcarrier; mixing the to-be-preserved cells with a serum-free cryopreservation solution to prepare a micro-capsule suspension; and putting the microcapsule suspension into a cryopreservation container to carry out cryopreservation. According to the invention, through amplification of two stages, the to-be-preserved cells have reached a high density of 109 orders of magnitude before cryopreservation, a re-amplification link in a cell recovery stage can be greatly shortened, and the to-be-preserved cells are subjected to microencapsulation through the micro-fluidic chip and then cryopreserved, so that physical protection is given to the to-be-preserved cells; the damage of ice crystals in the cryopreservation process is reduced, the survival rate after resuscitation is improved, and the microencapsulated cells can also be directly subcultured, so that the resuscitation operation is simplified; according to the cell cryopreservation device, the cryopreservation container with a large volume is used, so that the cost of cell cryopreservation is reduced.

A substrate for culturing cells that comprises a modified prolamin having a net positive charge is provided. The modified prolamin may be a prolamin that has been modified through amidation or esterification, such as the addition of a methyl-ester group or an ethanolamine group. The modified prolamin is animal-free may be food-grade and / or may be free of cross-linkers. The modified prolamin may be in a coating that can be applied to a surface for culturing adherent cells, such as a microcarrier (e.g., beads) or well plates. Substrates coated with the modified prolamin are able to support cell expansion and can be used for food-grade applications such as cultured meat.

The invention provides a composite conditioner for acid soil remediation and application thereof, and belongs to the technical field of agricultural environment and soil remediation. The composite conditioner comprises a first component, a second component and a third component which are independently packaged, wherein the modified straw biochar in the first component is used as a'microcarrier 'and a'buffer', calcium magnesium ore powder containing high-activity calcium magnesium oxide is loaded, slow release of alkaline substances is achieved, local over-alkalinity and hardening risks are avoided, and the porosity and the fertilizer retention capacity are improved; the second component is compounded with high-molecular polymer hydrolytic polymaleic anhydride, polyaspartic acid and polyglutamic acid according to a specific proportion, and the synergistic chelation capacity to aluminum and manganese ions and the dispersion capacity to soil particles are obviously superior to those of any single component and other proportion combinations; the third component selects an acid-resistant compound strain and is supplemented with amino acid to serve as a nutrient substance for rapid proliferation of the acid-resistant compound strain, so that high survival rate, colonization success rate and functional expression efficiency of the exogenous flora in an acidic stress are ensured.