299 results about "Microcarrier" patented technology

The invention relates to a cell culture microcarrier as well as a preparation method and application thereof. The cell culture microcarrier comprises a polymer porous microsphere with an inside-outside through pore channel structure. The cell culture micro-carrier comprises a biodegradable micro-carrier and also comprises a non-degradable micro-carrier, so that the growth of cells in the carrier is facilitated, and the mass transfer of nutrient substances and metabolites is also facilitated, thereby promoting the growth of the cells, enabling the cultured cells to keep good activity and function, and being high in proliferation efficiency; the method is suitable for three-dimensional culture of somatic cells, stem cells, tumor cells and the like.

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 present invention belongs to the technical field of bioreactors, and in particular relates to a high-density culture method for porcine myoblasts based on a bioreactor. Through the steps of resuscitating porcine myoblasts, cell-microcarrier attachment treatment, bioreactor stirring mode conversion, counting and centrifugal cell collection, the porcine myoblasts are difficult to reach a high density in a bioreactor, and a harvest density of 10^6 cells / mL is achieved in the bioreactor, meeting the requirements of cell culture meat production on cell yield; at the same time, the culture time is shortened by more than 20%, greatly saving energy consumption cost and time cost.

A filter assembly for separating microcarriers from a fluid medium includes a collapsible container bounding a sterile compartment adapted to hold a fluid. An inlet port is attached to the container through which fluid flows into the compartment. An outlet port is attached to the container through which fluid flows out of the compartment. A filter is disposed within the compartment, the filter dividing the compartment into an inlet chamber that is fluidly coupled with the inlet port and an outlet chamber that is fluidly coupled with the outlet port, the filter allowing a medium to pass therethrough but preventing microcarriers disposed in the medium from passing therethrough.

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 relates to the field of biological medicine, in particular to a water-soluble alginate microcarrier as well as a preparation method and application thereof. The alginate microcarrier comprises a carrier core structure and a shell surface modifier, the micro-carrier core layer is formed by carrying out plasma crosslinking on water-soluble alginate, ethylenediamine tetraacetic acid salt, nitrilotriacetic acid salt, diethylene triamine pentaacetic acid and water-soluble divalent metal salt, and light-sensitive alginate and a photoinitiator can also be added, so that the micro-carrier core layer has two crosslinking networks; the shell layer is composed of one of gelatin, collagen, fibrinogen, albumin and fibrin, or the gelatin, the collagen, the fibrinogen, the albumin and the fibrin and other biocompatible polymers; the obtained microcarrier has the characteristics of high transparency and rapid degradation, and adherent cells can be amplified in vitro; the microcarrier obtained in the invention can be used for mesenchymal stem cell culture, and can be rapidly degraded under a mild cell condition to harvest cells.

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.

Edible microcarrier for growing anchorage-dependent cells to prepare a cultured meat product, where the edible microcarrier includes a microcarrier core and a microcarrier coating at least partially directly or indirectly covering the surface of said microcarrier core, wherein said microcarrier core is a hydrogel having only the following components: (A) 0.03-10.0 wt % of at least one ionically crosslinked polysaccharide; (B) 0.05-5 wt % of a mechanical stability agent different from (A); (C) 0.0-10 wt % of additives different from (A) and (B); (D) 0.0-50 wt % living cells; (E) 75-99.92 wt % of water; where the sum of (A)-(E) making up 100 wt %, and where the microcarrier coating includes a biopolymer, where the biopolymer is crosslinked using a crosslinking agent.

The invention discloses a novel oral delivery microcarrier with an intestinal adhesion function and a preparation method thereof, and the preparation method specifically comprises the following steps: preparing methacrylic anhydride gelatin, o-nitrobenzaldehyde group gelatin, a photoinitiator, a color indicator and a drug to be loaded into a mixed solution, and preparing microspheres by using a microfluidic instrument, and irradiating with an ultraviolet light source until curing to obtain the novel oral delivery microcarrier with the intestinal adhesion function. According to the invention, the limitation of the existing traditional single material is overcome, the compounding ratio of GelMA to GelNB is optimized, and the stability and biological activity of the microspheres are ensured; drug molecules are successfully and uniformly entrapped in the microspheres through an advanced microsphere preparation microfluidic technology; the targeting delivery efficiency of the medicine is improved, the healing and regeneration of tissues are promoted, and a new strategy and a new tool are provided for disease treatment and tissue engineering application.

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 invention discloses a porous microcarrier capable of loading human umbilical cord mesenchymal stem cells and a preparation method and application of the porous microcarrier, and relates to the technical field of biomedical engineering. Comprising the following raw materials in parts by weight: 0.1-5 parts of methylacryloylated gelatin, 0.1-5 parts of methylacryloylated silk fibroin and 0.1-5 parts of methylacryloylated chondroitin sulfate. The preparation method of the porous microcarrier comprises the following steps: S1, constructing a microfluidic platform; s2, preparing microsphere liquid drops; s3, preparing a porous microcarrier containing gelatin, silk fibroin and chondroitin sulfate; according to the prepared gelatin-silk fibroin-chondroitin sulfate microcarrier capable of loading the hUMSCs, the silk fibroin and the chondroitin sulfate are doped in the gelatin, and compared with the gelatin microcarrier, by adjusting the physical performance of the gelatin-silk fibroin-chondroitin sulfate microcarrier capable of loading the hUMSCs, the anti-inflammatory capacity and the immune regulation capacity of the hUMSCs can be effectively improved, and the capacity of stem cells for treating related diseases is improved.

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 belongs to the field of biological pharmacy, and discloses an MSCs (mesenchymal stem cells) culture process for improving the yield of protein polymers, which comprises the following steps: resuscitating and inoculating mesenchymal stem cells, transferring the mesenchymal stem cells into a culture container added with a microcarrier for rotating or shaking culture, and aseptically transferring the cultured mesenchymal stem cells into a culture bag; performing irradiation stimulation on the culture bag; after stimulation is completed, the mesenchymal stem cells are split, separated and purified, and the protein polymer is obtained. According to the MSCs culture process of some examples, more MSCs can be obtained, and more protein polymers can be obtained through purification.

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.

To provide a method for culturing stem cells with improved efficiency by using a gel that has improved structural strength and enables long-term and stable release of contents.SOLUTION: A bilayer composite hydrogel microcarrier includes: an inner layer gel structure obtained by ionically crosslinking an inner layer polymer with an inner layer crosslinker; and an outer layer gel structure obtained by covalently crosslinking an outer layer monomer with an outer layer crosslinker, which covers the inner layer gel structure; a first inner layer polymer being sodium alginate, a second inner layer polymer being carboxymethylcellulose, and a nutritional component being located inside the inner layer gel structure. The growth rate of stem cells is improved by controlling the composition of the bilayer composite hydrogel microcarrier, the weight ratio of sodium alginate to carboxymethylcellulose, and the location of the nutritional component. The structural strength can be ensured by controlling the weight percentage of carboxymethylcellulose.SELECTED DRAWING: Figure 2

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.

The invention discloses an in-vitro suspension culture medium and a preparation method thereof, and relates to the technical field of cell culture, the in-vitro suspension culture medium comprises amino acid, vitamin, inorganic salt, biogenic amine, nucleotide, fatty acid, lipoid, hormone, saccharide, protein, polypeptide and water, and the polypeptide comprises recombinant human collagen peptide. According to the invention, in-vitro large-scale cell suspension culture without a microcarrier is realized.

Provided herein is a method for preparing an emulsion, comprising the steps of allowing a dispersed phase liquid to flow from one side to the other side of a multi-hole plate through a plurality of micro-wells of the multi-hole plate while allowing a continuous phase liquid to flow, parallel to the multi-hole plate, on the other side of the multi-hole plate, shearing the dispersed phase liquid passing through the multi-hole plate to form liquid microspheres in the flowing continuous phase liquid. Further provided herein are an apparatus for preparing an emulsion and a process system for preparing microcarrier particles, which can be used for preparing emulsions and microcarrier particles on a large scale.

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.

The utility model discloses a full-automatic microcarrier cleaning device which comprises a cleaning tank, a liquid storage barrel, a waste liquid barrel, a collecting bottle, a magnetic stirrer, an electronic scale and a controller, the magnetic stirrer is arranged on the top of the cleaning tank, the cleaning tank is arranged on the upper portion of the electronic scale, and the lower portion of one side of the cleaning tank is connected with a silicone tube connector. The silica gel pipe connector is connected with a liquid inlet pipe A and a liquid outlet pipe A through liquid inlet and outlet pipes, the liquid inlet pipe A and the liquid outlet pipe A are correspondingly communicated with a liquid storage barrel and a collecting bottle respectively, the liquid storage barrel is further communicated with the upper portion of a cleaning tank through a liquid inlet pipe B, and the bottom of the cleaning tank is communicated with a waste liquid barrel through a liquid outlet pipe B. A filter screen is further arranged on the inner bottom wall of the cleaning tank. Corresponding electromagnetic valves are respectively arranged on the liquid inlet pipe A, the liquid outlet pipe A, the liquid inlet pipe B and the liquid outlet pipe B; and the magnetic stirrer, the electronic scale and the electromagnetic valves on the pipelines are electrically connected with the controller. The utility model has the advantages that the cleaning time is shortened, the operation is simple, the cleaning efficiency is improved, the loss of cell viability is reduced, and the consistency and stability of the process are kept.

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.