665results about "General culture methods" patented technology

The present disclosure provides a Chinese Hamster Ovary (CHO) cell comprising one or more exogenous nucleotide sequences, wherein the one or more exogenous nucleotide sequences is inserted into a chromosomal locus of the CHO cell, wherein the chromosomal locus is located within contig NW_023276805.1 of chromosome X. Also provided herein are methods of producing a CHO cell capable of expressing a sequence of interest, methods of expressing a sequence of interest comprising culturing the CHO cell as described herein, and kits comprising the CHO cell as described herein.

The invention discloses a preparation method of DNA (Deoxyribonucleic Acid) hydrogel with programmable mechanical strength. By regulating and controlling the molar ratio of the catalytic initiating chains to the total amount of the DNA monomer building blocks, multiple hydrogels which are equivalent in mass concentration and have different mechanical strengths are constructed. According to the method, a strategy similar to catalytic assembly is adopted, the feeding molar ratio of a catalytic initiating chain to a DNA monomer building block is creatively adjusted, the proportion of cross-linking reactions at different levels is regulated and controlled, then the DNA hydrogel with different mechanical strengths is synthesized under the isothermal condition, and the storage modulus range is about 200-2000 Pa. In addition, the change of the mechanical strength is independent of the total concentration of the matrix (DNA) of the hydrogel, the DNA sequence, the stress relaxation rate and other properties. The method provides a novel convenient method for constructing DNA hydrogels with different mechanical strengths, and possibly provides a novel tool for researching the specific influence of mechanical properties of materials on cell / tissue behaviors.

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ / Activin / Nodal signaling and BMP signaling are provided. Also provided are methoc and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

Disclosed are methods for preparing non-filamentous scaffolds (e.g., sheets) for cell or tissue culture. These methods can comprise providing at least a first printing composition (e.g., a bioink) and a second printing composition (e.g., a bioink or a fugitive ink); chaotic printing the first printing composition and the second printing composition to generate a microstructured precursor comprising a plurality of lamellar structures formed from the first printing composition and the second printing composition; extruding the microstructured precursor through a nozzle (e.g., a fan-shaped nozzle, a curved fan-shaped nozzle, or an annular nozzle) to produce a non-filamentous microstructured precursor; and curing the non-filamentous microstructured precursor to provide the non-filamentous scaffold for cell or tissue culture.

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

The present disclosure provides solid collagen constructs and tissue compositions, wherein a polymerizable collagen solution or suspension is extruded in the presence or absence of cells to formed an aligned architecture comprising solid collagen constructs such as those made with fibrillar collagen. Methods of using and of manufacturing solid collagen constructs and tissue compositions, where the component collagen is solid fibrillar collagen and cells are preferentially aligned, are also provided.

The present invention is concerned with a manufactured article prepared from a biomaterial comprising isolated and purified type I collagen fibrils derived from abalone and a method of preparing such an article. The manufactured article can be prepared by 3D bioprinting, and therefore the present invention also envisages a novel bioink for a 3D bioprinter comprising an isolated and purified type I collagen fibril and cartridges containing such a bioink. The invention additionally relates to a method of isolating type I collagen fibrils from abalone. The manufactured article is useful in a number of biomedical applications including as an implantable device where it can function as a 3D bioprinted hard or soft tissue scaffold for tissue engineering.

A method of modifying a surface of a three-dimensional (3D) article includes immersing at least one part of the 3D article in a buffered solution of functionalized peptides, allowing reaction between the functionalized peptides and reactive groups on the surface of the 3D article; and washing the at least one part of the 3D article to remove unreacted functionalized peptides. The surface-modified 3D article includes a plurality of peptides covalently bonded to the surface of the 3D article via a cysteine bridge. The surface- modified 3D article can be used as a scaffold for the formation of biological tissue or bodily implants.

The present invention provides constructs including a tubular biodegradable polyglycolic acid scaffold, wherein the scaffold may be coated with extracellular matrix proteins and substantially acellular. The constructs can be utilized as an arteriovenous graft, a coronary graft, a peripheral artery bypass conduit, or a urinary conduit. The present invention also provides methods of producing such constructs.

A method for in situ differentiation of stem cells within a cell culture bioreactor is provided. The method includes providing a bioreactor vessel having a cell culture chamber, the cell culture chamber having an inlet for introducing fluid into the cell culture chamber and an outlet for introducing fluid out of the cell culture chamber. The bioreactor vessel also includes a cell substrate disposed within the cell culture chamber for culturing cells on the cell substrate. The method includes seeding undifferentiated stem cells onto the cell substrate within the cell culture chamber and perfusing the cell culture chamber with a differentiation medium to promote differentiation of the undifferentiated stem cells into specific cell lineages, thereby causing the undifferentiated stem cells to become differentiated cells.

The cell-containing vessel includes a cell culture vessel, a cell culture medium contained in the cell culture vessel, a first cell layer formed on a cell culture surface of the cell culture vessel, and a second cell layer that contains stroma-forming cells and is formed on the first cell layer. The second cell layer is removable from the first cell layer.

The present disclosure relates to a transplantable cell therapy product composition for diabetes mellitus that contains an aggregate of insulin-secreting cells derived from stem cells. In the present disclosure, an NF microwell array membrane was fabricated by applying a molding process to an electrospun, permeable, biodegradable polycaprolactone (PCL) NF membrane and thus allows gases and soluble factors to permeate therethrough. The NF microwell of the present disclosure could provide more nutrients to the iPSC aggregates than conventional impermeable PDMS microwells, thus enhancing survival and differentiation capabilities of the cells. Additionally, the NF membrane was attached singly to the subcutaneous tissue and to the surface of organs such as liver and peritoneum without the need for a fixing material or separate sutures and was integrated with surrounding tissues, resulting in higher insulin secretion than PDMS microwells. Therefore, the present disclosure can be effectively utilized as a composition for the prevention or treatment of diabetes.

A cell culture microcarrier bead is proposed. The microcarrier bead comprises a bead body having its surface flecked with plasmonic nanoparticles. In a second aspect, the invention relates to a cell culture reactor, containing a cell culture medium and the proposed microcarrier beads. A third aspect of the invention concerns a method for observing living cells on such microcarrier beads. Yet a further aspect of the invention relates to a method for packing nanoparticles on a carrier body.

The invention provides a method for preparing an ordered cell-containing microarray, and a system for preparing an ordered cell-containing microarray.

The application pertains to compositions and methods useful for growing living cells such as stem cells. The compositions employ a mixture of an extracellular matrix and discrete carbon nanotubes. The extracellular matrix may also comprise components selected from the class of proteins, proteoglycans, polysaccharides, lipids, peptides, messenger molecules, signaling molecules, or any mixture thereof. The discrete carbon nanotubes are usually less than about 1% by weight of the dry weight of the total composition.

The invention provides porous silk fibroin microspheres as well as a preparation method and application thereof, and belongs to the technical field of biomedical materials. According to the method, a high-molecular compound containing a hydrophilic functional group is selected as a specific inducer, a silk fibroin solution and the specific inducer are blended, the intermolecular interaction of silk fibroin is regulated and controlled under the action of the inducer, and a large number of silk fibroin microspheres which are uniform in size and stable in structure are formed in a self-assembly mode in the concentration-film forming process. On the basis, the inducer is removed, and then drying treatment is performed to form the porous silk fibroin microspheres with high purity and high specific surface area. The preparation process is simple, no toxic reagent is used in the preparation process, and the obtained silk fibroin microspheres can be applied to the fields of cell culture carriers, drug delivery systems, protein separation and purification and the like and have wide application prospects in the field of biomedical materials.

According to the present invention, the present invention relates to a polymer microparticle having a core-shell structure, which comprises: a core comprising a first biocompatible polymer, a metal ion, and an organic crosslinker having one or more reactive functional groups; and a shell surrounding all or a part of the core and comprising a second biocompatible polymer, a metal ion, and an organic crosslinker having one or more reactive functional groups; and a cell attachment-inducing layer formed on the surface of the polymer microparticle; and a microcarrier, a cell complex comprising the same, a medical composition, a cosmetic composition, a medical product, and a cosmetic product.

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.

A microplate including a plurality of wells for receiving a dome of a supporting matrix, each well of the plurality of wells comprising a bottom surface having at least one retention structure to provide mechanical retention of one of the assay samples, the at least one retention structure comprising a diameter ranging between about 0.5 millimeters and about 5 millimeters and a height, and wherein a ratio of the height to the diameter ranges from about 10% to about 30% and a method of using the same.

To provide a cell aggregate producing substrate, a method for manufacturing same, a method for producing a cell aggregate, and a method for improving cell utilization efficiency when producing a cell aggregate.　Provided is: a cell aggregate production substrate provided with, on a substrate capable of suppressing adhesion of cells, a plurality of spots each formed of a polymer including a repeating unit derived from a monomer represented by formula (I) [in the formula, Ua1, Ua2, Ra1, and Ra2 are each as described in the description and claims]. The proportion of the total area of the spots with respect to the surface area of the substrate is 30% or more. The diameter of the spots is 50-5000 µm, and the interval between the spots is 30-1000 µm. Also provided are a method for manufacturing the cell aggregate production substrate, a method for producing a cell aggregate using said substrate, and a method for improving cell utilization efficiency when producing a cell aggregate.

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.

Provided herein are compositions comprising a particle comprising a library encoded bead (e.g., DEL bead) coated in a polymeric matrix forming a core-shell particle, wherein the core-shell particle is further inside a 3D tissue culture. Also provided herein are methods of making such particles, and methods of using such particles for drug discovery such as identification of bioactive compounds via high-throughput cellular activity-based phenotypic screens of DNA-encoded chemical library beads.

The invention relates to hydrogels comprising the crosslinked dextran polymers of the invention and / or hydrogels further comprising biological cells. The invention also relates to therapeutic uses of the hydrogels of the invention for treating disorders or diseases in mammals, the disorders or diseases resulting from lack of or insufficient endocrine function of the pancreatic organ, and to implants comprising the hydrogels of the invention. The invention also relates to implants comprising the hydrogels of the invention and further comprising rings and nets.

The present invention relates to a method for cultivating mammalian cells comprising the step of incubating mammalian cells in a culture medium comprising at least one sodium salt, at least one potassium salt and at least calcium salt under an atmosphere comprising less than 1% carbon dioxide.

The present invention relates to novel cell culture and transfection methods using a wildtype Caf1 ("Caf1-WT") polymer or a variant thereof. Use of a Caf1-WT polymer or a variant thereof to increase cell viability, cell proliferation, cell migration and / or cell clustering; promote the formation of a 3D cell structure; and / or promote suspension cell culture of cells that are normally adherent are also provided herein.

The present invention addresses the problem of providing: a cell culture scaffold from which a cell culture can be recovered without being destroyed after culturing cells; and a method for producing a cell culture.　The cell culture scaffold according to the present invention is provided with a base having a water-repellent surface and a cell adhesion molecule formed on the water-repellent surface of the base. In addition, the method for producing a cell culture according to the present invention includes a step for using the cell culture scaffold to culture cells on the surface of the cell adhesion molecule. The cell culture scaffold according to the present invention can allow cells to be recovered in a spheroid shape without being damaged.

The present disclosure relates to a method for producing leather comprising the step of culturing cells on honeycomb macroporous polymeric scaffold. The present disclosure also relates to leather obtainable by this method.

A cell culture substrate according to the present disclosure is a cell culture substrate including at least one well including a side surface and a bottom surface, in which the side surface includes a first side portion defining a first space and a second side portion defining a second space communicating with the first space, and in plan view, an upper end portion of the first side portion is formed in a C shape having a central angle of 240 degrees or more, the second space extends from an opening portion of the upper end portion in a first direction away from the first space in the plan view, and the second side portion includes a facing surface facing the opening portion in the first direction, the facing surface being disposed at a position where a maximum distance between the opening portion and the facing surface in the first direction is smaller than a diameter of the C shape of the upper end portion in plan view.

The present disclose relates to a high-reactivity hydrogel particle and preparation method therefor and use thereof. The preparation method comprises the following steps: (1) performing polymerization reaction of double-bond modified polyethylene glycol, a modified natural polymer and an optional active functional monomer to obtain a gel; (2) smashing the gel to obtain a gel particle; and (3) performing enzymolysis reaction of the gel particle and an enzyme to obtain the hydrogel particle with reactivity. The double-bond modified polyethylene glycol has a double bond and a polyethylene glycol chain segment, the modified natural polymer has sulfydryl or a double bond, the active functional monomer is selected from a sulfydryl functionalized biological adhesive peptide or sulfydryl functionalized cholesterol, and the enzyme is an enzyme corresponding to the natural polymer. The obtained hydrogel particle has high reactivity and can be used for subsequent secondary crosslinking. The hydrogel particle also has good biocompatibility.

The present disclosure relates to a method for producing three-dimensional cell clusters on an inorganic cell culture platform comprising a three-dimensional structure, preferably a fractal structure. Such three-dimensional structures are preferably useful for culturing cells and tissues in three dimensions. Such three-dimensional structures are preferably useful for inducing differentiation of non-embryonic stem cells. In particular, such three-dimensional (3D) structures are useful for culturing primary tissue cells.