992results about "General culture methods" patented technology

There has been demand for an additional method for producing antibodies. The present invention provides: a polymer-coated crosslinked alginate gel fiber in which a core layer containing a crosslinked alginate gel and either antibody-producing cells (e.g., antibody-producing CHO cells) or bioactive-substance-producing cells (e.g., MIN6 cells derived from pancreatic β cells) is coated with a cationic polymer; and a method for producing antibodies, a bioactive substance, etc., using the fiber.

A device for modelling physiological and pathophysiological states of the brain is provided. The device comprises a hydrogel with micropattern channels having a length, width and depth. The micropattern channels comprise cells selected from the group consisting of neurons, astrocytes, microglia, oligodendrocytes, neuronal organoids, cancer cells, cancer spheroids, brain tumoral cells and combinations thereof.

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 of evaluating functional activity of an engineered three-dimensional tissue, methods of identifying a compound that modulates tissue function, and methods for identifying a compounds that is useful for treating or preventing a disease affecting tissue function are provided herein. Some methods include providing or obtaining a three-dimensional tissue scaffold defining a lumen or a cavity, the tissue scaffold comprising one or more polymeric fibers each having a micron-scale or nanometer-scale diameter, at least some of the one or more polymeric fibers encircling the lumen or cavity at a helical angle with respect to a longitudinal axis of the lumen or cavity, at an azimuthal orientation with respect to the longitudinal axis of the lumen or cavity, or at both.

The disclosed in vitro systems allow for multiple organs (organoids) to be cultured in the same conditions, connected with the same microcirculation, subjected to the same pathological state, and treated with the same therapeutic approach. This allows for determination of the how various organ systems respond to a pathological stress and also how a therapeutic approach may effect various organoids differently. The in vitro systems also provide for single organoids.

A method of culturing cells, the method including providing a microfluidic device including a substrate including an array of grooves that are substantially parallel; introducing cells into one or more of the grooves; providing a flow of culture medium over the groove array in a first flow direction that is across the grooves; culturing the cells under conditions to support viability, growth or differentiation, optionally wherein: (i) the cells are introduced into one or more grooves by flow over the groove array in the first flow direction; and / or (ii) the flow of culture medium is provided at a flow rate equivalent to 0.2-5 mL per million cells per day; and / or (iii) the cells are cultured for 1-14 days.

To provide a novel technique for detaching adherent cells from a solid phase.SOLUTION: A cell manipulation method includes: a step of culturing cells in a liquid; a step of disposing a flow path through which a gas is able to be introduced in the liquid; a step of forming an air bubble at an end portion of the flow path; and a step of attaching the cells to the air bubble.SELECTED DRAWING: Figure 10

The invention provides a hydrogel micropore array as well as a mold and application thereof, and belongs to the technical field of cell culture. The hydrogel micropore array and the die thereof provided by the invention can be used for three-dimensional formation and culture of cell microspheres and organoids with controllable quantity, position and size, and further can be used for high-quality high-content imaging. According to the hydrogel micropore array preparation mold provided by the invention, a plurality of hydrogel micropores can be formed in each hole of the micropore plate, so that the repeatability requirement of subsequent further experiments in which cell microspheres are formed is met, and the waste of cells, especially for precious patient-derived cells, is not caused. The number, the position and the size of cell microspheres formed by the hydrogel micropore array manufactured by using the die are controllable; moreover, multiple repeated experiments can be realized only by carrying out one-time treatment, such as drug treatment, in the holes of the cell culture microplate, so that the influence of the operation of an operator on an experiment result is reduced, and the experiment efficiency and accuracy are improved.

A method for producing extracellular vesicles (EV) in large quantities by using a peptide derived from Noxa protein, which plays a key role in apoptosis, and derivatives thereof, are described. Also described are the use of the peptide for promoting extracellular vesicle production and methods for producing extracellular vesicles by using the same. The methods of the present invention can efficiently and uniformly be used to mass-produce extracellular vesicles and can be used to produce extracellular vesicles which load drugs that pass poorly through cellular membranes, as well as recombinant proteins or plasmid DNA.

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.

Platelet lysate compositions and cell culture media compositions for maintaining and / or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and / or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and / or flasks pretreated with the cell culture media compositions.

Embodiments are described that relate to methods and systems for expanding cells in hollow fiber bioreactors. In embodiments, the cells are exposed to multiple growth factors, including combinations of recombinant growth factors. In other embodiments, the cells are grown in co-culture with other cells, e.g., hMSCs. In embodiments, the cells comprise CD34+ cells. The coated membrane comprises a membrane having a first coating configured to promote cell attachment to the membrane and a second coating comprising a soluble protein moiety.

The disclosure relates to growing cells, directing cells to grow into specified cell types, genetically and physically manipulating cells, and addressing one or more individual cells within a mixed cell population. Aspects of the disclosure relate to vectors useful to induce developmental changes in cells, in which those vectors have a temporal component. Vectors of the disclosure encode a controllable, temporal series of events. Once the vectors are delivered into target cells, a series of discrete and different genetic events may be induced. The disclosed methods generally provide for the temporal encoding of multiplex genetic effectors in vector format for cell state transitions.

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 methot 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.

Provided is a cell culture substrate comprising a substrate and a layer that covers at least a portion of the surface of the substrate, has a layer thickness of 5 to 2000 nm and contains a hydrophilic polymer, in which the hydrophilic polymer contains a phosphorylcholine group or a hydroxyl group, the cell culture substrate has a region (A) and a region (B) as mentioned below, and the bump height in a boundary region between the region (A) and the region (B) is 1 to 500 nm. (A) An island-like region which has cell adhesiveness and cell proliferating properties and has an area of 0.001 to 5 mm2. (B) A region which is located adjacent to the region (A) and does not have cell adhesiveness or cell proliferating properties.

Systems, devices, and methods for treating a nerve injury in a patient are provided. The system includes an extracellular matrix, a neutralizing element, and a reconstituting element. The extracellular matrix is configured to promote and / or sustain the growth of tissue and / or associated tissue properties proximate the nerve injury.

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.

A tissue graft for soft tissue repair or reconstruction comprising a sheet of a biopolymer-based matrix having a plurality of small perforations and a plurality of large perforations. The small perforations are sized to facilitate clotting and granulation tissue development within the perforations which, in turn, facilitates revascularization and cell repopulation in the patient. The large perforations are sized to reduce the occurrence of clotting and granulation tissue development within the perforations so that extravascular tissue fluids accumulating at the implant site can drain through the tissue graft. The large perforations enhance mammal tissue anchoring by permitting mammal tissue to compress into the perforations increasing mammal tissue contact area.

Provided herein are devices and methods for improved oxygenation of biological cells transplanted into a biological subject. The devices and methods may be used to promote vascularization to a transplantation site of the biological subject, and / or may be used to improve survival and functionality of cells contained within an encapsulation device.

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 provides A method of producing a decellularised extracellular matrix (ECM) scaffold of at least a portion of a lobular organ with no common artery, the method comprising: a) closing afferent blood vessels to substantially seal a target lobular organ or portion thereof with no common and / or major artery within a non-human donor or a dead / brain dead human donor; b) optionally: (i) cleaning coagulum and / or blood from at least a portion of the closed afferent blood vessels; and / or (ii) perfusing the organ or portion thereof to confirm closure of the afferent blood vessels; c) removing the sealed organ or portion thereof from the donor; and d) perfusing the sealed organ or portion thereof with detergent and enzymatic solutions to obtain the decellularised ECM scaffold. Methods for producing an artificial organ, and artificial organs produced by the methods 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.

The present invention provides a method for producing spheroids and cellular by-products of spheroids, such as extracellular vesicles. The method comprises providing a bioreactor having a cavity for culturing cells; inserting a cell into the cavity to form a spheroid; and perfusing a cell culture medium through the cavity to culture the spheroid. A porous scaffold may be soluble or insoluble, and the cells are inoculated into pores of the porous scaffold, where the cells aggregate in the pores of the porous scaffold to form spheroids. The bioreactor is continuously perfused for large scale spheroid production and enables collection of spheroids and / or extracellular vesicles or other components from cell secretory groups.

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.