1411results 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.

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.

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 cell and tissue sheet forming package includes a container body, a membrane, a sliding door plate and a sealing film. The sliding door plate is disposed slidably on a top of the container body to cover or expose the membrane. The sliding door plate has a hole and a passive magnetic assembly. The cell injection equipment includes a carrier, an injection mechanism and a drive mechanism. The carrier carries the package, and the drive mechanism moves the carrier and the injection mechanism to have the injection mechanism to inject a solution, through the hole, into the package. A heating element of the carrier is introduced to heat the membrane and the solution to transform the solution into a colloid sheet on the membrane. Then, the positive magnetic assembly engages magnetically the passive magnetic assembly to slide the sliding door plate to expose the colloid sheet on the membrane.

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.

The disclosure provides synthetic cytotoxic cells, comprising a hydrogel microsphere and a TNF-related apoptosis-inducing ligand (TRAIL), or active fragment thereof. In aspects, the TRAIL or active fragment thereof is attached to the surface of the hydrogel microsphere. The disclosure further provides methods of preparing the synthetic cytotoxic cells and methods of use thereof in the treatment of cancer.

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.

This invention is intended to enhance production of viral vectors when producing viral vectors using viral vector-producing cells. A protein involved in the DNA damage response in viral vector-producing cells is inhibited.

Provided is a cell culture scaffold including a scaffold; and a cell culture coating layer which covers at least a portion of the surface of the scaffold and comprises a fusion polypeptide including a peptide for cell culture and a peptide for adhesion. Accordingly, the cell culture scaffold can achieve high cell culture efficiency due to having excellent adhesiveness with respect to various cells and being capable of stably proliferating adhered cells, can be stored at room temperature for several years, thus having excellent storage stability, prevents imaging interference, which can occur during cell observation or fluorescence analysis on the cell culture scaffold, due to the smooth surface of the cell culture coating layer formed by the fusion polypeptide, thus being applicable to various cell cultures, and, in particular, can express an excellent effect on the proliferation and differentiation of neural stem cells or neural precursor cells.

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.

A graft method according to the present invention comprises: a culture step in which a cell sheet is cultured on a culture surface of a culture carrier substrate; and a graft step in which the cell sheet formed on the culture surface of the culture carrier substrate is attached to a recipient site, after which the culture carrier substrate is detached from the cell sheet.

The present disclosure provides feeder hydrogel particles that can function to support the growth, proliferation, and / or activation of a target cell in culture. The present disclosure also provides methods of culturing target cells with feeder hydrogel particles.

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.

Provided herein is a see-through transparent, stable, safe and (bio)degradable hydrogel-based particulate support medium, made of calcium alginate particles. The calcium alginate particles, or hybrid hydrogel particles, are characterized by a substantially homogeneous average particle size that ranges from 0.1 micrometer to 5 micrometer.

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.

A tissue regeneration promoting sheet (10) is provided with a cell holding sheet (12) which is a non-woven fabric that contains fibers mainly composed of polycaprolactone and is capable of allowing a cell suspension containing stem cells to permeate therethrough. The fiber diameter of the fibers constituting the non-woven fabric may be in the range of 300 nm to 2 [mu] m. The weight per unit area of the non-woven fabric may be in the range of 1 g / m2 to 3 g / m2. The thickness of the non-woven fabric may be 300 [mu] m or more. The tissue regeneration promoting sheet (10) may also be provided with a reinforcing sheet (14) which is laminated on the nonwoven fabric and into which the cell suspension can permeate.

Systems, devices, and methods for producing a tissue specimen and performing subsequent biomechanical testing thereof to quantify various biological / physiological properties and characteristics. The system may include a bioreactor device for producing the tissue specimen which includes a packaged / sealed seeding cup containing a scaffold in a sterile environment amenable to seeding with cells. After seeding, the cup may be stored in a standard laboratory culture tray for incubation. The produced tissue specimen may next be transferred to a testing instrument under control of a microcontroller which automatically performs a series of tests on the tissue and generates testing data communicated to an external electronic device which can operably interact with and control operation in part of the testing instrument via software. Tissue integrity in the instrument may be maintained by sterile fluid circulation and heating systems. The system advantageously provides efficient and reproducible tissue growth and analysis.

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.