47 results about "Cell seeding" patented technology

The utility model belongs to the technical field of cell culture, and particularly relates to an automatic cell collecting device which comprises a rack and a sealing cover arranged on the rack, a normally closed butt joint window is arranged on one side of the sealing cover, and the butt joint window can be connected with a culture box; a culture dish can be placed in the culture box; the first mechanical arm is arranged on the operation platform; the liquid collecting mechanism is arranged on the operation platform, the liquid input mechanism is arranged on the operation platform, a first cover opening assembly is further arranged beside the liquid input mechanism, and the centrifugal collecting mechanism is arranged on the operation platform. By arranging the butt joint window, the first mechanical arm, the liquid collection mechanism, the liquid input mechanism and the centrifugal collection mechanism, the cell collection process can be fully automatically completed, the automation degree is high, the risk of artificial secondary pollution is reduced, meanwhile, a production line can be synthesized with a cell inoculation device, the first mechanical arm is shared, the size of the machine is reduced, and the production cost is reduced. The occupied area is small, the cost is low, and the structure is simpler.

The invention discloses a method for culturing NK (Natural Killer) cells. The method comprises the following steps: inoculating a mononuclear cell into a T25 cell culture flask, wherein the T25 cell culture flask is coated by a coating factor in advance; carrying out culture treatment on the inoculated mononuclear cells in a culture system containing an NK cell culture medium containing an activation amplification factor and plasma so as to obtain NK cells; wherein the coating factor, the activation amplification factor and the NK cell culture medium are derived from an NK cell amplification kit. According to the method provided by the embodiment of the invention, the dosage of the amplification reagent can be reduced, the amplification multiple of the NK cells is improved, and the production cost is reduced.

A cell culture device according to the present disclosure comprises: a culture chamber for culturing suspension cells; a flow path having a culture medium supply port and a culture medium discharge port, and provided adjacent to part or all of the outer side of at least one surface of the culture chamber; an oxygen permeable membrane provided to at least part of the flow path and / or the culture chamber; and a cell seeding port provided to at least part of the culture chamber.

The present disclosure provides devices, instruments, and kits for handling certain implantable substrates, for example, cell-seeded membranes. The devices, instruments, and kits may be used for washing, cutting, and preparing the substrates for implantation into a patient. The devices, instruments, and kits may be particularly useful for handling implantable substrates that were cryopreserved.

The invention discloses an imaging analysis method adopting a dome microscope light source. Relates to the technical field of microscopes. The method comprises the following steps: inoculating cells into micropores of a cell culture plate; placing the cell culture plate below a dome microscope light source, focusing, moving the cell culture plate after the focusing is clear, and enabling the field of view of the dome microscope to be at the edge of the micropore of the cell culture plate; turning on a middle light source in the light source inner ring of the dome microscope, adjusting the brightness of the middle light source and the dome light source, and obtaining a clear cell culture plate edge image by automatically adjusting the brightness and automatically analyzing the collected image; and splicing a plurality of images to obtain a complete and clear image of the whole micropore. According to the invention, the problem of imaging shadow caused by the concave liquid surface effect in the traditional inverted microscopic imaging experiment is effectively solved. The light of the light source can better penetrate through the concave liquid surface and uniformly irradiate each part of the microwell plate, so that the imaging of the edge of the microwell does not have a shadow any more, and the quality and definition of the imaging are improved.

A cell culture matrix is provided that has a substrate with a first side, a second side opposite the first side, a thickness separating the first side and the second side, and a plurality of openings formed in the substrate and passing through the thickness of the substrate. The plurality of openings allow flow of at least one of cell culture media, cells, or cell products through the thickness of the substrate, and provides a uniform, efficient, and scalable matrix for cell seeding, proliferation, and culturing. The substrate can be formed from a woven polymer mesh material that provides a high surface area to volume ratio for cells and good fluid flow through the matrix. Bioreactor systems incorporating the cell culture matrix and related methods are also provided.

The application discloses a kind of clear supply seed synchronous cell seed metering device, belong to the technical field of agricultural equipment.The device includes front and rear shell, disc type rotating seed cavity, guide groove type seed metering disc and surplus seed partition recovery mechanism.Through the intermittent rotation of disc type rotating seed cavity, the synchronization of seed supply and cleaning is realized;Seed metering disc is disturbed and orderly guided under the action of negative pressure, and the seed population dispersity and single seed rate are improved;After completing seeding, surplus seeds are transported to the partition recovery device by negative pressure airflow, and independent storage of surplus seeds in each cell is realized using the grid structure, and rotation according to the grid is realized by servo drive, to avoid seed mixing.The device has simple structure, reliable operation, can effectively improve the cell seeding efficiency and seeding quality, and meet the demand of surplus seed traceability management in breeding process.

Described herein are systems, methods, and interconnecting porous hydrogel blocks for co-cultivation of a plurality of cell types. A block may comprise a 3D continuous polymeric matrix with a network of microporous cavities, and may be configured to interconnect with at least one other block. A system may comprise at least one block and be configured to co- cultivate the cell types when seeded with at least one cell of each type. A method may comprise seeding at least one block with at least one cell of each type.

本发明公开了一种CD34+造血干细胞化学重编程为诱导多能干细胞的方法。所述方法包括：将CD34+造血干细胞分散为单细胞，接种于包被细胞外基质的培养表面，在无血清培养基中依次进行三阶段诱导培养：第一阶段培养基包含丙戊酸、CHIR99021、RepSox、EPZ6438、PY60、OAC1和OAC2；第二阶段培养基包含丙戊酸、CHIR99021、RepSox、反苯环丙胺、TTNPB、Forskolin、OAC1和OAC2；第三阶段培养基包含CHIR99021、Forskolin、Y‑27632、SP600125、OAC1和OAC2。最后挑取形成的iPSC克隆进行扩增培养，获得诱导多能干细胞。本发明针对CD34+造血干细胞优化了小分子组合和诱导时序，全程无病毒、无转录因子、无血清，重编程效率可达3%以上，细胞凋亡率低，可获得具有完整多能性和正常核型的iPSC，适用于临床级细胞制备和再生医学应用。

This application discloses a method for capturing chromatin conformation, comprising: S1, seeding target cells into multiple wells; S2, performing chromatin conformation capture processing on the target cells in each well in S1 to obtain target gene fragments; S3, fragmenting and labeling the target gene fragments so that each target gene fragment in each well is labeled with a unique barcode sequence, so as to analyze the chromatin three-dimensional structure of the target cells in each well. The method of this application can efficiently and on a large scale perform chromatin conformation capture and data processing, which is beneficial for the discovery of chromatin three-dimensional structure remodeling factors and the research on the validation of druggable target genes.

Provided herein are methods of producing and manufacturing iPSCs, such as iPSCs essentially free of exogeneous viral residuals, and RPE cells. Such methods may comprise cell seeding at low density (e.g., single-cell passaging) and / or culturing at elevated temperature.

The utility model discloses automatic production equipment for cell drugs. The automatic production equipment comprises a process operation cavity and a cell factory incubator, the cell factory incubator is communicated with the process operation cavity; the process operation cavity is provided with a cell harvesting area and a cell inoculation liquid changing area; a centrifugal bottle pipetting and cover opening and closing assembly, a centrifugal assembly, a subpackaging assembly and a first transfer mechanism are arranged in the cell harvesting area; the first transfer mechanism is used for transferring the centrifugal bottle between the centrifugal bottle pipetting and cover opening and closing assembly and the centrifugal assembly; a cell factory pipetting and cover opening and closing assembly and a second transfer mechanism are arranged in the cell inoculation liquid changing area; the second transfer mechanism is used for transferring the cell factory among the cell factory pipetting and cover opening and closing assembly, the cell factory incubator and the centrifugal bottle pipetting and cover opening and closing assembly. By means of the arrangement, the function of full-process automation is achieved, and the purposes of reducing manual intervention and improving operation efficiency and sterility are achieved.

The invention belongs to the technical field of cell culture devices, and particularly relates to a device for in-vitro construction of a 3D neural immune cell co-culture model.The device comprises a vessel body container, a vessel cover is arranged outside the vessel body container, a biological scaffold is arranged in the vessel body container, and the biological scaffold comprises a static column, a plurality of movable plates and a plurality of half grooves; the plurality of movable plates are arranged around the static column and can rotate by taking the static column as an axis, the plurality of movable plates divide the vessel body container into a plurality of culture chambers, and each movable plate is provided with a plurality of gaps, so that the plurality of culture chambers are communicated through the gaps; half grooves are formed in the opposite positions of every two adjacent movable plates; and when two adjacent movable plates are folded, the corresponding half grooves are spliced to form a complete cell inoculation groove. The device is good in stability, different culture chambers are arranged in the vessel body container, materials cannot be leaked to the outside of the vessel body container, pollution is prevented, and safety is high.

Embodiments of methods, systems, and devices are disclosed for screening of cells for chirality. The methods include seeding and culturing cells onto a substrate having microgrooves, collecting images of the cells on the substrate, and performing an image analysis to determine the angular distribution of the cells with respect to the microgrooves. For some embodiments, the image analysis involves the use of fast Fourier transforms to determine the angular distribution. The methods, systems and devices provide improved means for rapidly evaluating cellular chirality.

A cell seeding apparatus of the present disclosure includes a dropping device that drops a liquid droplet of a cell suspension into a cell culture vessel having a measurement electrode and a reference electrode disposed on a bottom surface, an imaging device that is provided in the dropping device and that images the bottom surface to generate an image, and an information processing device that detects positions of the measurement electrode and the reference electrode based on the image and that decides, based on detection information, a dropping position of the liquid droplet at which cells are to be placed on the measurement electrode without placing the cells on the reference electrode.

The estimation device (200) generates quality prediction data (540) indicating the quality of a drug substance of a biopharmaceutical manufactured by culturing the cells, by inputting assay data (510) containing an assay result obtained by assaying the substance in the culture vessel at at least one timing after a prescribed period has elapsed from inoculation of the cells into the culture medium, to a prediction model (420). The prediction model (420) is generated by performing a learning process using learning data (530) containing assay data containing assay results obtained by assaying the substance in the culture vessel at a plurality of timings from inoculation of the cells into the culture medium, and quality data obtained by analyzing the drug substance of the biopharmaceutical manufactured with the cells.

It has been determined that optimizing cell seeding on tissue engineered vascular grafts (TEVGs) is associated with reduced inflammatory response and reduced post-operative stenosis of the TEVG. Cell seeding increases TEVG patency in a dose-dependent manner, and the more cells seeded, the better the TEVG patency, however, incubation duration exhibits minimal effect on TEVG patency. Methods of engineering patient-specific TEVGs are provided, including optimal cell numbers for maintaining graft patency and reducing post-operative stenosis. A closed, disposable customizable system for seeding TEVGs is also provided. To enhance the efficacy of cell seeding, the system is preferably custom designed based on the morphology of the patient-specific graft.

The present disclosure provides devices, instruments, and kits for handling certain implantable substrates, for example, cell-seeded membranes. The devices, instruments, and kits may be used for washing, cutting, and preparing the substrates for implantation into a patient. The devices, instruments, and kits may be particularly useful for handling implantable substrates that were cryopreserved.

The invention relates to the technical field of micro-fluidic organ chips, and discloses a micro-fluidic testis organ chip and a culture method, the micro-fluidic testis organ chip comprises a PDMS fluid control layer and a PDMS cell culture layer from top to bottom; the at least three culture units are arranged on the PDMS cell culture layer in the axial direction, each culture unit internally comprises three functional areas, the three functional areas are used for testicular sertoli cell culture, spermatogenic cell culture and basement membrane simulation respectively, and the three functional areas are communicated with one another to form a fluid path; the regulation and control part is arranged on the PDMS fluid control layer, and the regulation and control part is used for regulating and controlling fluid paths which are communicated with one another among the three functional areas; the inoculation and collection part is arranged on the PDMS fluid control layer and is used for carrying out cell inoculation and collection in the three functional areas. The in-vivo testis spermatogenesis microenvironment is highly simulated, the limitation of a traditional 2D cell experiment and an animal experiment can be effectively made up, and a new method and platform are provided for analyzing a spermatogenesis disorder mechanism and screening targeted repair drugs.

The invention discloses an ozone gas-liquid cell contamination method based on a Transwell chamber, and belongs to the technical field of in-vitro gas contamination. The invention provides an O3 gas-liquid exposure mode based on a Transwell chamber by improving the contamination mode, and achieves the effects of simplifying the experimental method, reducing the experimental cost and improving the contamination efficiency. The gas-liquid exposed interface is constructed by utilizing the Transwell chamber, and cell inoculation, culture and sampling methods are simple and convenient; the Transwell chamber and the cell culture box with the O3 generation and real-time concentration monitoring functions are simple in principle, easy to understand and easy to obtain, and the O3 concentration can be monitored in real time; the experimental device is small in occupied space, low in building cost, good in operability and high in practicability.

A method of introducing a substance into a cell includes a) seeding floating cells on a gel plate medium having a specific hardness, b) filling the gel plate medium seeded with cells with an electrolytic solution and positioning a nanopipette filled with a substance at a cell-corresponding position in the electrolytic solution, c) measuring a current between an inside of the nanopipette and the electrolytic solution and moving the nanopipette in a cell direction to a position at which a drop rate from a steady state current is a set current drop rate, d) moving the nanopipette in the cell direction by a set penetration distance to puncture a cell, e) applying a voltage between the inside of the nanopipette and the electrolytic solution to eject the substance into the cell, and f) removing the nanopipette.

The utility model relates to the field of cell culture, in particular to an organ chip which comprises a bottom plate, a culture tank detachably connected to the bottom plate and a protective cover used for protecting a liquid flow channel and the culture tank. The culture tank comprises a containing part and a microporous membrane. The accommodating part is cylindrical, one end of the accommodating part is open, and the other end is closed by a microporous membrane; the bottom plate is provided with a liquid flow channel used for containing a culture medium. The liquid flow channel is provided with a matching part and a communicating part penetrating through the two sides of the matching part. The matching part is used for accommodating and constraining the culture tank, and the communicating part is used for adding and taking liquid into and out of the liquid flow channel. And when the culture tank is detachably sleeved on the inner side of the matching part, the culture tank is separated from the bottom of the liquid flow channel. The protective cover detachably covers the bottom plate. Through the detachable design, the difficulty of model sampling and cell inoculation is reduced, dynamic culture can be realized through shaking of the communication part, the operation difficulty and cost are reduced, and the practicability is improved.

The invention discloses a whole genome open chromatin in-situ labeling method and application thereof in detection of whole genome open chromatin spatial distribution, and belongs to the technical field of cell molecular markers. The method comprises the following steps: (1) preparing a Tn5-fluorescent probe transposon storage solution; (2) taking cells, inoculating the cells on a carrier for division growth, cleaning with a buffer solution, fixing, cleaning with the buffer solution, treating with a lysis buffer solution, and cleaning with the buffer solution; (3) buckling the carrier with the cell side treated in the step (2) on a sealing film containing a standard reaction system, and incubating in a sealed wet box; and (4) taking out the incubated carrier with the cells, and washing the carrier with a washing buffer solution to obtain the Tn5-fluorescent dye transposon in-situ labeled cells. The labeling method disclosed by the invention can be used for carrying out fluorescence labeling on open chromatin regions of genomes in different types of cells, and has extremely high sensitivity and short time consumption.

The present disclosure describes methods of maintaining the phenotype of differentiated cells. Generally, the natural environment of the body is replicated for the differentiated cell. The differentiated cell is plated on a cell culture substrate comprising a laminin, such as laminin-521 or laminin-511. The substrate may also contain a cadherin. This maintains the phenotype of the differentiated cell.

The utility model belongs to the technical field of cell culture, and particularly relates to an automatic cell inoculation device which comprises a rack and a sealing cover arranged on the rack, a closed operation space is formed by the sealing cover and the top side of the rack, an operation platform is arranged in the operation space, one side of the sealing cover is provided with a normally-closed butt joint window, and the other side of the sealing cover is provided with a movable butt joint window. The butt joint window can be connected with an incubator; a culture dish can be placed in the culture box; the mechanical arm is arranged on the operation platform, and the mechanical arm can carry a culture dish; the conveying assembly is arranged on the operation platform, the inoculation assembly is arranged on the operation platform, and the inoculation assembly comprises a first uncovering assembly, a culture medium input assembly and a liquid transferring assembly which are arranged beside the conveying assembly. By arranging the butt joint window, the mechanical arm, the transportation assembly and the inoculation assembly, the cell inoculation process can be fully automatically completed, the automation degree is high, and the risk of artificial secondary pollution is reduced.

The present invention relates to a cell culture device suitable as an organ-on-chip device. The device is characterized by having two fluid circuits, one to allow seeding a culture chamber with cells, and a second circuit to maintain a continuous feed of the culture medium until the seeded cells have a suitable degree of growth. Particularly, the device is adapted for cell culture to be carried out in a hydrogel matrix that allows simulation, for example, of the development of tumors in a biological organ.

This disclosure relates to applications of cell culture, and more specifically to applications of cell culture for growing cells such as stem cells (e.g., PSCs). The method comprises seeding cells in a first container and optionally transferring the cells from the first container to a second container. The cells in the first container, and, if applicable, the cells in the second container, are brought into contact with at least one externally added extracellular matrix or extracellular matrix protein. The method of this disclosure yields a population of cells that are significantly and rapidly grown while maintaining cell quality. The method of this disclosure may also improve the cell seeding efficiency of cells that are significantly and rapidly cultured / grown in a monolayer.

A device for cell seeding according to an exemplary embodiment includes a first layer and a second layer. The first layer includes a first accommodation space configured to accommodate a cell to be seeded. The second layer includes a first through-hole wider than the cell and is positioned vertically below the first layer. Then, the device for cell seeding forms a first state in which the first through-hole is positioned at a bottom surface of the first accommodation space by changing a relative position between the first layer and the second layer in a horizontal direction.

To prevent foam buildup and ensure uniform cell seeding, the uneven distribution of flow velocity in the width direction of the channel is reduced in curved channels. [Solution] The cell culture chip of the embodiment is a cell culture chip that includes a laminate having a flow channel structure inside, wherein the laminate includes a flow channel substrate having a flow channel leading to a port, the flow channel includes a curved flow channel formed in a curved shape in a plan view of the flow channel substrate and having a cross-sectional area that gradually changes in the direction of fluid flow, and a first connecting flow channel that connects the port and the curved flow channel and has a constant cross-sectional area in the direction of fluid flow, wherein the cross-sectional area of ​​the first connecting flow channel is smaller than the minimum value of the cross-sectional area of ​​the port and the cross-sectional area of ​​the curved flow channel, respectively, and in the plan view, the flow channel width of the curved flow channel gradually changes in the direction of fluid flow.

A method is provided for producing a cell inoculum suitable for use in inoculating a bioreactor in a biomanufacturing or cell culture process. The method includes the steps of taking a cell line sample and undertaking at least one cell passage with said cell line sample to increase the number of cells to form at least a first set of passaged cells. Subsequently said first set and / or one or more subsequent sets of passaged cells is used for optionally inoculating a bioreactor in use. The method further includes the step of applying electrical or electromagnetic stimulus, wave and / or one or more signals to any or any combination of: the cell line sample, the at least first set of passaged cells and / or one or more subsequent sets of passaged cells to create a pre-conditioned cell inoculum. Optionally the pre-conditioned cell inoculum is then used to directly or indirectly inoculate the bioreactor.A cell inoculum composition suitable for use in a biomanufacturing or cell culture process and comprising a population of epigenetically modified cells is also provided. The cells may, for example, be HEK cells or CHO cells.