69 results about "Cell seeding" patented technology

The invention relates to the technical field of biomedical engineering, in particular to a separated type organ-like chip model and a use method thereof.The model comprises a chip, an organ-like unit module and a fluid control system, and the use method comprises the steps that firstly, the chip is assembled and prepared; step 2, cell inoculation and culture; step 3, establishing and operating a fluid circulation system; step 4, organ-like unit function detection and data acquisition; 5, analyzing an experimental result and adjusting the model; the model is connected with a micro-fluidic channel through an independent cavity, and the characteristics, cell types and culture conditions of a cell culture bracket can be regulated and controlled according to the characteristics of simulated organs. Meanwhile, the micro-fluidic channel ensures stable and ordered material exchange among the organoid units, and is beneficial to more accurately screening out effective treatment schemes and drug targets.

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.

The present invention relates to a method for culturing cells, as well as a cell culture device and a kit used in the culturing method. The method for culturing cells of the present invention includes applying cells to a polyimide porous membrane and culturing them. In one embodiment of the method of the present invention, it includes a step of inoculating cells onto the surface of the above-mentioned polyimide porous membrane. Alternatively, in one embodiment, it includes the following steps: loading a cell suspension onto the dry surface of the above-mentioned polyimide porous membrane, placing the above-mentioned polyimide porous membrane, or moving the above-mentioned polyimide porous membrane to promote the outflow of liquid, or stimulating a part of the surface to suck the cell suspension into the above-mentioned membrane, and then accumulating the cells in the cell suspension into the above-mentioned membrane and allowing the water to flow out. Alternatively, in one embodiment, it includes the following steps: wetting one or both surfaces of the above-mentioned polyimide porous membrane with a cell culture solution or a sterilized liquid, loading a cell suspension onto the wetted polyimide porous membrane, and then accumulating the cells in the cell suspension into the above-mentioned membrane and allowing the water to flow out.

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 invention relates to the field of biomedical detection, and provides a novel chromosome analysis smear processing system and method. The system is based on a full-automatic sample treatment platform and comprises an automatic cell inoculation and culture module, an automatic cell harvesting module, an automatic cell dropping module and a cell genetic information management system, the single-batch treatment flux is not less than 192, and the system can be expanded. And precise control of each link is realized through technologies of a high-precision split charging pump, electric field assisted cell separation and the like. The method comprises the steps of automatic inoculation, constant-temperature culture, electric field assisted harvesting, precise dropping and data monitoring and analysis. Compared with a traditional mode, the method solves the problems of complicated equipment, low flux, insufficient standardization and the like, has the advantages of high processing flux, high standardization and informatization degree, small operation error, intelligent quality control and the like, and can effectively improve the cytogenetics analysis efficiency and accuracy.

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 invention relates to the technical field of cell biology, and discloses a chromosome aberration test method which comprises the following steps: inoculating CHL cells into a plate with the diameter of 100mm, and culturing in a carbon dioxide incubator at 37 DEG C for 24 hours; 4 hours before the cells are harvested, adding a colchicine solution to block the cells in a mitosis metaphase; adding 0.075 mol / L KCL solution for resuspending the cells, and putting the cells into a water bath of about 37 DEG C for hypotonic for 30 minutes to expand the cells; 5 min before the hypotonic treatment is finished, 1 mL of fixing liquid is added for pre-fixing, after uniform mixing, 200 G centrifugation is conducted for 5-7 min, and supernate is discarded; slowly adding 5mL of fresh stationary liquid, uniformly mixing, fixing in a water bath at about 37 DEG C for 30 minutes, reversing uniformly mixed cells every 10 minutes in the process to fully fix, centrifuging at 300G for 10 minutes, discarding supernate, and fixing twice by the same method; adding a set amount of stationary liquid to resuspend the cells, and fixing overnight at 4 DEG C; the chromosome aberration test method provided by the invention solves the technical problem that the chromosome aberration condition in the genetic toxicology test is not easy to observe in the prior art.

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。

The invention discloses an integrated cell culture and detection condition optimization method, which comprises the following steps of: establishing an oxygen consumption and culture solution pH value prediction model for cell culture in a closed system; the conditions such as cell inoculation amount, cell culture liquid volume, cell culture time and initial sodium bicarbonate addition amount required by closed system cell culture are optimized according to the model prediction trend, and the oxygen partial pressure in the gas phase of the closed headspace sample bottle is maintained at the physiological level of human tissues and organs within the culture time range; the pH value of the culture solution is maintained near the physiological level (pH = 7.4), the growth curve of the cells cultured under the closed system under the optimized culture condition is consistent with that of the cells cultured under the traditional air-permeable cell culture, the method provided by the invention can realize in-situ detection of the cell growth activity in the closed system, realizes integrated cell culture and detection, does not need to damage the closed state of the cells, and is suitable for large-scale production. The cell culture environment is free of pollution risk and steady-state damage.

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 invention relates to the field of biomedical engineering. The invention discloses a method for enhancing ToF-SIMS detection sensitivity in a single cell. The method comprises the following steps: S1, carrying out ITO glass surface treatment, preparing a nano capillary tube, and representing the nano capillary tube; s2, inoculating the MCF-7 cells to ITO glass, and culturing the MCF-7 cells; s3, carrying out treatment on the MCF-7 cells; s4, injecting the quantum dot solution into the MCF-7 cells under the action of voltage; s5, cleaning and freezing the ITO glass; and (S6) analyzing the MCF-7 cells by using ToF-SIMS (Tooth of Free- The solution at the tip of the nanocapillary drifts by applying voltage, so that the control on the liquid at the tip of the nanocapillary is realized. By using the electrochemical injection method, the ultramicro-volume matrix molecules and the nanometer material solution are injected into the cells, so that secondary ion signals of amiodarone and biomolecules in the MCF-7 cells are enhanced.

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.

A cell culture substrate is provided having a substrate having 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 through the thickness of the substrate. The plurality of openings permit at least one of cell culture media, cells, or cell products to flow through the thickness of the substrate and provide a uniform, effective, and expandable matrix for cell inoculation, proliferation, and culture. The substrate may be formed from a woven polymer web material that provides a high surface area to volume ratio for cells, as well as excellent fluid flow through the matrix. Bioreactor systems comprising the cell culture matrices and related methods are also provided.

The present invention relates to the technical field of cell culture, and specifically discloses a method for domesticating an adherent cell line into a fully suspended cell line. The adherent-dependent cells are inoculated on microcarriers and cell proliferation is carried out in DMEM medium containing 10% fetal bovine serum. After the cells proliferate to 10E6 cells / mL, the microcarriers are lysed, the cell clusters are centrifuged and collected. The cell clusters are inoculated in a medium containing 10% fetal bovine serum, 0.1% shear force protectant, and 2 g / L glucose and cultured at a rotation speed of 30 to 50 rpm for 3 to 4 days to enable the cells to form embryoid body clusters of 50 μm to 150 μm. The embryoid body clusters are mechanically pipetted to disperse them, and are domesticated for 18 to 22 days with DMEM medium containing different concentration gradients of low molecular weight dextran sulfate to obtain a successfully suspended and domesticated cell line. The method of the present invention improves the uniformity of cell embryoid bodies, makes the properties of the embryoid bodies more stable, and can efficiently obtain a suspended cell line.

The utility model provides a disposable adjustable device for a cell scratch experiment. The cell culture plate scratching device comprises scratching arms, fixing cross rods and vertical rods, the specification of the scratching arms is matched with the diameter of the bottom face of a culture hole of a cell culture plate to be placed, a plurality of sawtooth structures distributed at preset intervals are arranged on the upper surfaces of the scratching arms, each folding arm is connected with the corresponding vertical rod, the vertical rods are connected through the fixing cross rods, and the fixing cross rods are connected with the scratching arms. A lifting partition convenient for a worker to lift is arranged above the fixed cross rod, a dismounting crease is arranged at the bottom of the fixed cross rod, and the length of the fixed cross rod is subjected to preset breaking adjustment based on the specifications of different to-be-placed cell culture plates. According to the utility model, the length of the scratching arm and the height of the fixed cross rod can be adjusted according to the depth and the diameter of the culture hole, so that the scratching arm is attached to the bottom of the culture hole, then cells are inoculated on two sides of the scratching arm, and the phenomena of scratching deviation and the like caused by improper operation are avoided.

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.