31 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 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.

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.

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.

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.

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 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 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.

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.

The invention provides a casein source 3D printing bio-ink and a preparation method of a biological scaffold and cell culture meat. The bio-ink is prepared from the following raw materials in parts by weight: 15.6 parts of gelatin; 2.6 parts of sodium alginate; 4.2 parts of casein; 52.5 parts of a sodium hydroxide deionized water solution with a pH value of 8; and 87.5 parts of a sterile 1 * PBS buffer solution. The ink is subjected to extrusion type 3D printing and calcium chloride crosslinking to obtain a biological scaffold; c2C12 cells are inoculated on the support, and cell culture meat is obtained through proliferation-differentiation culture. The problems that existing 3D printing bio-ink is single in performance and poor in component adaptability, a biological scaffold material is insufficient in mechanical stability, and nutritional ingredients are deficient are solved. The prepared bio-ink has excellent mechanical properties and printing adaptability, and provides key technical support for industrial preparation of cell culture meat.

This invention discloses a live-cell drug screening method and system based on FRET two-hybrid analysis. Firstly, preliminary experiments are conducted on multi-well plates using cell samples transfected with FRET standard plasmids to determine experimental conditions, including cell seeding density, imaging objective, excitation light intensity, field of view analysis range, and measurement of FRET system calibration parameters. Secondly, live-cell drug screening based on FRET two-hybrid analysis is performed under the preliminary experimental conditions. Cell culture is first performed, and drug and control groups are set up, followed by FRET three-channel imaging to calculate E0. D E A and R c And draw E D -R c Figure and E A -(1 / R) c (Figure); then use E D -R c With E A -(1 / R) c The linearly separated FRET two-hybrid analysis method yielded E Dmax E Amax and N A / N D Data was screened using the saturation index sRatio; finally, the active and potential active substances in the drug group were obtained through a significant difference test. This application enables accurate and rapid quantitative FRET measurement under a wide-field microscope with low magnification, meeting high-throughput requirements and screening for active compounds in live cells.

The invention discloses an identification method and application of an intercellular tunnel nanotube and a transport substance thereof, and belongs to the technical field of biological identifier.The identification method of the tunnel nanotube in a fixed state comprises the following steps that a culture dish is inoculated with cells to be detected, and after fixation, permeabilization and F-actin dyeing are conducted, the cells to be detected are placed in the culture dish; performing three-dimensional imaging to obtain a three-dimensional image; based on a preset identification standard, analyzing the three-dimensional image to identify the intercellular tunnel nanotube; the method for identifying the tunnel nanotube in the living cell state and the transported substance of the tunnel nanotube in the living cell state comprises the following steps: inoculating a to-be-detected cell in the living cell state on a culture dish, carrying out living cell imaging based on a differential interference comparison system, and accurately judging the key identification key point of the tunnel nanotube, namely whether the tunnel nanotube is attached to the bottom of the culture dish or not. The method can accurately and efficiently identify and distinguish tunnel nanotubes between cells with complex forms in fixed and living cell states.

Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), for example.

This invention discloses a highly efficient method for separating primary cells from cochlear tissue based on physical filtration. The method includes: S1: tissue pretreatment and viability preservation; S2: controlled mechanical dissociation; S3: automatic cell-impurity separation; b. differential centrifugation using a double-layer filtration centrifuge device; S4: impurity and cell removal and cell purification; S5: primary cell seeding and optimized culture. This separation method utilizes the natural differences in volume and mass between cells and bone fragments through a double-layer filtration centrifuge device and differential centrifugation filtration, simultaneously performing centrifugation sedimentation and filtration separation. Compared to traditional manual filtration or density gradient centrifugation, this device is simple to operate, has a fast separation speed, fundamentally avoids filter membrane clogging, and significantly improves separation efficiency, enabling the gentle and efficient separation of highly active primary cells from cochlear tissue.

The invention discloses a method for three-dimensional multiplication culture of a hepatic stem cell line. The method comprises the following steps: inoculating cells of the liver stem cell line into a low-adsorption culture plate containing a HepSC culture medium, and culturing at the rotating speed of 100-180 rpm to enable the cells to form cell spheres. According to the invention, the dependence of the HepSC cell line on mouse matrigel and adherent culture is effectively solved, a new three-dimensional culture way without matrigel is provided for large-scale amplification, and clinical transformation of the liver stem cell line can be promoted.

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.

An approach for forming organized 3D single-cell networks where their real-time signaling responses to a range of stimuli can be accurately captured using simple cell seeding and easy-to-handle microfluidic chips. Crosslinked collagen within multi-chambered microfluidic chips is formed, followed by femtosecond laser ablation of 3D microchannel networks and cell seeding. Cells migrate within ablated networks within hours, self-organize and form viable, interconnected, 3D networks in custom architectures such as square grid, concentric circle, parallel lines, and spiral patterns. The functionality of cell networks can be studied by monitoring the real-time calcium signaling response of individual cells and signal propagation when subjected to flow stimulus alone or a sequential combination of flow and biochemical stimuli. Furthermore, user-defined disrupted networks can be generated by lethally injuring target cells within the 3D network and analyzing the changes in their signaling dynamics.

Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), helper, naïve, memory, or effector, for example.

A cell culture vessel comprises an upper portion 12 comprising a volume for receiving a cell culture medium and cells, and a lower portion 20 in fluid communication 24 with the upper portion. Defined

The utility model provides a kind of cell culture device, cell culture device includes culture plate and cover plate, culture plate is equipped with multiple hole grooves of being arranged with mutual spacing;At least one of multiple hole grooves is provided with culture assembly, culture assembly includes multiple culture tanks of being arranged with mutual spacing;Culture tank is used to accommodate pre-cultured cell;Cover plate is detachably placed on culture plate, cover plate is equipped with multiple surround frame, multiple surround frame corresponds with multiple hole grooves.The technical problem solved by the utility model is that after cell inoculation is completed, it is difficult to maintain stable moving state during movement, resulting in the movement of cell mass distributed in the same hole, easy to appear mutual fusion, and then leading to cell overlap, uneven distribution and indirectly promoting side differentiation, affecting the pluripotency of stem cells.