121 results about "Adherent cell" patented technology

Provided herein are altered cell lines comprising a suspension cell line adapted from an adherent cell line having a different expression profile from a corresponding non-altered adherent cell line, methods for generating altered cell lines, and methods of characterizing altered expression profiles for a gene, a transcript, or a protein in an altered cell line.

The invention relates to the technical field of biology, and particularly discloses a device and a method for separating an adherent cell through a vortex sound field, an ultrasonic module, piezoelectric ceramics and an Archimedes spiral structure concave lens are arranged to generate the vortex sound field to act on a cell culture dish, so that an adjustable shear flow is generated on the cell culture dish, and the adherent cell is separated from the cell culture dish. And locally separating the adherent cells in the cell culture dish. According to the concave lens with the Archimedes spiral structure, the problem of defocusing of a high-order vortex sound field can be solved, and the falling control accuracy of adherent cells is improved. The proposed method does not require any physical contact with adherent cells, and does not require insertion of an instrument into a cell culture dish, thereby reducing the risk of contamination and cell damage. The stable flow generated by absorbing sound energy by the liquid is used for indirectly separating the adherent cells, the method is extremely low in energy density and has good biocompatibility, and the activity of the cells is ensured while the cells are efficiently separated.

The invention relates to a Vero cell serum-free culture medium and application thereof, the serum-free culture medium is suitable for free suspension growth of Vero cells and is free of animal sources and protein components, adherent cells can be directly domesticated through a certain method to screen out Vero suspension cells, and serum-free suspension culture of the Vero suspension cells is achieved in the serum-free culture medium. The density of cells subjected to suspension culture by adopting the serum-free culture medium can reach more than 107 cells / mL, the cell viability is more than 90%, the cells are well dispersed, the cells can still normally grow and proliferate after continuous subculture for 20 generations, and the cells are sensitive to various viruses.

The invention discloses a detection method for rapid real-time drug effect evaluation and application of the detection method. The detection method comprises the following steps: providing a quantitative corresponding relation between a same-generation calibration change value of electrical characteristics caused by physiological changes such as cell same-representative characteristics and the like and the cell proliferation ability; adhering to-be-detected cells to the surface of the same membrane electrode; applying the medicine to be detected, and observing the same generation measurement change value of the electrical characteristics in a cell proliferation cycle; and based on the quantitative corresponding relationship, obtaining the drug effect of the corresponding real-time proliferation characteristic evaluation drug. According to the detection method provided by the invention, the physiological changes such as the same-representative characteristics of the adherent cells are monitored through the membrane electrode, and after the quantitative corresponding relation between the change indexes of the same-representative characteristics of the cells and the proliferation capacity of the cells is established, the cell proliferation capacity is detected; the change of the cell proliferation capacity can be predicted through physiological changes such as the same representative characteristics of the cells before proliferation and apoptosis of the cells, so that the judgment period of the influence of the medicine on the cells can be greatly shortened, and the progress of medicine evaluation and research and development can be remarkably accelerated.

The invention discloses a cat kidney suspension culture cell line suitable for serum-free suspension culture and application of the cat kidney suspension culture cell line. The name of the cell line is F81-B4, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.46350. After 50 generations of continuous subculture, the cells still can maintain the original proliferation level and cellular morphology, the cell number can reach 9.0 * 10 < 6 > / mL after the cells are subcultured at the density of 1.0 * 10 < 6 > / mL for three days, the multiplication time is 22-26 hours, and the cell proliferation speed is equivalent to that in a shake flask when the cells are amplified to a 10L bioreactor for culture. The cell line is highly susceptible to feline parvovirus, feline calicivirus, feline herpes virus, canine parvovirus and canine distemper virus, the virus content of a virus solution harvested 72 h after FPV virus inoculation can reach 107.5 TCID50 / mL, the virus content of a virus solution harvested 24 h after FCV virus inoculation can reach 1010.8 TCID50 / mL, the virus content of a virus solution harvested 48 h after FHV-1 virus inoculation can reach 107.5 TCID50 / mL, compared with an existing F81 adherent cell production process, the production process has the advantages that the production efficiency is high, and the production cost is low. The method has the characteristics of rapid lesion, short virus collection time and high titer. The cell matrix is an ideal cell matrix which can be used for culturing the three viruses.

In cell instance segmentation of an original cell image obtained by microscopy, the original cell image is rotated by a rotation angle to yield a rotated cell image. A first machine-learning model for cell instance segmentation processes the original and rotated cell images such that effectively, horizontal and oblique boxes are used in bounding cells of the original cell image for enhancing cell-segmentation accuracy. A set of predictions is generated for each cell instance identified from the original and rotated cell images, yielding a plurality of sets of predictions for all cell instances. The plurality of sets of predictions is processed to remove any set having predicted space not simply connected, any unwanted set classified by a second machine-learning model as a poor indicator according to a certain performance criterion, and any redundant set. The plurality of sets of predictions then produces a segmentation map for the original cell image.

The invention discloses a method for large-scale production of engineered extracellular vesicles based on freeze-drying and hydration, which comprises the following steps: carrying out freeze dehydration treatment on cells, and then inducing the cells to generate target extracellular vesicles through hydration treatment. The method provided by the invention has good cross-cell type compatibility, is adaptive to suspension cells and adherent cells through a mild membrane reconstruction process, and has good biological safety. The invention further discloses the extracellular vesicles produced by the method and application of the extracellular vesicles.

The present disclosure relates to methods and kits for generating single cell barcodes and imparting them to the constituent molecules within a single cell. Additionally, methods to overlay sample barcode and spatial barcode information onto the single cell barcodes are also described. Generation of single cell barcodes is achieved by labeling the genomic DNA of a cell / nucleus with a small handful, preferably just a one or two cellular barcode probes (CBP) that can be amplified and propagated to label the constituent molecules within the cell. The disclosure finds utility in applications such as characterization of cellular heterogeneity, comprehensive profiling of tissue composition, characterization of adherent cells, discovery of new cell subtypes and functions of individual cells in the context of its microenvironment, and others.

To provide a cell proliferation system, proliferation method, control method, and culture medium bag capable of proliferating various adherent cells and suspension cells. [Solution] Cells are grown in a bioreactor and activated by an activator (e.g., a soluble activator complex). The nutrient supply and gas exchange functions of an automated closed cell proliferation system make it possible to seed at, for example, a low seeding density. By manipulating the parameters of the cell growth environment, cells are placed in specific locations within the bioreactor for efficient exchange of nutrients and gases. System parameters are adjusted to shear any cell colonies that may form during the proliferation phase. Metabolic concentrations are controlled to improve cell growth and viability. Cell retention within the bioreactor is controlled. In some embodiments, the cells include T cells. In other embodiments, the cells include a T cell subpopulation, for example, regulatory T cells (Tregs).

The utility model discloses a culture device for a cell migration experiment, which relates to the field of cell experiment equipment and comprises a culture dish and a cell culture plate with various hole site specifications such as 24 holes, 12 holes, 6 holes and the like, the culture device structurally comprises a bottom plate and a hole plate at the top of the bottom plate, and a plurality of culture hole sites are distributed at the top of the hole plate in a rectangular array manner. The culture hole site is used for adherent cell culture, the bottom of the culture hole site is provided with at least one membrane strip, the top of the membrane strip is provided with grid scale marks, and the bottom of the culture hole site and the bottom of the membrane strip are also provided with grid scale marks. The scratching step of a cell migration experiment can be simplified in the process of a cell culture experiment, the scratching device is convenient to use and can be used at any time when being taken, and the cell migration scratching experiment can be carried out for multiple times in one experiment.

The utility model discloses a differential adherent cell separation device which comprises a separation box body and an upper box cover, culture cavities and a one-way conducting assembly are arranged in the separation box body, the one-way conducting assembly is arranged between the culture cavities, the side face of the separation box body extends outwards to form a supporting part, the side edge of the supporting part protrudes upwards to form a baffle ring part, and the baffle ring part is connected with the upper box cover. A limiting slot is formed among the baffle ring part, the supporting part and the separating box body, and the four sides of the upper box cover extend downwards to form side ring parts. According to the utility model, the one-way conduction between the culture cavities is realized through the one-way conduction assembly, the communication state between the culture cavities can be controlled at a proper stage according to the actual condition of cell wall attachment, the cells are ensured to flow according to the expected direction and mode, and the accuracy and purity of cell separation are effectively improved; the upper box cover and the separation box body are tightly and stably connected, the upper box cover is prevented from loosening or displacing in the using process, and the overall sealing performance and stability of the device are guaranteed.

The invention discloses a multilayer adherent cell culture reactor and a use method. The culture reactor comprises a shell, a plurality of layers of cell culture plates are arranged in the shell in an array manner; and the liquid inlet is used as an inlet of a cell suspension. The liquid outlet is an outlet of the culture medium; the air holes are formed in the upper surface and the lower surface of the shell. And the quality control chamber is communicated with the inner cavity of the shell. The use method comprises the following steps: uniformly mixed cells and a culture medium enter the inner cavity of the shell through the liquid inlet for positive culture; after front culture is completed, the multi-layer adherent cell culture device is inverted; uniformly mixed cells and a culture medium enter the inner cavity of the shell through the liquid inlet for reverse culture. By arranging the multiple layers of cell culture plates with double surfaces capable of culturing, the cell culture area is greatly increased, the space utilization rate is improved, the use amount of a culture medium is reduced, the prepared quality control chamber is communicated with the inner cavity of the shell, and the cell growth state is conveniently, rapidly and accurately observed on the premise of not polluting the internal environment and not damaging the culture stability through isolation of the partition opening.

The invention belongs to the technical field of bioelectricity medicine and cell therapy, and particularly relates to a system for carrying out intracellular electric treatment based on ultrahigh-frequency ultrashort pulses and application of the system. The system comprises a high-voltage ultrashort pulse generation module, a control module and an adherent cell processing module, wherein the high-voltage ultrashort pulse generation module is respectively connected with the control module and the adherent cell processing module; the control module presets a plurality of pulse parameter combinations, the high-voltage ultrashort pulse generation module outputs a pulse electric field with preset parameters according to an instruction sent by the control module, and the pulse electric field acts on an organelle through the adherent cell processing module. High-frequency ultrashort pulses are adopted, the unique electric field distribution characteristic of the high-frequency ultrashort pulses can penetrate through cell membranes without causing permanent damage, and the high-frequency ultrashort pulses directly act on specific organelles such as mitochondria, endoplasmic reticulum and lysosome. And the system can flexibly induce four main programmed cell death modes, is suitable for different treatment requirements, and is beneficial to precise medical treatment.

The application relates to a biological cell culture device and a culture method, and relates to the technical field of cell culture. The device comprises a culture box main body with an open upper side. A placing mechanism and a pressing mechanism are arranged in the culture box main body. Before a cylinder is removed, the upper end of the cylinder is first sealed by a sealing plate. Therefore, after the culture dish is removed, external air cannot enter the culture box main body from the corresponding cylinder. When the subsequent culture dish enters, only the gas with the same volume as the cylinder enters the box, and the influence on the gas concentration in the culture box main body is not obvious. The gas concentration environment in the box is protected. Then, the adherent cells are separated from the inner wall to form a suspension liquid through centrifugation. The splashing of the culture liquid is avoided by the sealing plate. Then, the culture liquid is stirred at a low speed by a stirring mechanism. Since the adherent cells are separated from the inner wall in advance and move with the culture liquid, the damage to the cells in the stirring process is reduced, and the uniform distribution of the culture liquid is ensured.

The present invention discloses a novel uniform strain chamber for an adherent cell loading device, which relates to the field of cell mechanics. Cells are planted in the novel uniform strain chamber of the adherent cell loading device, and the four positioning holes of the chamber are placed on a fixed pile. The fixed pile is connected to the stator. The controller controls the stepper motor, and the rotational motion of the motor is converted into linear motion through a transmission screw and transmitted to the mover, thereby indirectly strain stimulating the cells in the chamber cavity, thereby realizing relevant mechanical biology research on the cells. The strain of the adherent cell chamber in the existing cell tensile loading device is uneven, and the experimental results have large errors. In the invention, the elastic basement membrane of the chamber is designed to have one side as a plane, i.e., the part where the planted cells are poured into the cell culture medium, and the other side as a curved surface, i.e., an optimized surface. The longitudinal cross-section is an M-shaped structure, and its thickness variation meets the uniformity requirement of the strain of the plane part of the basement membrane during the chamber loading process, thereby increasing the uniform area of ​​the adherent cells in the strain loading, and therefore has high use value.

The disclosed invention relates to a method and system 10 for releasing adherent cells 12 in a cell container 14 with a plurality of acoustic transducers 16a, 16b, 16c, 16h. The method comprises: gene

This invention provides a method for processing live single-cell extraction based on deformation trends, comprising the following steps: Step S1: Controlling a microfluidic chip to immerse itself in a culture medium containing adherent cells while maintaining a preset gap, controlling the injection flow rate of the injection channel and the aspiration flow rate of the aspiration channel to form a microjet acting on the cells; Step S2: Controlling an image processing module to receive and process cell morphology images on the culture substrate acquired by a microscopic imaging module, and calculating deformation parameters based on the comparison between the cell area in the current frame image and the initial cell area; Step S3: Comparing the deformation parameters with critical deformation parameters obtained through experimental calibration, and when the deformation parameters are greater than the critical deformation parameters, controlling a fluid drive module to reduce the injection flow rate and / or the aspiration flow rate; or, calculating a comprehensive damage risk index based on the deformation parameters, and when it is greater than 1, reducing the injection flow rate and / or the aspiration flow rate.

A substrate for culturing cells that comprises a modified prolamin having a net positive charge is provided. The modified prolamin may be a prolamin that has been modified through amidation or esterification, such as the addition of a methyl-ester group or an ethanolamine group. The modified prolamin is animal-free may be food-grade and / or may be free of cross-linkers. The modified prolamin may be in a coating that can be applied to a surface for culturing adherent cells, such as a microcarrier (e.g., beads) or well plates. Substrates coated with the modified prolamin are able to support cell expansion and can be used for food-grade applications such as cultured meat.

In optical microscopy including fluorescence microscopy, a biological sample (215) can be immersed in an aqueous liquid medium (210) during observation. Due to the refractive index mismatch between the liquid medium and a conventional glass bottom container holding the sample, image resolution is reduced when viewing the sample. To improve resolution, a sample container (100) is formed with a container body (110) having a bottom (120) with an aperture (125), a fluorinated ethylene propylene copolymer (FEP) film (150) attached to the bottom (120) to cover the aperture (125) to form a cavity-facing surface (155) on the film (150), and exposed to the liquid medium (210). Since the refractive index of FEP is close to that of water, image resolution is improved. Furthermore, the cavity-facing surface (155) is activated by plasma treatment to promote adhesion of the cavity-facing surface (155) to the biological sample (215) to enable cells in the biological sample (215) to proliferate in adherent cell culture and to immobilize the biological sample (215) during observation.

The disclosure is directed to a method for seed train expansion of adherent cells comprising culturing cells with a serum-supplemented growth medium in a N-2 vessel; removing the cells from the serum-supplemented medium; inoculating the cells from step into a serum-free growth medium in a N-1 vessel; culturing the cells in the N-1 vessel under suspension conditions; and inoculating a growth medium in a bioreactor with the suspension-cultured cells. In some aspects, the adherent cells are not suspension-adapted. In some aspects, the adherent cells are suspension-adapted. In some aspects, the adherent cells produced by the seed train expansion method are used to produce viral vectors. In some aspects, the viral vectors are AAV vectors.

The current invention relates to the field of in vitro culturing of T cells, more in particular of in vitro production of unconventional T cells expressing specific markers. Indeed, the present invention discloses a method to produce said unconventional T cells comprising the generation of hybrid spheroids out of combining stem cells and adherent cells and further culturing said spheroids at the air liquid interface. The latter T cells can be used to produce CAR-T cells.

Disclosed is a system for automating passaging of anchorage dependent or adherent mammalian cells, having a fluid distribution cartridge where fluid channels each has an associated pneumatically controlled valve, and the valves can be selectively opened and closed to allow fluid which is pumped into channels in the cartridge to be selectively passed or interrupted. The channels are connected with a series of vessels each held in a housing structure, and a first frame rests on a surface and a second frame is attached with the first frame such that the second frame can tilt on a first axis with respect to the first frame, and the housing is attached to the second frame such that the housing can tilt on a second axis with respect to the second frame, and wherein the second axis is orthogonal to the first axis. A digital system that controls selective opening and closing of the valves by controlling the pressurized air provided to the valves, and also controls the movement of the frames and housing under feedback from the leveling system.

Methods comprise exposing a culture of human pluripotent stem cells adherent on a first substrate comprising a first laminin to a differentiation medium for a first time period of 15 days to 50 days to obtain a first population of adherent cells comprising RPE cells and / or progenitors thereof; at the end of the first time period, dissociating the first population from the first substrate; replating the dissociated first population of cells on a second substrate comprising a second laminin; and culturing the replated first population of cells on the second substrate for a second time period to obtain an expanded and matured second population of cells comprising the RPE cells. The first and second laminins are independently selected from LN-521, LN-511, LN-111 and LN-121, and are an intact protein or protein fragment. Also included are RPE cells and materials and compositions utilizing such RPE cells for various treatments.

The invention relates to the technical field of suspension cell domestication methods, in particular to a fish cell suspension domestication method and application. The serum-free culture medium used in fish cell suspension domestication provided by the invention contains one or more of amino acid, vitamin, metal salt, lipid, trace elements, a buffer agent or an acid-base indicator. According to the domestication method provided by the invention, fish adherent cells are directly cultured by adopting a suspension culture medium under a suspension condition until the cells realize forward proliferation, single-cell full-suspension fish cells are obtained, and serum-free domestication is performed while suspension domestication is performed, so that the fish suspension cells which can be cultured under a serum-free condition are obtained. The method successfully realizes single-cell suspension domestication of fish cells, the cell growth state is stable, the monodispersity is good, and the method has high-density proliferation potential, is suitable for single-cell full-suspension culture in a serum-free culture medium, and can be used for large-scale production in a bioreactor.

The invention belongs to the technical field of life science analysis instrument equipment and clinical examination and detection equipment, and particularly relates to a cell sorting system for in-situ reverse killing of adherent cells, which comprises a sample injector, a micro-fluidic chip, a light path system, a killing signal generator and a computer control unit, the micro-fluidic chip comprises a flowing layer, an upper-layer electrode and a combined electrode layer which are sequentially arranged from top to bottom, the combined electrode layer at least comprises a photosensitive electrode, and the killing signal generator is used for loading killing electric signals to the upper-layer electrode and the combined electrode layer to form a high-intensity electric field to kill cells to be killed. In conclusion, aiming at the adherent cells, the invention comprehensively provides a single cell precise reverse killing scheme based on light-electricity cooperation, and the killing accuracy and efficiency of the to-be-killed cells can be effectively improved.

To provide compositions and methods that facilitate transfection of cells.SOLUTION: Methods and compositions containing ethanol and an isotonic salt solution are used for delivery of compounds and compositions to non-adherent cells, for example, T cells.SELECTED DRAWING: Figure 10

The invention relates to biological cell culture equipment and a culture method, and relates to the technical field of cell culture.The biological cell culture equipment comprises a culture box body with an opening in the upper side, a placing mechanism and a pressing mechanism are arranged in the culture box body, before a culture dish is moved out of a cylinder, the upper end of the cylinder is sealed through a sealing plate, and therefore after the culture dish is moved out subsequently, the pressing mechanism is pressed; external air cannot enter the incubator main body from the corresponding cylinder, only gas with the same internal volume of the cylinder enters the incubator when a subsequent culture dish enters the incubator main body, the gas concentration in the incubator main body is not obviously influenced, the gas concentration environment in the incubator is protected, and then adherent cells are separated from the inner wall to form suspension liquid in a centrifugal mode. The sealing plate is used for avoiding splashing of the culture solution, then the stirring mechanism is used for stirring the culture solution at a low speed, adherent cells are separated from the inner wall in advance and move along with the culture solution, damage to the cells in the stirring process is reduced, and it is guaranteed that substances of the culture solution are evenly distributed.

The present application relates to the technical field of biology, and particularly discloses a method for culturing and obtaining foregut endoderm cell spheroids and application thereof. The method for culturing and obtaining foregut endoderm cell spheroids of the present application obtains foregut endoderm cell spheroids by inducing differentiation of stem cells on a culture surface with a PDA-matrigel coating. The culture method of the present application can ensure the differentiation efficiency of DE and AFE adherent cells and spheroids, promote the generation of AFE cell spheroids and the separation from the adherent cell layer, reduce the edge rolling of AFE adherent cells, prolong the production time of AFE cell spheroids, significantly improve the yield of AFE cell spheroids, and provide a basis for subsequent differentiation and culture of lung organoids.

This invention relates to the field of cell engineering technology, and more particularly to a method for preparing immunomodulatory stem cells for treating graft-versus-host disease (GVHD). The invention provides a method for preparing immunomodulatory stem cells for treating GVHD, comprising the following steps: (1) washing human fat aspirate and digesting it with collagenase to obtain monocytes; (2) inoculating the monocytes obtained in step (1) into differentiation medium and culturing for 2–3 days, then continuing to culture the adherent cells for 18–22 days to obtain mesenchymal stem cells; (3) inoculating the mesenchymal stem cells obtained in step (2) into induction medium for induction culture, and after screening, obtaining immunomodulatory stem cells. The preparation method provided by this invention solves the technical problems of unstable sources, functional decline, and poor therapeutic effects of immunomodulatory stem cells in the prior art, providing a new, efficient, safe, and standardized solution for the treatment of GVHD.

Provided herein are altered cell lines comprising a suspension cell line adapted from an adherent cell line having a different expression profile from a corresponding non-altered adherent cell line, methods for generating altered cell lines, and methods of characterizing altered expression profiles for a gene, a transcript, or a protein in an altered cell line.