161 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.

To provide a cell preservation liquid and a cell preservation method which enable long-term refrigerated preservation of adherent cells while maintaining adherence capability to a culture substrate.SOLUTION: The present invention relates to: a preservation method for adherent cells which comprises refrigerated preservation of adherent cells in a non-frozen state in a cell preservation liquid that contains a high-molecular-weight; a cultivation method for adherent cells which comprises two-dimensional culturing of adherent cells after preservation via the preservation method; and a cell preservation liquid which includes a high-molecular-weight that is suitable for use in these methods.SELECTED DRAWING: None

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.

A cell culture matrix for culturing cells in a fixed bed reactor is provided. The cell culture matrix includes a first substrate material having a regular and orderly array of openings passing through the layer, the openings being separated by a substrate material having a physical structure that is substantially regular and uniform and configured such that cells grow thereon. The physical structure and array of the openings are configured such that when the first substrate material is rolled into a roll-shaped substrate matrix including a roll-shaped layer of the first substrate material, the variation in the packing density of the roll-shaped layer is less than about 20 times, less than about 10 times, less than about 5 times, or less than about 2 times throughout the roll-shaped substrate matrix.

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.

A method of generating MSC-derived exosome populations may include collecting MSC containing material from living tissue, separating desired mononuclear cells from granulocytes, culturing to multiply the cells, separation of desired cells for further multiplication by washing non-adherent cells and culturing adherent cells, repeating as necessary to obtain a suitably pure population of MSCs, culturing the MSCs in culture media containing negative / healing active cytokines interleukin-4 (IL-4) and interleukin-10 (IL-10) and multifunctional cytokine TGF-ß, and isolating the MSC-derived exosome populations. Diverse MSC-derived exosome populations may be generated by altering the cytokine composition of the culture media. The MSC-derived exosome populations may be screened for effectiveness in treatment of Long Covid using in vitro, in vivo, and pre-clinical testing utilizing model organisms. The exosomes may be administered nasally. Successful MSC-derived exosome populations may be further subjected to patient trials to establish efficacy in treatment of Long Covid via nasal administration of the MSC-derived exosome populations to human subjects. Similar methodologies may be employed to establish efficacy of the MSC-derived exosome populations for treatment of other diseases and conditions related to the central nervous system, spinal cord injury, or neurological diseases, such as Alzheimer disease.

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.

Provided in the present invention is a cell freezing medium for clinical use. In particular, the cell freezing medium of the present invention comprises the following components: (1) human albumin; (2) cryoprotectant: the cryoprotectant comprises a combination of one or more of dimethyl sulfoxide, glycerol, and ethylene glycol; (3) a saline buffer; wherein the salt buffer is a solution containing Na+, K+, Mg+, Cl−, and CH3COO− ions; (4) a vitamin; and (5) an amino acid, wherein the human albumin concentration is 1%-20% (w / v). The cell, after long-term cryopreservation with the freezing medium of the present invention, has a high viability, and the cellular efficiency maintains a high uniformity. The grade of purity of the freezing medium of the present invention is the pharmaceutical grade or USP grade; and the freezing medium is safe and reliable for clinical use, and can be used or conventional adherent and suspension cells.

Problem: To provide a cell culture substrate, a cell culture vessel, and a cell detachment method for adherent cells, which enable easier detachment of adherent cells compared to a case where trypsin is used. Solution: A cell culture substrate for coating a cell culture vessel, the substrate containing, as an active ingredient, one or more proteins of a silk protein, a silk protein hydrolysate, fibroin, and a fibroin hydrolysate.

A biological tissue formation device capable of efficiently performing cell-to-cell interaction and liquid component exchange between cell layers of a formed biological tissue is provided. A biological tissue formation device (1) is provided for forming a biological tissue having multiple cell layers composed of adherent cells (30, 31), comprising: a culture membrane (10) having culture areas for adherent cells (30, 31) on both sides, wherein the cultured adherent cells (30, 31) are placed between the multiple cell layers, and multiple flow paths (110, 130) are divided by the culture membrane (10); the culture membrane (10) is composed of a soluble material (101).

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 invention relates to the technical field of cell engineering, in particular to a preparation method of immune regulation type stem cells for treating graft versus host diseases. The invention provides a preparation method of immune regulation type stem cells for treating graft versus host diseases, which comprises the following steps: (1) cleaning a human fat aspirator, and digesting with collagenase to obtain mononuclear cells; (2) inoculating the mononuclear cells obtained in the step (1) into a differential culture medium, culturing for 2-3 days, and further culturing the adherent cells for 18-22 days to obtain mesenchymal stem cells; and (3) inoculating the mesenchymal stem cells obtained in the step (2) into an induction culture medium for induction culture, and screening to obtain the immune regulation type stem cells. According to the preparation method provided by the invention, the technical problems of unstable source, function attenuation and poor treatment effect of the immune regulation type stem cells in the prior art are solved, and an efficient, safe and standardized new scheme is provided for treatment of GVHD.

The utility model discloses an adherent cell culture medium, which belongs to the technical field of culture media for adherent cells and comprises a shell, a partition plate arranged in the shell, four cavities arranged between the partition plate and the shell, a culture dish arranged in each cavity, and a solid culture medium arranged at the bottom of the inner side of each culture dish. A pop-up assembly is arranged between the culture dish and the shell, the pop-up assembly is used for popping up the culture dish, the pop-up assembly comprises a threaded rod, the threaded rod is arranged on the lower side of the culture dish, the lower side of the shell is rotationally connected with a nut, the nut is in threaded connection with the threaded rod, and a plurality of springs are arranged between the culture dish and the shell; and an adjustable shading component is arranged on the upper side of the shell and is used for performing different shading adjustments on the culture dishes in the cavity, so that the problem that a plurality of groups of adherent cells are inconvenient to culture and observe by using the same culture medium is solved.

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.

The present invention provides: a method for culturing adherent cells in a culture vessel using a microcarrier covered with a polymer showing a lower critical solution temperature, said method being characterized by comprising the following steps (1)-(4): (1) culturing adherent cells in a culture solution having a temperature equal to or higher than the lower critical solution temperature in a culture medium having a temperature equal to or higher than the lower critical solution temperature; a step for culturing the cells on the surface of the microcarrier; (2) a step in which, after step (1), the culture solution is cooled to a temperature equal to or lower than the lower critical solution temperature; (3) a step in which, after step (2), the culture solution in the culture vessel is stirred to peel the cells from the surface of the microcarrier; and (4) a step for recovering the cells peeled off from the surface of the microcarrier after step (3), the temperature-responsive microbeads comprising a polymer coating film and a carrier, the polymer coating film comprising a temperature-responsive polymer having a lower critical solution temperature at 0-50 DEG C, and the carrier comprising a polymer having a lower critical solution temperature at 0-50 DEG C, and the polymer coating film comprising a polymer having a lower critical solution temperature at 0-50 DEG C and a polymer having a lower critical solution temperature at 0-50 DEG C and a polymer having a lower critical solution temperature at 0-50 DEG C. The surface of the carrier has positive charges, and the film thickness of the temperature-responsive polymer is 10-1000 nm.