217 results about "Adherent cell" patented technology

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.

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 the field of biological medicine, in particular to a water-soluble alginate microcarrier as well as a preparation method and application thereof. The alginate microcarrier comprises a carrier core structure and a shell surface modifier, the micro-carrier core layer is formed by carrying out plasma crosslinking on water-soluble alginate, ethylenediamine tetraacetic acid salt, nitrilotriacetic acid salt, diethylene triamine pentaacetic acid and water-soluble divalent metal salt, and light-sensitive alginate and a photoinitiator can also be added, so that the micro-carrier core layer has two crosslinking networks; the shell layer is composed of one of gelatin, collagen, fibrinogen, albumin and fibrin, or the gelatin, the collagen, the fibrinogen, the albumin and the fibrin and other biocompatible polymers; the obtained microcarrier has the characteristics of high transparency and rapid degradation, and adherent cells can be amplified in vitro; the microcarrier obtained in the invention can be used for mesenchymal stem cell culture, and can be rapidly degraded under a mild cell condition to harvest cells.

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 embodiment of the invention is applicable to the technical field of biomedicine and image processing, and provides a cell confluence degree calculation method, a cell confluence degree calculation device and computer equipment. According to the method, an adherent cell original image is processed by using opening operation, closing operation and the like in a morphological image processing algorithm, and contrast enhancement is combined, so that the confluence degree of cells is obtained. The original image is optimized by brightness equalization means such as gamma correction, the pixel gradient change of the stem cell region image can be enhanced, and the characteristics of the stem cell image are highlighted. And then, locking a cell region by using an edge detection algorithm, and obtaining a complete cell region through algorithms such as image connected domain calculation, flooding filling and optimized region growth. On the basis, stem cell image cell region segmentation and cell confluence degree calculation can be quickly realized based on a traditional algorithm. By applying the calculation method of the cell confluence degree provided by the embodiment of the invention, the calculation of the cell confluence degree can be quickly and accurately realized.

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.

To provide methods of producing and purifying AAV viral vectors having low empty capsids, low host cell proteins, and / or low contaminating DNA, while retaining high potency; and to provide a pharmaceutical composition containing the vector.SOLUTION: A method of manufacturing an AAV viral vector comprises: a. culturing adherent cells; b. transfecting the adherent cells with one or more plasmids to enable production of the AAV viral vector; c. lysing the adherent cells to isolate the AAV viral vector; d. acidifying and clarifying the cell lysate of c; e. purifying the product of d using cation exchange chromatography; f. filtering the product of e using tangential flow filtration; g. ultracentrifuging the product of f in a cesium chloride buffer; h. collecting the AAV viral vector from the product of g; and i. formulating and filling the AAV viral vector collected in h. to produce a final pharmaceutical product.SELECTED DRAWING: None

Provided are establishment and suspension acclimation of a Crandell Reese Feline Kidney (CRFK) adherent cell line, and its application. The CRFK cell line, named CRFK-BLA, is deposited at the China General Microbiological Culture Collection Center (CGMCC), with the address being Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, the deposit number being CGMCC NO: 45703; the CRFK cell line was classified and named CRFK cells, and was deposited on Aug. 17, 2023. A serum-free complete suspension culture type CRFK cell line obtained by acclimation on the basis of CRFK-BLA is named CRFK-BLS, and is deposited at the China General Microbiological Culture Collection Center (CGMCC), with the address being Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, the deposit number being CGMCC NO: 45704; the CRFK cell line was classified and named CRFK suspension cells.

The invention discloses a method for realizing in-vitro amplification of adherent cells by utilizing a stirring type bioreactor matched with a three-dimensional porous hydrogel carrier. The method comprises the following steps: sterilizing a hydrogel carrier and a stirred bioreactor, adding a cell culture medium, ventilating, inoculating cells, starting the stirred bioreactor for culturing, observing the color change of a culture solution in the culture process, and changing the solution when the culture solution turns yellow, thereby ensuring the nutrition required by cell culture. The hydrogel with high biocompatibility is used as a carrier, a porous three-dimensional growth environment is provided, cells can be attached to the interior of the hydrogel carrier, the cell growth space is sufficient, the cells can be cultured for more generations, the number and density are greatly increased, the amplification multiple can reach 10 times or above, the production efficiency is improved, and the production cost is reduced.

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 invention discloses a construction method of an endothelial progenitor cell model for pulmonary arterial hypertension research, which comprises the following steps: (1) enriching and separating mononuclear cells centrally existing in CD34 cells in peripheral blood of a pulmonary arterial hypertension patient by adopting Histopaqui-10771, then culturing and resuspending the mononuclear cells in an EGM-2MV culture medium containing growth factors, then spreading in a culture plate coated with fibronectin for culturing; (2) 2 days after planking, sucking away non-adherent cells; and cleaning the adherent cells, adding the EGM-2MV culture medium containing the growth factors again for continuous culture, changing the culture medium and adding fibronectin at regular time during culture, inducing directional differentiation of the endothelial progenitor cells, and culturing for 2 weeks to start to appear the endothelial cells. The injury state of the endothelial cells in the disease is represented by the endothelial cells differentiated from the endothelial progenitor cells of the pulmonary hypertension patient, the research on the pulmonary hypertension cells and molecular mechanisms is facilitated, and the method is used for screening pulmonary hypertension treatment drugs and exploring potential pulmonary hypertension treatment targets.

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 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 polymer; a cultivation method for adherent cells which comprises two-dimensional culturing of adherent cells after preservation via said preservation method; and a cell preservation liquid which includes a high polymer that is suitable for use in these methods.

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 utility model relates to a cell culture device capable of conveniently changing liquid, which comprises a culture plate, a liquid collecting plate and a liquid changing plate which are respectively provided with a plurality of culture bins, a plurality of liquid collecting holes and a plurality of liquid changing holes, and has a liquid collecting state, a liquid discharging state and a liquid changing state. In the liquid collecting state, the culture plates are stacked on the liquid collecting plates, so that each culture bin is contained in the corresponding liquid collecting hole, and a protruding part arranged at the bottom of each liquid collecting hole is embedded into and blocks a through opening formed in the bottom wall of the corresponding culture bin. The culture plate is lifted from the liquid collecting plate so that the protruding parts of the liquid collecting holes are drawn out from the through openings; in the liquid changing state, the culture plates after liquid discharging are stacked on the liquid changing plate, so that each culture bin is contained in the corresponding liquid changing hole, and the new culture liquid in the liquid changing hole enters the culture bin through the through opening of the culture bin. According to the cell culture device, rapid liquid change can be realized without a pipettor, and adverse effects on adherent cells caused by liquid change of the pipettor are avoided.

The invention discloses a cellular exosome-coated hexagonal boron nitride / polyvinylpyrrolidone nano-particle as well as a preparation method and application thereof, relates to the technical field of medicines, and aims to solve the problem that in the prior art, in BNCT tumor treatment, the BNCT treatment effect is poor. In order to solve the problems of poor targeting and biocompatibility of a boron carrying agent, insufficient in-vivo stability and tumor accumulation capacity of nanoparticles and the like, the boron carrying agent comprises hexagonal boron nitride-polyvinylpyrrolidone nanoparticles and cell exosomes wrapping the peripheries of the hexagonal boron nitride-polyvinylpyrrolidone nanoparticles, and cells are adherent cells. The targeting property and biocompatibility of the boron carrying agent and the in-vivo stability and tumor accumulation capacity of nanoparticles can be effectively enhanced, and the BNCT treatment effect of glioma is improved.

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.