59 results about "Cell aggregation" patented technology

The present invention relates to a microfluidic device (1) for forming a cell aggregate comprising at least one first cell (C1) and one second cell (C2), and for individually processing selected cells of the cell aggregate: - Microfluidic channel (10); - At least one main inlet (11) for fluids containing a first cell, a second cell, and a third cell, respectively, located in the first portion (101) of the microfluidic channel; - An outlet (12) located in the second portion (102) of the microfluidic channel for controlling the flow rate of fluid within the microfluidic channel; - A first auxiliary inlet (131) for at least one first auxiliary fluid, located in the first portion (101) of the microfluidic channel upstream or downstream of the main inlet (11); - At least one cell trapping section (14) positioned between the first and second parts within the microfluidic channel; - At least one first valve to control the flow rate of the first auxiliary fluid, causing the fluid containing the first cells and the second cells to flow at a predetermined height within the microfluidic channel, thereby guiding the first cells and the second cells to the first and second capture units, respectively. Equipped with, The microfluidic device relates to a microfluidic device in which each cell capture section (14) comprises at least one first capture section (141) and one second capture section (142), each first and second capture section being sized to accommodate a first or second cell, the first and second capture sections being adjacent to each other in a direction perpendicular to the bottom (100) of the microfluidic channel, and forming a cell aggregate containing the captured first and second cells, each cell being at a different height relative to the bottom of the microfluidic channel.

This invention provides a method for propagating pluripotent stem cells (PSCs) by suspension culture in a bioreactor. [Solution] The method comprises the following steps: (i) adding a ROCK inhibitor (ROCKi) to pluripotent stem cells cultured in suspension in a bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding a sufficient excess volume of culture medium to reduce the concentration of the cell dissociation agent to a concentration at which cell aggregates can be re-formed; and (iv) culturing the mixture obtained in step (iii) under appropriate conditions that allow for the proliferation of PSCs.

In a bioprinting system and method, a bioprinter may include one or more reservoirs, a printhead, and a print surface on which the bioprinting is dispensed. An optical system may collect imaging data of a plurality of cells within a cross-linkable material during and / or after passage of the plurality of cells through the bioprinting system. A computing system may generate, via a machine learning algorithm executing on the image data, a plurality of segmented regions in the image data, wherein each segmented region in the plurality of segmented region defines a boundary around a different set of cell aggregates in the plurality of cells, identify a first segmented region in the plurality of segmented regions that defines a boundary around at least a visible portion of a first cell aggregate, wherein the first cell aggregate includes an occluded portion, determine, based on an estimated shape of the first cell aggregate, an extrapolated boundary around at least some of the visible portion of the first cell aggregate and at least some of the occluded portion of the first cell aggregate, compute a quantity of cells within the first cell aggregate based on an area surrounded by the extrapolated boundary, and determine one or more parameters associated with the plurality of cells, the material flow, or the bioprinting system based on the quantity of cells.

The present disclosure relates to a method of generating pancreatic lineage cells, e.g. islet-like cell aggregates comprising pancreatic beta cells, comprising the steps of providing a single cell suspension of a population of endocrine progenitor cells (EP), allowing the EP cells in the single cell suspension to form 3D structures, and culturing the cells under conditions allowing differentiation into pancreatic mono-hormonal beta cells. The present disclosure also relates to islet-like cell aggregates obtainable by said method and their medical use.

The present invention relates to novel cell culture and transfection methods using a wildtype Caf1 ("Caf1-WT") polymer or a variant thereof. Use of a Caf1-WT polymer or a variant thereof to increase cell viability, cell proliferation, cell migration and / or cell clustering; promote the formation of a 3D cell structure; and / or promote suspension cell culture of cells that are normally adherent are also provided herein.

The invention provides a whole-brain organoid based on human induced pluripotent stem cells as well as a construction method and application thereof, and belongs to the technical field of organoid construction. The method comprises the following steps: culturing human induced pluripotent stem cells in a complete culture medium to obtain a cell aggregate; culturing in an embryoid body forming culture medium to obtain a spherical embryoid body; culturing in a nerve induction culture medium tissue to form a nerve epithelium structure; embedding the single embryoid body with the nerve epithelium structure in a Matrigel gel drop, culturing in a nerve expansion culture medium to form a nerve epithelium-like structure, and replacing a mature culture medium for culturing to obtain the whole-brain organ. According to the method, by dynamically and accurately regulating and controlling culture parameters, the control ability of a cell differentiation area is improved, and interference of non-target cell types is reduced, so that the efficiency and consistency of embryoid formation and neural differentiation are directly improved, and finally, the obtained mature organoid has a more complete epithelial structure and cell polarity.

The application discloses a dispersion mixed solution system capable of reducing cell aggregation in a domestication process and a preparation method thereof. The system comprises 50x concentrated A liquid (100-250 g / L trypsin+12.5 g / L EDTA) and 100x concentrated B liquid (100 g / L P188+10 g / L DS), and realizes long-term stable storage and rapid preparation in the form of concentrated stock solution. When used, the concentrated stock solution is diluted and mixed in proportion, and through the synergistic effect of mild cell lysis and weakening of cell aggregation force, the cell aggregation in the domestication process is efficiently dispersed, and the cell activity and function are maintained. The preparation method is simple, the storage period is long, the dispersion effect is good, and the cell damage is small, and the application provides an effective technical means for cell domestication.

This invention provides a method for efficiently manufacturing three-dimensional structures using food ink (or cell aggregate ink) containing impurities such as granular or fibrous fragments, and also provides food three-dimensional structures or cell aggregate structures manufactured by these methods. [Solution] Multiple frames prepared for molding can be fixed to each other by inserting fixing rods or the like into common positioning holes provided in the frames. Food ink is dispensed into each filling area within the frames, spread and filled by a spreading means, and the surface is smoothed. Then the upper frames are stacked sequentially. These operations are repeated to form a composite. After the composite is fixed, the frames can be removed to obtain a three-dimensional structure (M). M can be used as is, decorated with a food 3D printer, or heated in a microwave or steamed to make it suitable for eating.

A gellable material for encapsulating a cellular aggregate in a well plate is provided. The cellular aggregate, before or after encapsulation, can be organoid or a spheroid, and typically, the cellular aggregate, once encapsulated, has substantially no movement during media exchange, handling of the well plate or tilting of the well plate. Methods for producing encapsulated cellular aggregates, and uses thereof are also provided.

The present invention relates to the field of biotechnology, and more particularly to methods for characterizing, typing, and identifying cells and their applications. The physical information of cells includes the mechanical force and / or stiffness of cells obtained in at least one of the following cases: interactions between cells and / or multicellular aggregates, cells and / or multicellular aggregates at different growth stages, different regions within multicellular aggregates, the effects of substances on cells and / or multicellular aggregates, and the effects of other physical, biological, or chemical factors on cells and / or multicellular aggregates. The present invention performs typing and identification of cells and / or multicellular aggregates by the above-described characterization method. The present invention can characterize the real-time and continuous state of cells and / or multicellular aggregates by the physical information of cells, and can identify each type and state of cells and / or multicellular aggregates in a short time, at low cost, and with high throughput, with an accuracy of 98% or more. The present invention is further limited to being realized by a characterization system, and the above effects can be achieved.

Disclosed herein are methods of analyzing a tissue sample using quantitative phase contrast microscopy, as well as corresponding microfluidic systems and corresponding devices. The methods include providing the tissue sample in a sample volume of a microfluidic system, wherein the tissue sample includes a plurality of biological cells forming a continuous tissue material. At least a portion of the tissue sample is lysed into single cells and / or cell aggregates in a carrier fluid in the sample volume. A carrier fluid flow is generated from the sample volume to a measurement volume of the microfluidic system, and a first phase shift image of the single cells and / or cell aggregates in the measurement volume is acquired with a quantitative phase contrast microscope.

The method for producing a cell aggregate including glial progenitor cells according to the present invention comprises:(1) a step of subjecting pluripotent stem cells to suspension culture in an embryoid-body-forming culture medium containing one or more SMAD signaling inhibitors and one or more Wnt signaling activators in the absence of feeder cells for 5 days to 10 days, to form a cell aggregate;(2) a step of subjecting the cell aggregate obtained in (1) to suspension culture in an embryoid-body-forming culture medium containing retinoic acid;(3) a step of subjecting the cell aggregate obtained in (2) to suspension culture in an embryoid-body-forming culture medium or neuron-and-glia-proliferating culture medium containing retinoic acid and one or more SHH signaling activators; and(4) a step of subjecting the cell aggregate obtained in (3) to suspension culture in a neuron-and-glia-proliferating culture medium containing no retinoic acid and one or more SHH signaling activators.

In a method for implantation of a physically stabilized aggregate of living cells or tissue fragment is injected into a channel provided in soft tissue filled with an aqueous gel. Also discloses are methods of stabilizing such aggregates and fragments and of forming such channel in soft tissue as well as means for carrying out the methods.

The present invention relates to a device (1; 10; 15) for the storage and / or transport of cells, in particular cell aggregates, comprising at least one chamber (2) having a base (5; 11) and a top (6; 12), the top (6; 12) being located opposite the base (5; 11), the base (5; 11) being formed by a substrate comprising at least one recess (7) for collecting a fluid, wherein the distance (8) between the base (5; 11) and the top (6; 12) of at least one chamber (2) is less than the width (9) of at least one recess (7), and the width (9) of at least one recess (7) is the smallest line segment between two opposite points passing through the center of the base region of at least one recess (7).

Provided is a culture system capable of generating a sufficiently grown cell aggregate. A culture system that forms a cell aggregate by liquid culture includes: a culture portion that contains a liquid and has an outlet through which the liquid is discharged and an inlet through which the liquid is introduced; a path portion that allows the liquid to flow between the outlet and the inlet; a monitoring portion capable of monitoring a component in the liquid and / or properties of the liquid in at least one of the culture portion and the path portion; an adjustment portion capable of adjusting the component in the liquid and / or the properties of the liquid in at least one of the culture portion and the path portion; and a control portion capable of controlling the adjustment portion on the basis of a monitoring result of the monitoring portion.

The present disclosure relates to a method for the generation of cells of the pancreatic lineage, for example pancreatic islet-like cell aggregates comprising pancreatic β-cells, which method comprises the steps of providing a single cell suspension of a population of endocrine progenitor (EP) cells, allowing said EP cells in single cell suspension to form 3D structures and culturing said cells under conditions permissive of differentiation into pancreatic monohormonal β-cells. The present disclosure also relates to pancreatic islet-like cell aggregates obtainable by said method as well as to medical uses thereof.

Disclosed is a highly versatile method for preserving and transporting a cell aggregate capable of preserving and transporting a cell aggregate while maintaining the form of the cell aggregate. The method of preserving the cell aggregate includes a step of filling a filling vessel with a cell aggregate suspension such that the ratio of the volume of the cell aggregate suspension to the capacity of the filling vessel is 60% or higher, and a step of preserving the filling vessel in a non-frozen state for one hour or longer.

A method of monitoring one or more cell aggregates, includes providing a flow path in which the one or more cell aggregates are in a medium and the flow path being configured to pass through a collective sensing zone of a set of electrodes, obtaining impedance-related signals corresponding to each of the medium and one of the one or more cell aggregates in the medium, and determining one or more electrical signatures for a cell aggregate, in which the one or more electrical signatures are based on impedance-related signals obtained from the set of electrodes. The method is one of dynamic testing at single-particle resolution. The electrical signatures may be an opacity and / or electrical size of the one or more cell aggregates, or electrical impedance spectroscopy-based electrical signatures. It is also to provide a microfluidic chip having a channel and electrodes for obtaining impedance-related signals.

Agglomeration and deformation that occur during culture of a three-dimensional cell aggregate are suppressed to create a three-dimensional cell aggregate having a linear shape. A method for producing artificial cartilage tissue, the method comprising: agglomerating cartilage progenitor cells to form cartilage progenitor spheroids; seeding the cartilage progenitor spheroids along grooves in a membrane having the grooves; culturing the cartilage progenitor spheroids in a state in which a culture medium is supplied from the obverse side and the reverse side of the membrane to fuse the spheroids together; and then maturing the spheroids into cartilage-like tissue and / or long bone progenitor cartilage organoids.

To provide a method for producing a cell aggregate having a hair regeneration ability, and a method related thereto.SOLUTION: A method for producing a cell aggregate comprises: plating epithelial cells and mesenchymal cells; and co-culturing the epithelial cells and the mesenchymal cells to form a cell aggregate having a hair regeneration ability. The co-culturing includes a matrix process for keeping the epithelial cells and the mesenchymal cells in a culture solution having dispersed therein type-I collagen or fibronectin.SELECTED DRAWING: None

This invention relates to suspended cells, specifically to an anti-suspension cell clumping complex and its preparation method, and its application. The anti-suspension cell clumping complex is characterized by comprising sodium dextran sulfate, sodium heparin, and non-essential amino acids. The complex of this invention can reduce the cell clumping rate after 4 days of large-scale suspended cell culture (7.18%-10.27%). The optimal formulation shows a clumping rate as low as 7.18% (≤10%), significantly superior to sodium dextran sulfate (DSS), sodium heparin, non-essential amino acid groups, and commercially available anti-clumping agents. It effectively solves the problem of cell aggregation and clumping in large-scale, high-density culture of cells adapted to suspension.

To provide a method for freezing a cell aggregate containing nervous system cells.SOLUTION: A method for freezing a cell aggregate containing neural cells having a three-dimensional structure, the method comprising the steps of: (1) contacting the cell aggregate containing the neural cells having a three-dimensional structure with a preservation solution at 0°C or higher and 30°C or lower before freezing; A step of preparing a cell aggregate immersed in a preservation solution, and (2) a step of cooling and freezing the cell aggregate immersed in the preservation solution obtained in step (1) at least from a temperature about 5 °C higher than the freezing point of the preservation solution to a temperature about 5 °C lower than the freezing point at an average temperature decrease rate of 2 to 7 °C / min.SELECTED DRAWING: None

Provided are: a cell aggregate containing adhesive cells and an extracellular matrix, and also containing survival cells at a depth of 30 [mu] m or more from the surface of the aggregate; disclosed is a method for producing a cell aggregate, which comprises supporting adhesive cells on a molded body that is formed from fibers containing an ethylene-vinyl alcohol copolymer and has a non-woven fiber structure, and culturing the cells while stirring.

Provided herein are, inter alia, genetically engineered human pluripotent stem cells, beta cells derived from genetically engineered human pluripotent stem cells, compositions, and cell aggregates, and methods of treating diabetes. The human pluripotent stem cells and beta cells provided herein including embodiments thereof may be genetically modified to express exogenous genes and / or inactivate endogenous genes.

The tissue analysis system and the co-culture casting system comprise at least a porous co-culture casting plate and a muscular tissue casting plate. The use method comprises the following steps: inoculating a plurality of neuron culture zones of a porous co-culture casting plate with a living neuron material, inoculating a plurality of muscle culture zones of a muscle tissue casting plate with a living muscle material, and culturing a plurality of co-culture tissues in the porous co-culture casting plate, these co-cultured tissues comprise a muscle cell culture cultured from a living muscle material and in situ neurospheres matured from the living neuron material in cell aggregation characteristics, and constricting the innervated portion of the living muscle material in each co-cultured tissue by stimulating the living neuron material.

The invention discloses a dispersion liquid for serum-free suspension domestication of CRFK cells and a use method of the dispersion liquid. The dispersion liquid is composed of a liquid A and a liquid B, the liquid A is an enzyme active component (containing 0.1%-0.2% of trypsin), and the liquid B is a protective component (containing 1%-3% of a serum substitute and 0.01%-0.05% of Prusich F-68). The core of the invention is a dynamic proportion adjustment application method, wherein the ratio of the solution A to the solution B is 2.5: 1 in the initial stage of domestication (1-3 generations); the proportion in the middle stage of domestication (the fourth generation to the fifth generation) is reduced to 1.5: 1; at the later domestication stage (after the sixth generation), the proportion is reduced to 1: 1, and meanwhile, serum-free TransVA-03 is used as a culture medium for culture under the shaking table conditions of 36.5 DEG C, 7% CO2 and 110r / min. The method solves the problem of agglomeration in the serum-free suspension domestication process of the CRFK cells, the viability of the domesticated cells is greater than 95%, the agglomeration rate is less than 10%, and the method is suitable for serum-free suspension domestication of various adherent cells and has important industrial application value.

A culture medium regeneration system is provided that allows for appropriate adjustment of the composition during the culture of cell aggregates. The culture medium regeneration system comprises: a culture medium circulation path for circulating culture medium housed in a culture section for culturing cell aggregates; a nutrient supply path for supplying regeneration culture medium containing nutrients for the cell aggregates; and a culture medium regeneration module connected to the culture medium circulation path and the nutrient supply path, and housing a semi-permeable membrane capable of allowing at least one of the nutrients and metabolites produced by the cell aggregates to pass through. The culture medium regeneration system adjusts the opportunity for the nutrients to contact the cell aggregates through the semi-permeable membrane and the opportunity for the metabolites to contact the regeneration culture medium in the nutrient supply path through the semi-permeable membrane.

This technology provides a method for accurately estimating the volume of a 3D cell aggregate from a 2D image. [Solution] An estimation method for estimating the volume of a cell aggregate, comprising: step S101 of acquiring a two-dimensional image of the cell aggregate; step S102 of extracting an object region corresponding to the cell aggregate from the two-dimensional image; step S103 of finding a reference point of the object region; step S103 of estimating a reference height which is the height of the reference point; step S105 of calculating a first distance between the reference point and the target point for each target point of the object region and a second distance between the reference point and the periphery of the object region; step S106 of calculating the height of the target point from the reference height, the first distance and the second distance; and step S107 of estimating the volume of the cell aggregate from the heights of all target points. As a result, even if the cell aggregate is deviated from a perfect sphere and the distance from the reference point to the periphery of the object region is not constant, the volume of the cell aggregate can be estimated with high accuracy.

To provide a cell aggregate capable of cryopreservation and a method for producing the cell aggregate.SOLUTION: Provided are: a cell aggregate comprising adherent cells and an extracellular matrix, the cell aggregate containing living cells even in a part located at a depth of 30 μm or more from the surface of the aggregate; and a method for producing a cell aggregate, the method comprising supporting adherent cells on a molded body formed by fibers containing an ethylene-vinylalcohol copolymer and having a non-woven fiber structure and culturing the cells under stirring.SELECTED DRAWING: None

This invention discloses a method for forming and culturing human umbilical cord mesenchymal stem cells into spheres, relating to the field of cell sphere culture technology. Specifically, it involves obtaining cell spheres using a serum-free culture medium through the following steps: selecting human umbilical cord mesenchymal stem cells and preparing a single-cell suspension; inoculating the single-cell suspension into a serum-free culture medium to obtain a cell suspension; placing the obtained cell suspension into a container and sealing it; and subjecting the container to periodic variable-speed shaking culture for 36–48 hours to form cell spheres; finally, collecting the obtained cell spheres. Compared with existing technologies, this invention effectively coordinates the cell aggregation and dispersion process by setting a specific periodic variable-speed shaking program, improving the high uniformity of the obtained cell sphere size; and throughout the entire culture process, it is serum-free, with clearly defined components, reducing the risk of batch differences and animal origin.