Screening device for analyzing multiple organoids

Abstract

A screening device for analyzing multiple organoids is disclosed. The screening device comprises: a microfluidic chip in which one or more microfluidic channels through which a drug flows are arranged; a well plate in which a plurality of chambers accommodating organoids are arranged; and a probe array in which probes inserted into the organoids so as to measure a signal in each organoid are arranged in an array form, wherein a first microfluidic channel among the one or more microfluidic channels is arranged to be adjacent to a first chamber array in which some of the chambers are arranged in an array form, and the drug flowing in the first microfluidic channel can be delivered to each organoid accommodated in the first chamber array through porous films having different porosities.

Description

Screening device for multiple organoid analysis

[0001] The disclosure below relates to a screening device for analysis of multiple organoids.

[0002] Drug screening devices measure changes in the signaling of neural organoids caused by drugs to analyze the effects of drugs and select effective ones. Neural organoids are based on human cells and have a layered structure and cellular composition similar to human organs (e.g., the brain or spinal cord). Neural organoids can be used in a wide range of applications, including drug screening, patient-specific drug selection, and organ transplantation.

[0003] In one embodiment, a screening device includes a microfluidic chip having one or more microfluidic channels through which a drug flows, a well plate having a plurality of chambers for accommodating organoids, and a probe array having probes inserted into the organoids and measuring signals within each organoid in an array form, wherein a first microfluidic channel among the one or more microfluidic channels is disposed adjacent to a first chamber array in which some of the chambers are arranged in an array form, and a drug flowing in the first microfluidic channel can be delivered to each organoid accommodated in the first chamber array through porous membranes having different porosities.

[0004] The above first chamber array includes a chamber having a porosity of 0 in the porous membrane, and a drug flowing through the first microfluidic channel may not be delivered to the organoid accommodated in the chamber having a porosity of 0.

[0005] The first chamber array includes a first chamber into which the drug is introduced by a porous membrane of a first porosity and a second chamber into which the drug is introduced by a porous membrane of a second porosity, wherein the first porosity may be different from the second porosity.

[0006] The second porosity may be greater than the first porosity and may be no greater than twice the first porosity.

[0007] The porosity of the porous membrane coupled to each chamber in the series of chamber arrays included in the first chamber array may gradually increase along the flow direction of the drug in the first microfluidic channel.

[0008] The porosity of the porous membrane coupled to each chamber in the series of chamber arrays included in the first chamber array may gradually decrease depending on the flow direction of the drug in the first microfluidic channel.

[0009] The above probes can be inserted into the organoids to measure signal changes caused by drugs delivered to each organoid.

[0010] It may further include a signal processing unit that processes signals measured by the probe array simultaneously or in parallel.

[0011] The above probe array can measure signals within organoids accommodated in a plurality of chambers arranged in an array form.

[0012] The porous membrane may be attached to the bottom surface of the well plate, and the one or more microfluidic channels may be disposed below the porous membrane.

[0013] FIG. 1 is a schematic diagram illustrating a screening device with an open cover on which a probe array is arranged according to one embodiment.

[0014] FIG. 2 is a schematic diagram showing a screening device with a closed cover having a signal processing unit disposed thereon according to one embodiment.

[0015] FIG. 3 is a drawing for explaining the difference in the concentration of a drug delivered to an organoid according to the porosity of a porous membrane according to one embodiment.

[0016] FIG. 4 is a diagram illustrating a plurality of microfluidic channels and organoids arranged in an array form adjacent to each microfluidic channel according to one embodiment.

[0017] Specific structural or functional descriptions of the embodiments are disclosed for illustrative purposes only and may be modified and implemented in various forms. Therefore, the actual implementation is not limited to the specific embodiments disclosed, and the scope of this specification includes modifications, equivalents, or alternatives within the technical concepts described in the embodiments.

[0018] Although terms such as "first" or "second" may be used to describe various components, these terms should be interpreted solely to distinguish one component from another. For example, a first component may be referred to as a second component, and similarly, a second component may also be referred to as a first component.

[0019] When it is said that a component is "connected" to another component, it should be understood that it may be directly connected or connected to that other component, but there may also be other components in between.

[0020] Singular expressions include plural expressions unless the context clearly dictates otherwise. In this document, phrases such as "A or B", "at least one of A and B", "at least one of A or B", "A, B, or C", "at least one of A, B, and C", and "at least one of A, B, or C" can each include any one of the items listed together in that phrase, or all possible combinations thereof. In this specification, it should be understood that the terms "comprises" or "has" and the like are intended to specify the presence of a described feature, number, step, operation, component, part, or combination thereof, but do not exclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0021] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art. Terms defined in commonly used dictionaries should be interpreted to have a meaning consistent with their meaning in the context of the relevant technology, and will not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0022] The term "~part" as used in this document refers to a software or hardware component such as an FPGA or ASIC, and the "~part" performs certain roles. However, the "~part" is not limited to software or hardware. The "~part" may be configured to reside on an addressable storage medium and may be configured to execute one or more processors. For example, the "~part" may include components such as software components, object-oriented software components, class components, and task components, as well as processes, functions, attributes, procedures, subroutines, segments of program code, drivers, firmware, microcode, circuits, data, databases, data structures, tables, arrays, and variables. The functionality provided within the components and "~parts" may be combined into a smaller number of components and "~parts" or further separated into additional components and "~parts." Furthermore, the components and "~parts" may be implemented to execute one or more CPUs within a device or a secure multimedia card. Additionally, '~bu' may include one or more processors.

[0023] Hereinafter, embodiments will be described in detail with reference to the attached drawings. In the description with reference to the attached drawings, identical components are assigned the same reference numerals regardless of the drawing numbers, and redundant descriptions thereof will be omitted.

[0024]

[0025] FIG. 1 is a schematic diagram illustrating a screening device with an open cover on which a probe array is arranged according to one embodiment.

[0026] The screening device described in this disclosure is a device capable of evaluating the pharmacological activity and / or toxicity of synthetic compounds or natural products that may become new drug candidates during the new drug development process. The screening device may be, for example, a drug screening device used to screen drugs, but the scope of the embodiments is not limited thereto.

[0027] The screening device may include a technology related to a multifunctional neural probe system for measuring signals of neural organoids, which is a micro electro mechanical systems (MEMS) technology. For example, the screening device may include a chamber for culturing and measuring neural organoids, a multifunctional neural probe for recording neural signals and stimulating neural cells, and a structure for controlling the position of the probe. The screening device may measure signals of the neural organoids and / or observe changes in signals (or changes in the concentration of internal substances (e.g., ions)) within the organoids due to drugs. For example, the screening device may monitor changes in pH and ion concentrations within the organoids over time and the culturing process of the organoids when the organoids are cultured. In one embodiment, the screening device may quantitatively and / or qualitatively analyze ion concentrations by inserting electrodes into immersed organoids and measuring potential differences, currents, or alternating current impedances between the electrodes. For example, a screening device can measure changes in neural signals caused by the drug by injecting a drug into multiple organoids and inserting electrodes into the immersed organoids to measure alternating current impedance between the electrodes.

[0028] The screening device used herein may include a technology for a flexible material-based three-dimensional multifunctional neural interface for functional evaluation of neural organoids. For example, the screening device may measure signals within multiple organoids or multiple spheroids. Organoids (or organ analogues) are three-dimensional multicellular or stem cell-derived micro-tissues designed to mimic human organs and may be used as analyte samples, cells, samples, or modified samples. Organoids may be replaced by analyte samples, samples, or cells. Analyte samples may refer to any sample for which ion concentrations are to be measured and may be fluids. For example, analyte samples may be saliva, urine, blood, plasma, serum, tears, pus, gastric fluid, intestinal fluid, ocular fluid, peritoneal fluid, vaginal fluid, cerebrospinal fluid, body fluid, or any combination thereof, but the subject of the assay sample is not limited thereto.

[0029] In one embodiment, a screening device can simultaneously measure signals from multiple organoids housed in a well plate using an integrated probe array. For example, the screening device can increase signal analysis throughput by simultaneously measuring brain waves from multiple organoids using an integrated neural probe array. The screening device can be used for drug screening, such as brain disease therapeutics, based on functional analysis of neural organoids, and can also be used for functional analysis of multiple organoids for functional analysis of neural organoids.

[0030] Referring to FIG. 1, the screening device may include a microfluidic chip (110) to which a first drug inlet (112), a second drug inlet (114), and a drug outlet (116) can be connected, a cover (130) that can include an array of probes (e.g., 132), and a well plate (120) in which chambers (122) that can accommodate organoids (124) are arranged.

[0031] In one embodiment, a microfluidic chip (110) may be provided with one or more microfluidic channels through which a drug flows. For example, one or more microfluidic channels through which a drug flows may be provided through a first drug inlet (112) and a second drug inlet (114) connected to the microfluidic chip (110). The drugs delivered through the first drug inlet (112) and the second drug inlet (114) according to one embodiment may be the same drug or different drugs. In one embodiment, a drug outlet (116) may be connected to the microfluidic chip (110). For example, a drug outlet (116) may be connected to discharge a drug flowing in a microfluidic channel delivered through the first drug inlet (112) and / or the second drug inlet (114). Although the drawing illustrates two drug inlets, the scope of the embodiment is not limited thereto. Depending on the embodiment, the number of drug inlets may be one or three or more.

[0032] A well plate (120) according to one embodiment may be provided with a plurality of chambers for accommodating organoids (e.g., organoids (124)). A chamber (122) according to one embodiment is a space formed to accommodate organoids (124). The plurality of chambers (122) of the well plate (120) may be arranged in an array form to accommodate a plurality of organoids (124). For example, the well plate (120) may include eight chamber arrays, each of which has twelve chambers arranged in an array form to accommodate 96 organoids (124) simultaneously.

[0033] In one embodiment, the probes may be arranged in an array form to be inserted into organoids and measure signals within each organoid. For example, the probes may be arranged in an array form corresponding to the chamber array to measure signals within organoids accommodated in the chamber array. In one embodiment, the probe array may measure signals within organoids accommodated in a plurality of chambers arranged in an array form. For example, the probe array may measure signals within organoids (124) accommodated in a chamber (122) without moving the organoids accommodated in the chamber array. In one embodiment, the probes may be inserted into organoids and measure signal changes due to drugs delivered to each organoid. For example, the probes may be inserted into organoids accommodated in the chamber array of a well plate (120), and the probes may be injected from a second drug injection port (114) and measure signal changes according to the concentration of a drug delivered to the organoid (124).

[0034]

[0035] FIG. 2 is a schematic diagram showing a screening device with a closed cover having a signal processing unit disposed thereon according to one embodiment.

[0036] Referring to FIG. 2, the screening device may include a microfluidic chip (210) connected to a first drug inlet (212), a second drug inlet (214), and a drug outlet (216), a cover (230) including a probe array, a well plate (220) having chambers capable of accommodating organoids arranged therein, and a signal processing unit (240).

[0037] The microfluidic chip (210), cover (230), and well plate (220) to which the first drug injection port (212), the second drug injection port (214), and the drug discharge port (216) of FIG. 2 can be connected correspond to the microfluidic chip (110), cover (130), and well plate (120) to which the first drug injection port (112), the second drug injection port (114), and the drug discharge port (116) of FIG. 1 can be connected, respectively, and any overlapping descriptions will be omitted below.

[0038] A screening device according to one embodiment may further include a signal processing unit (240) that processes signals measured by the probe array simultaneously or in parallel. For example, the signal processing unit (240) may be disposed on one side of the cover (230) (e.g., the upper portion outside the cover) and may perform signal processing (e.g., amplification, analog-to-digital conversion, filtering) on ​​a plurality of signals measured by the probe array within the organoids.

[0039]

[0040] FIG. 3 is a drawing for explaining the difference in the concentration of a drug delivered to an organoid according to the porosity of a porous membrane according to one embodiment.

[0041] Referring to FIG. 3, a first microfluidic channel (316) among one or more microfluidic channels may be disposed adjacent to a first chamber array in which some of the chambers among the chambers are arranged in an array form. For example, the first microfluidic channel (316) may be disposed adjacent to a first chamber array in which chambers (310, 320, 330, 340, and 350) are arranged in an array form. Each chamber of the first chamber array according to one embodiment may accommodate an organoid (e.g., 310), and probes may be inserted into each organoid. For example, a chamber (310) may accommodate an organoid (312), and a probe (314) may be inserted into an organoid (312). A screening device according to one embodiment may accommodate a plurality of organoids and process a plurality of signals simultaneously or in parallel. For example, the screening device can measure multiple brain wave signals through neural probes inserted into organoids in a 96-well plate and improve signal throughput by simultaneously processing the measured signals.

[0042] In one embodiment, a porous membrane (e.g., 321) is a membrane-like structure having multiple microscopic pores, and the porosity represents the relative ratio of the number of pores present in the porous membrane. The porous membrane may be, for example, a membrane-like structure including pores having a size of 1 μm or less, an ion exchange membrane, an epithelial tissue, or a selective ion permeable membrane having ion selectivity, but is not limited thereto. A drug flowing in a first microfluidic channel (316) according to one embodiment can be delivered to each organoid accommodated in the first chamber array through porous membranes (321, 331, 341, and 351) having different porosities. For example, a drug flowing in a first microfluidic channel (316) arranged adjacent to chambers can be delivered to each organoid accommodated in the chamber array through the porous membrane (321), the porous membrane (331), the porous membrane (341), and the porous membrane (351). In one embodiment, a porous membrane may be attached to the bottom surface of a well plate, and one or more microfluidic channels may be disposed below the porous membrane. For example, each of the porous membranes (321 to 351) may be attached to the bottom surface of a well plate (e.g., 220 of FIG. 2), and the first microfluidic channel (316) may be disposed below the porous membranes. Depending on the porosity of the porous membrane in one embodiment, the amount of drug per hour delivered to the organoids accommodated in the chamber may vary. For example, the first chamber array may include a chamber (310) to which a porous membrane having a porosity of 0 is attached, and a drug flowing in the first microfluidic channel (316) may not be delivered to the chamber (310) to which the porous membrane having a porosity of 0 is attached. According to one embodiment, the first chamber array may include a first chamber (e.g., chamber (320)) into which a drug is introduced by a porous membrane (e.g., porous membrane (321)) of a first porosity, and a second chamber (330) into which a drug is introduced by a porous membrane (e.g., porous membrane (331)) of a second porosity, wherein the first porosity may be different from the second porosity. For example, the second porosity may be greater than the first porosity and no greater than twice the first porosity.In one embodiment, a microfluidic chip can deliver various types of drugs through each microfluidic channel, and a porous membrane is attached to the bottom of a well plate to control the concentration of the drug delivered according to the porosity of the porous membrane. For example, the microfluidic chip can deliver different concentrations of drugs to each organoid accommodated in a chamber array through a porous membrane (311) with a porosity of (0%), a porous membrane (321) with a porosity of (1.25%), a porous membrane (331) with a porosity of (2.5%), a porous membrane (341) with a porosity of (3.75%), and a porous membrane (351) with a porosity of (5%).

[0043]

[0044] FIG. 4 is a diagram illustrating a plurality of microfluidic channels and organoids arranged in an array form adjacent to each microfluidic channel according to one embodiment.

[0045] A microfluidic channel is a channel that can flow a trace amount of analyte formed in a microfluidic chip. One end of a microfluidic channel (430) according to one embodiment may be connected to a drug inlet and the other end may be connected to a drug outlet. For example, one end of a first microfluidic channel (430) may be connected to a second drug inlet (114) and the other end may be connected to a drug outlet (116), and one end of a second microfluidic channel (420) may be connected to the first drug inlet (112) and the other end may be connected to a drug outlet (116).

[0046] Referring to FIG. 4, drawing (410) illustrates microfluidic channels arranged adjacent to each chamber array. According to one embodiment, a first microfluidic channel (430) may be arranged adjacent to the first chamber array (431 to 435), and a second microfluidic channel (420) may be arranged in the second chamber array (421 to 425). Drawing (440) according to one embodiment illustrates an enlarged view of a chamber (435) arranged adjacent to the first microfluidic channel (430). A drug flowing in the first microfluidic channel (430) according to one embodiment may be delivered to each organoid accommodated in the first chamber array (431 to 435) through the porous membrane. According to one embodiment, the first microfluidic channel (430) may correspond to the first microfluidic channel (444) of drawing (440). According to one embodiment, a drug flowing in the first microfluidic channel (444) can be delivered to an organoid (443) accommodated in a chamber (442) through a porous membrane (445), and the probe (441) can measure a signal by the delivered drug. The flow direction (450) of the drug flowing in the first microfluidic channel (430) according to one embodiment may be to the right when viewed from the front in FIG. 4. According to one embodiment, a drug flowing in the first microfluidic channel (430) can be delivered to each organoid accommodated in the first chamber array through porous membranes having different porosities.

[0047] In a series of chamber arrays included in a first chamber array according to one embodiment, the porosity of the porous membrane attached to each chamber may gradually increase along the flow direction (450) of the drug in the first microfluidic channel. When the porosity of the porous membrane attached to each chamber gradually increases along the flow direction (450) of the drug, the concentration of the drug delivered to the organoids accommodated in the chambers arranged along the flow direction (450) of the drug may increase. For example, the concentration of the drug delivered to the chamber (435) (e.g., 5%) may be greater than the concentration of the drug delivered to the chamber (432) (e.g., 1.25%).

[0048] In one embodiment, the porosity of the porous membrane coupled to each chamber in a series of chamber arrays included in the first chamber array may gradually decrease depending on the flow direction of the drug in the first microfluidic channel. When the porosity of the porous membrane coupled to each chamber gradually decreases depending on the flow direction of the drug (450), the concentration of the drug delivered to the organoids accommodated in the chambers arranged depending on the flow direction of the drug (450) may decrease. For example, the concentration of the drug delivered to the chamber (435) (e.g., 2.5%) may be lower than the concentration of the drug delivered to the chamber (433) (e.g., 5%).

[0049] The screening device can detect the effectiveness of a drug by delivering the drug to a chamber containing organoids through a microfluidic channel for drug delivery. The screening device can identify the optimal drug concentration without multiple experiments by observing signal changes according to drug concentration. The screening device has microfluidic channels attached to the bottom of each well plate containing organoids for drug delivery of different concentrations, enabling the identification of highly efficacious drugs and optimal drug concentrations.

[0050] A screening device according to one embodiment measures organoid signals simultaneously or in parallel and has a drug screening function based on drug concentration. A screening device capable of simultaneously or in parallel measuring organoid signals can be widely used in functional analysis of brain disease organoids and screening for brain disease therapeutics.

[0051]

[0052] The hardware devices described above may be configured to operate as one or more software modules to perform the operations of the embodiments, and vice versa.

[0053] Although the embodiments described above have been described with limited drawings, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the described embodiments. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0054] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. In the screening device, A microfluidic chip having one or more microfluidic channels through which a drug flows; A well plate having multiple chambers arranged to accommodate organoids; and A probe array in which probes are inserted into the above organoids and arranged in an array form to measure signals within each organoid. Including, A first microfluidic channel among the one or more microfluidic channels is disposed adjacent to a first chamber array in which some of the chambers among the chambers are arranged in an array form, The drug flowing in the first microfluidic channel is delivered to each organoid accommodated in the first chamber array through porous membranes having different porosities. Screening device.

2. In paragraph 1, The above first chamber array includes a chamber having a porosity of 0 in the porous membrane, In the chamber with the above porosity of 0, the drug flowing in the first microfluidic channel is not delivered to the organoid contained in the chamber with the above porosity of 0. Screening device.

3. In paragraph 1, The above first chamber array, It includes a first chamber into which the drug is injected through a porous membrane of a first porosity and a second chamber into which the drug is injected through a porous membrane of a second porosity, The above first porosity is different from the above second porosity, Screening device.

4. In paragraph 3, The above second porosity is, A porosity greater than the first porosity and less than twice the first porosity, Screening device.

5. In paragraph 1, The porosity of the porous membrane coupled to each chamber in the series of chamber arrays included in the first chamber array gradually increases along the flow direction of the drug in the first microfluidic channel. Screening device.

6. In paragraph 1, The porosity of the porous membrane coupled to each chamber in the series of chamber arrays included in the first chamber array gradually decreases along the flow direction of the drug in the first microfluidic channel. Screening device.

7. In paragraph 1, The above probes are, Inserted into the above organoids and measuring the signal change due to the drug delivered to each organoid Screening device.

8. In paragraph 1, A signal processing unit that processes signals measured by the above probe array simultaneously or in parallel. Including more Screening device.

9. In paragraph 1, The above probe array, Measuring signals within organoids housed in multiple chambers arranged in an array form. Screening device.

10. In paragraph 1, The above porous membrane is attached to the bottom surface of the well plate, wherein said one or more microfluidic channels are disposed below said porous membrane, Screening device.

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