KR102691068B1 - Apparatus for brain neural modeling - Google Patents

Abstract

A brain nerve modeling device that simulates the microenvironment of the brain according to an embodiment is disclosed. The brain nerve modeling device is a microfluidic chip including a plurality of cell culture channels and electrodes for applying an electrical stimulation signal to at least one of the cell culture channels or measuring an activity signal of nerve cells cultured in the cell culture channels. , a control unit that determines the electrical stimulation signal to be provided through the electrodes and processes the measured activity signal, and a micro pump that provides culture medium to the cell culture channels, and the cell culture channels are a first cerebrovascular gate simulation channel, a nerve It may include a glial cell channel, a first neuron channel, an axonal guidance channel, a second neuron channel, and a second cerebrovascular portal mimic channel.

Description

Brain nerve modeling device {APPARATUS FOR BRAIN NEURAL MODELING}

The following embodiments relate to a brain nerve modeling device and control technology therefor.

Cell culture technology using microfluidic technology is attracting attention in various fields because it can more accurately reflect actual in vivo mechanisms by providing a microenvironment suitable for cells by simulating the human body and implementing their functions and characteristics. Recently, lab on a chip technology using microfluidic technology has been used in the field of neuroscience. By using human-derived cells, lab-on-a-chip technology can complement the limitations of studies on the development and progression of neurological disorders conducted through existing animal experiments. In lab-on-a-chip technology, the movement and absorption of oxygen and nutrients and signal transmission processes between tissues or cells within the actual human body are important, so brain tissue layers composed of various cells such as nerve cells, glial cells, endothelial cells, and epithelial cells are simulated. Multiple cells can be co-cultured. However, current technology has limitations in imitating all of the complex systems of the human body, so a comprehensive platform is needed to understand the mechanisms of specific cells.

A brain nerve modeling device that simulates the microenvironment of the brain according to one embodiment includes a plurality of cell culture channels and applying an electrical stimulation signal to at least one of the cell culture channels or nerve cells cultured in the cell culture channels. A microfluidic chip including electrodes for measuring activation signals of; a control unit that determines an electrical stimulation signal to be provided through the electrodes and processes the measured activation signal; And a micro pump that provides a culture medium to the cell culture channels, wherein the cell culture channels include a first cerebrovascular portal mimic channel, a glial cell channel, a first nerve cell channel, an axonal guidance channel, and a second nerve cell channel. It may include a channel and a second cerebrovascular portal simulating channel.

Neuronal axons of nerve cells cultured in the first nerve cell channel may be cultured in a serial form through the axon guidance channel and form a synapse with the second nerve cell channel.

The microfluidic chip includes posts forming an interface between the plurality of cell culture channels, and a gap between the posts may be smaller than the height of the plurality of cell culture channels.

The microfluidic chip forms an interface based on the posts arranged at regular intervals, and at the interface, movement of fluid is prevented and exchange of biochemical substances between the cell culture channels can occur.

The microfluidic chip further includes a cover slip on which the electrodes are patterned, the electrodes are connected to the control unit through a connector, and can measure activity signals of nerve cells cultured in the cell culture channels.

The cover slip is placed below the microfluidic chip, and cells can be cultured by attaching to the cover slip.

The control unit includes a power unit that provides power to the control unit; an amplifying unit that amplifies the measured activation signal; And it may include a controller unit that controls at least one of the power supply unit and the amplifier unit.

The micropump may circulate the culture medium in a single direction through the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel.

The micropump may circulate the culture medium in the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel through an open loop circulation method or a closed loop circulation method.

Each of the cell culture channels includes an injection port for injecting nerve cells; and an outlet, wherein the inlet and the outlet are formed by perforating the microfluidic chip, and the culture medium can be circulated through the inlet and outlet.

The microfluidic chip can independently co-culture nerve cells in each of the cell culture channels.

According to one embodiment, the limitations of conventional animal experiments and brain-simulating chips can be overcome.

According to one embodiment, cells are co-cultured differently in each culture channel using microfluidic technology, and through this, a microenvironment that mimics the human brain microenvironment can be constructed.

According to one embodiment, low flow rate cerebral blood flow circulating in a single direction can be simulated through dynamic culture using a micropump.

According to one embodiment, various co-culture conditions, as well as drug and electrical tests, are possible, so the interaction between cells for each test can be effectively evaluated.

According to one embodiment, neurodegenerative diseases can be more intuitively simulated through the structure of a plurality of differentiated nerve cell culture channels and the nerve axon channel structure connecting them.

According to one embodiment, electrical stimulation can be applied and sensed through a microelectrode, making it possible to evaluate electrical stimulation treatment, measure activity signals of nerve cells, or evaluate synapses between cells.

According to one embodiment, since it has a transparent chip structure, it is possible to observe phenotypic changes in a simulated microenvironment in real time.

Figures 1 and 2 are diagrams showing the structure of a microfluidic chip according to one embodiment.
FIG. 3 is a diagram illustrating cell culture channels of a microfluidic chip simulating the human body according to an embodiment.
FIG. 4 is a diagram illustrating a synapse formed between nerve cell channels according to an embodiment.
FIGS. 5 and 6 are diagrams for explaining a microfluidic chip and a substrate according to an embodiment.
FIG. 7 is a diagram illustrating the configuration of a brain nerve modeling apparatus according to an embodiment.

Specific structural or functional descriptions of the embodiments are disclosed for illustrative purposes only and may be changed and implemented in various forms. Accordingly, the actual implementation form is not limited to the specific disclosed embodiments, and the scope of the present specification includes changes, equivalents, or substitutes included in the technical idea described in the embodiments.

Terms such as first or second may be used to describe various components, but these terms should be interpreted only for the purpose of distinguishing one component from another component. For example, a first component may be named a second component, and similarly, the second component may also be named a first component.

When a component is referred to as being “connected” to another component, it should be understood that it may be directly connected or connected to the other component, but that other components may exist in between.

Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as “comprise” or “have” are intended to designate the presence of the described features, numbers, steps, operations, components, parts, or combinations thereof, and are intended to indicate the presence of one or more other features or numbers, It should be understood that this does not exclude in advance the possibility of the presence or addition of steps, operations, components, parts, or combinations thereof.

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 as defined in commonly used dictionaries should be interpreted as having meanings consistent with the meanings they have in the context of the related technology, and unless clearly defined in this specification, should not be interpreted in an idealized or overly formal sense. No.

Hereinafter, embodiments will be described in detail with reference to the attached drawings. In the description with reference to the accompanying drawings, identical components will be assigned the same reference numerals regardless of the reference numerals, and overlapping descriptions thereof will be omitted.

The brain nerve modeling device described in this specification may be intended to solve the limitations of conventional animal experiments and chips that simulate the brain. A brain nerve modeling device can simulate degenerative neurological diseases and can be a platform to more accurately study and evaluate in vivo mechanisms using co-culture technology and microelectrodes.

Recently, as the importance of the role of glial cells in neurodegenerative diseases has emerged, methods to evaluate the responsiveness of nerve cells and the efficacy and toxicity of drugs through interactions between co-cultured cells are being studied.

The brain nerve modeling device described in this specification can effectively evaluate interactions between nerve cells for various experiments in an environment similar to the brain microenvironment by simulating the human brain microenvironment.

The cranial nerve modeling device can generate a cranial nerve model modeling cranial nerves based on microfluidic technology. The brain nerve model can use microscopic electrodes as real-time biosensors. For example, the electrode can measure the permeability of the blood-brain barrier (BBB) using TEER (Trans-epithelial electrical resistance) measurement, measure the activity signal of nerve cells, or apply an electrical stimulation signal. can do. The TEER measurement method can be a measurement method that measures how much tight junctions have been formed. In other words, the TEER measurement method may be a measurement method that measures the degree to which tight junctions are formed.

The brain nerve modeling device described herein may include a microfluidic chip, a control unit, and a micropump. The control unit may determine an electrical stimulation signal to be provided through the electrodes of the microfluidic chip and process the measured activation signal. Additionally, the micropump can provide culture medium to cell culture channels. Description of the control unit and micro pump can be explained in more detail in FIG. 7.

The microfluidic chip may include a plurality of cell culture channels and electrodes for applying an electrical stimulation signal to at least one of the cell culture channels or measuring an activity signal of cells cultured in the cell culture channels. The microfluidic chip can co-culture nerve cells independently in each of the cell culture channels.

Figures 1 and 2 are diagrams showing the structure of a microfluidic chip according to one embodiment. Referring to FIG. 1, the microfluidic chip 120 may include an electrode 110 and a plurality of cell culture channels. The cell culture channels included in the microfluidic chip 120 are a first cerebrovascular portal simulating channel 130, a glial cell channel 150, a first nerve cell channel 160, an axon guidance channel 170, and a second It may include a nerve cell channel 180 and a second cerebrovascular portal simulating channel 190.

The microfluidic chip 120 may also include posts 140 that form an interface between a plurality of cell culture channels. The spacing between the posts 140 may be smaller than the height of the plurality of cell culture channels to prevent fluid movement between the cell culture channels. The microfluidic chip 120 may form an interface based on the posts 140 arranged at regular intervals. For example, each cell culture channel may have a width of 1000 μm and a height of 250 μm, and an interface may be formed between each cell culture channel based on posts 140 spaced at 100 μm intervals. At the interface, movement of fluid is prevented and exchange of biochemical substances between cell culture channels can occur.

Neuroglial cell channel 150 may include glial cells. Neuroglial cells are one-tenth the size of nerve cells, but exist in about 10 times the number, and can perform nerve cell recovery, insulation, immunity, nutrient supply, and regulation of extracellular fluid.

The first nerve cell channel 160 can culture nerve cells. Neuronal axons of nerve cells cultured in the first nerve cell channel 160 are cultured in a serial form through the axon guidance channel 170, and may form a synapse with the second nerve cell channel 180. Through this, the microfluidic chip 120 can grow nerve axons in an aligned form.

The microfluidic chip 120 is capable of physically or chemically simulating a neurodegenerative disease, and glial cells in at least one of the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel or glial cell channel. By controlling cells, it is possible to enable research under various conditions.

Referring to FIG. 2, the microfluidic chip may further include a cover slip on which electrodes 210, 215, 220, and 225 are patterned. Among the electrodes, electrodes 210 and 215 may be connected to the control unit through a connector. Additionally, the electrodes 220 and 225 may be disposed in the first nerve cell channel and the second nerve cell channel to apply electrical stimulation signals or measure activity signals of nerve cells cultured in the cell culture channels. Additionally, the electrodes 220 and 225 may apply or measure various signals. The electrodes 210, 215, 220, and 225 may be made of materials with excellent biocompatibility, for example, titanium nitride (TiN) or indium tin oxide (ITO). The size of the electrodes 210, 215, 220, and 225 may be, for example, about 30 μm in diameter and 200 μm in spacing to enable stimulation and activity measurement at the nerve cell level.

In one embodiment, each of the cell culture channels may include an inlet 230 and an outlet 240 for injecting nerve cells. The inlet 230 and outlet 240 may be formed by perforating a microfluidic chip. The culture medium may be circulated through the inlet 230 and outlet 240. In this embodiment, nerve cells injected through the injection port 230 may circulate in the direction of the outlet port 240.

In another embodiment, each of the cell culture channels may include an inlet 240 and an outlet 230. The culture medium may be circulated through the inlet 240 and outlet 230. In this embodiment, nerve cells injected through the inlet 240 may circulate in the direction of the outlet 230.

FIG. 3 is a diagram illustrating cell culture channels of a microfluidic chip simulating the human body according to an embodiment.

Referring to Figure 3, a microfluidic chip can simulate the microenvironment of the brain. The microfluidic chip may include a plurality of cell culture channels, and among the cell culture channels, the cerebrovascular portal-mimetic channel may include vascular endothelial cells that simulate blood vessels of the human body. Additionally, the glial cell channel may include glial cells 320 that mimic human glial cells, and the nerve cell channel may include nerve cells 330 that mimic human neuroglial cells.

FIG. 4 is a diagram illustrating a synapse formed between nerve cell channels according to an embodiment.

Referring to FIG. 4, step 410 represents the form before the nerve axon 440 cultured in the first nerve cell channel 450 forms a synapse with the second nerve cell channel 470, and step 420 represents the form before forming a synapse with the second nerve cell channel 470. The shape of the nerve axon 442 cultured in the first nerve cell channel 450 may be shown after forming a synapse with the second nerve cell channel 470.

Referring to step 410, the first nerve cell channel 450 and the second nerve cell channel 470 may include nerve cells, and the first nerve cell channel 450 and the second nerve cell channel 470 ) The electrode 430 may be disposed. In step 410, even if an electrical stimulation signal is applied to the first nerve cell channel 450 by the electrode 430, the electrical stimulation signal may not be measured in the second nerve cell channel 470. If the nerve axon 440 is cultured along the axon guidance channel 460 in step 410, the nerve axon 442 may be connected to a nerve cell included in the second nerve cell channel 470 in step 420. At this time, the electrical stimulation signal applied by the electrode 430 of the first nerve cell channel 450 may also be transmitted to the electrode 480 disposed in the second nerve cell channel 470 along the nerve axon. Electrical stimulation signals can also be measured in nerve cell channels included in the nerve cell channel 470.

FIGS. 5 and 6 are diagrams for explaining a microfluidic chip and a substrate according to an embodiment.

Referring to FIG. 5, the microfluidic chip 520 may include a cover slip on which electrodes are patterned. A cover slip can be placed under the microfluidic chip 520, and the microfluidic chip 520 can be combined with the substrate 510 to complete the platform. For example, the microfluidic chip 520 may be composed of a silicon wafer master manufactured through a photolithography or soft-lithography process. Additionally, the microfluidic chip 520 may be made of materials such as polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA), polyacrylate, and glass, but is not limited thereto. Additionally, the cover slip may be made of a transparent material such as glass.

Referring to Figure 6, cells can be cultured attached to the cover slip 630. To facilitate applying electrical stimulation signals to nerve cells and measuring activity signals of nerve cells, cells can be cultured attached to the bottom of the cover slip 630. The cover slip 630 may have electrodes 640 patterned, and the microfluidic chip may include an inlet 610 and an outlet 620 formed by perforating the microfluidic chip. The culture medium may be circulated through the inlet 610 and outlet 620 and provided to the nerve cells.

FIG. 7 is a diagram illustrating the configuration of a brain nerve modeling apparatus according to an embodiment.

Referring to FIG. 7 , the brain nerve modeling device may include a control unit 710, a connector 750, a microfluidic chip 760, and a micro pump 770.

In one embodiment, the microfluidic chip 760 includes a plurality of cell culture channels and an electrode for applying an electrical stimulation signal to at least one of the cell culture channels or measuring an activity signal of nerve cells cultured in the cell culture channels. may include. The microfluidic chip 760 may further include a cover slip on which electrodes are patterned. The electrodes are connected to the control unit 710 through the connector 750, and can measure the activity of nerve cells cultured in the cell culture channels.

The control unit 710 may determine electrical stimulation signals to be provided through the electrodes and control the electrodes to measure or sense the activity signals of nerve cells. The control unit 710 may process the measured activation signal. The control unit 710 may include a power unit 730, an amplifier 720, and a controller unit 740. The power unit 730 may provide power to the control unit 710. The amplification unit 720 may amplify the measured activation signal. The controller unit 740 can control at least one of the power supply unit 730 and the amplifier unit 720. The controller unit 740 may also be referred to as a microcontroller unit or MCU (micro controller unit).

The micro pump 770 can provide culture medium to the cell culture channels. The micropump 770 may circulate the culture medium in a single direction through the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel. Additionally, the micropump 770 may circulate the culture medium to the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel through an open-loop circulation method or a closed-loop circulation method. The mode of circulation of the culture medium may be determined based on the experimental design. For example, the micropump 770 can simulate cerebral blood flow by circulating culture medium at a low flow rate of 0.1 dyn/cm.

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

As described above, although the embodiments have been described with limited drawings, those skilled in the art can apply various technical modifications and variations based on this. For example, the described techniques are performed in a different order than the described method, and/or components of the described system, structure, device, circuit, etc. are combined or combined in a different form than the described method, or other components are used. Alternatively, appropriate results may be achieved even if substituted or substituted by an equivalent.

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

110, 210, 215, 220, 225, 430, 480, 640: Electrodes
120, 520, 760: Microfluidic chip 140: Post
510: substrate 610: injection port
620: outlet 710: control unit
630: cover slip 720: amplification unit
730: Power unit 740: Controller unit
750: Connector 770: Micro Pump

Claims (11)

In a brain nerve modeling device that simulates the microenvironment of the brain,
A microfluidic chip including a plurality of cell culture channels and electrodes for applying an electrical stimulation signal to at least one of the cell culture channels or measuring an activity signal of nerve cells cultured in the cell culture channels;
a control unit that determines an electrical stimulation signal to be provided through the electrodes and processes the measured activation signal; and
It includes a micro pump that supplies culture medium to the cell culture channels,
The cell culture channels are,
It includes a first cerebrovascular portal-mimetic channel, a glial cell channel, a first neuronal channel, an axonal guidance channel, a second neuronal cell channel, and a second cerebrovascular portal-mimetic channel;
Neuronal axons of nerve cells cultured in the first nerve cell channel,
Cultured in serial form through the axon guidance channel, forming a synapse with the second nerve cell channel,
Brain nerve modeling device. delete In a brain nerve modeling device that simulates the microenvironment of the brain,
A microfluidic chip including a plurality of cell culture channels and electrodes for applying an electrical stimulation signal to at least one of the cell culture channels or measuring an activity signal of nerve cells cultured in the cell culture channels;
a control unit that determines an electrical stimulation signal to be provided through the electrodes and processes the measured activation signal; and
It includes a micro pump that supplies culture medium to the cell culture channels,
The cell culture channels are,
It includes a first cerebrovascular portal-mimetic channel, a glial cell channel, a first neuronal channel, an axonal guidance channel, a second neuronal cell channel, and a second cerebrovascular portal-mimetic channel;
The microfluidic chip,
Includes posts forming an interface between the plurality of cell culture channels,
The spacing between the posts is,
Smaller than the height of the plurality of cell culture channels,
Brain nerve modeling device. According to paragraph 3,
The microfluidic chip forms an interface based on the posts arranged at regular intervals,
At the interface, movement of fluid is prevented and exchange of biochemical substances between the cell culture channels occurs,
Brain nerve modeling device. In a brain nerve modeling device that simulates the microenvironment of the brain,
A microfluidic chip including a plurality of cell culture channels and electrodes for applying an electrical stimulation signal to at least one of the cell culture channels or measuring an activity signal of nerve cells cultured in the cell culture channels;
a control unit that determines an electrical stimulation signal to be provided through the electrodes and processes the measured activation signal; and
It includes a micro pump that supplies culture medium to the cell culture channels,
The cell culture channels are,
It includes a first cerebrovascular portal-mimetic channel, a glial cell channel, a first neuronal channel, an axonal guidance channel, a second neuronal cell channel, and a second cerebrovascular portal-mimetic channel;
The microfluidic chip,
Further comprising a cover slip on which the electrodes are patterned,
The electrodes are,
Connected to the control unit through a connector and measuring activity signals of nerve cells cultured in the cell culture channels,
Brain nerve modeling device. According to clause 5,
The cover slip is placed below the microfluidic chip,
Cells are cultured by attaching to the cover slip,
Brain nerve modeling device. According to paragraph 1,
The control unit,
a power supply unit that provides power to the control unit;
an amplifying unit that amplifies the measured activation signal; and
A controller unit that controls at least one of the power supply unit and the amplifier unit.
Including,
Brain nerve modeling device. According to paragraph 1,
The micro pump,
Circulating the culture medium in a single direction in the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel,
Brain nerve modeling device. According to clause 8,
The micro pump,
Circulating the culture medium in the first cerebrovascular portal simulating channel and the second cerebrovascular portal simulating channel through an open loop circulation method or a closed loop circulation method,
Brain nerve modeling device. According to paragraph 1,
Each of the cell culture channels,
An injection port for injecting nerve cells; and
outlet
Including,
The inlet and the outlet are,
The microfluidic chip is formed by perforating,
The culture medium is circulated through the inlet and outlet,
Brain nerve modeling device. According to paragraph 1,
The microfluidic chip,
Co-culturing nerve cells independently in each of the cell culture channels,
Brain nerve modeling device.

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