Long-term culture device and system for brain organs

Abstract

The utility model provides a brain organ long-term culture device and a brain organ long-term culture system. The brain organ long-term culture device comprises an incubator, a brain organ chip, a microelectrode array and a microscopic imaging module, the culture box provides a culture environment for brain organs in the brain organ chip; the brain organ chip comprises light-permeable glass located on the upper layer and a base layer located on the lower layer. The center of the base layer sinks to form a container which is communicated with the outside of the chip; the space of the container is divided into two parts by a semi-permeable membrane, and a brain organ matrigel mixture is placed above the semi-permeable membrane; the microelectrode array tightly covers the brain organ to collect electroencephalogram signals; the microscopic imaging module is used for collecting images of brain organs on the brain organ chip. According to the device, a micro-fluidic chip technology is adopted, automatic culture of brain organs is achieved, waste of culture media is reduced, manpower and material resources are saved, standardization and batch culture of the brain organs are better achieved, and brain scientific research is efficiently assisted.

Description

Technical Field

[0001] This invention relates to the field of bioengineering culture equipment, and more particularly to a long-term brain organoid culture device and system. Background Technology

[0002] Brain organoids are three-dimensional, brain-like tissues produced by the differentiation of induced pluripotent stem cells (iPSCs) in a culture medium that simulates the brain's developmental environment. Brain organoids can be used to study development, diseases, or drug effects, providing unprecedented opportunities for researching and understanding brain function and the pathogenesis of neurological diseases. Traditional brain organoid manipulation involves numerous tedious steps and manual operations, consumes large amounts of culture medium, and faces challenges in standardization and mass production.

[0003] Today, microfluidic technology can be used to integrate organ culture into microfluidic chips, enabling automated organ culture while precisely controlling the culture microenvironment, reducing culture medium waste, saving manpower and resources, and facilitating standardized and mass culture of brain organoids, thus efficiently supporting brain science research.

[0004] Therefore, providing a long-term culture device for brain organoids based on microfluidics technology is a noteworthy need. Utility Model Content

[0005] The present invention aims to provide a long-term culture device for brain organoids based on microfluidic technology, the technical solution of which is as follows:

[0006] A long-term brain organoid culture device, characterized in that it comprises: an incubator, a brain organoid chip, a microelectrode array, and a microscopic imaging module; the incubator is used to provide a culture environment for brain organoids in the brain organoid chip;

[0007] The brain organoid chip comprises: a light-transmitting glass layer on top and a base layer on the bottom; a container is formed in the center of the base layer, the upper edge of the container is connected to the outside of the chip through an inlet pipe, and the bottom of the container is connected to the outside of the chip through an outlet pipe; the space of the container is divided into two parts by a semi-permeable membrane, the bottom of the container is used to store culture medium, and the area above the semi-permeable membrane is used to place a brain organoid matrix gel mixture, the semi-permeable membrane is used for the exchange of nutrients between the brain organoid matrix gel mixture and the culture medium;

[0008] The microelectrode array is closely covered above the brain organoid matrix gel mixture and is led out through the EEG signal transmission line for collecting EEG signals;

[0009] The microscopic imaging module is used to acquire images of brain organoids on the brain organoid chip.

[0010] Furthermore, the container is a cylindrical container, and the semi-permeable membrane covers the container and sinks to form a hemispherical space.

[0011] Furthermore, the brain organoid chip has a detachable structure, with the four corners of its upper and lower layers fixed with screws.

[0012] Furthermore, the microelectrode array adopts a flexible, rhomboid mesh structure.

[0013] Furthermore, the microscopic imaging module employs a macro camera with a focal length of 50mm to 100mm.

[0014] Furthermore, the incubator includes a chamber, a control module, and a sterilization module, a temperature module, a humidity module, and a carbon dioxide concentration monitoring module electrically connected to the control module.

[0015] Furthermore, the sterilization module is an ultraviolet lamp.

[0016] Furthermore, the incubator is also equipped with an oscillation device for oscillating the brain organoid chip.

[0017] Furthermore, the oscillation device is an oscillation table or a vibrating arm.

[0018] Furthermore, the brain organoid chip is disposed on the oscillator or connected to the oscillator.

[0019] This invention also provides an automatically controllable long-term culture system suitable for brain organoids, the technical solution of which is as follows:

[0020] A long-term brain organoid culture system includes a culture medium, a waste liquid storage device, an electroencephalogram (EEG) signal processor, and the brain organoid long-term culture device as described above. The culture medium is connected to the inlet pipe of the brain organoid long-term culture device for providing culture medium, and the waste liquid storage device is connected to the outlet pipe of the brain organoid long-term culture device for collecting waste liquid. Both the inlet pipe and the outlet pipe are equipped with microfluidic control valves. The microelectrode array is connected to the EEG signal processor via an EEG signal transmission line.

[0021] Technical effects:

[0022] This device uses microfluidic chip technology to achieve automated culture of brain organoids while precisely controlling the culture microenvironment, reducing culture medium waste, saving manpower and resources, and making it more conducive to the standardized and batch culture of brain organoids, thus efficiently supporting brain science research. Attached Figure Description

[0023] Figure 1 is a brain organ long-term culture device and system block diagram of the present application.

[0024] 1: constant temperature refrigerator; 2: culture medium; 3: liquid inlet pipe; 4: microfluidic control valve; 5: incubator; 6: sterilization module; 7: temperature module; 8: microscopic imaging module; 9: humidity module; 10: brain organ chip; 11: culture solution; 12: semi-permeable membrane; 13: brain organ matrigel mixture; 14: oscillation device; 15: liquid outlet pipe; 16: waste liquid storage device; 17: microelectrode array; 18: electroencephalogram transmission line; 19: electroencephalogram processor; 20: data transmission line; 21: data analysis and processing shared platform. DETAILED DESCRIPTION

[0025] To further illustrate the embodiments, the present application provides drawings. These drawings are part of the disclosure of the present application, which mainly serve to illustrate the embodiments, and can be explained in conjunction with the related description of the specification to understand the operating principle of the embodiments. With reference to these contents, those skilled in the art should understand other possible implementations and advantages of the present application. The components in the drawings are not drawn to scale, and similar component symbols are generally used to represent similar components.

[0026] The present application will be further described in conjunction with the drawings and specific embodiments.

[0027] As Figure 1 shown, the utility model discloses a brain organ long-term culture system, which is composed of a culture medium 2, a brain organ long-term culture device, an electroencephalogram processing system and a waste liquid storage device 16.

[0028] The culture medium 2 is stored in a constant temperature refrigerator 1 at 4 DEG C, so that long-term temperature storage of the culture medium 2 can be realized. The culture medium 2 flows into the brain organ long-term culture device through a liquid inlet pipe 3, and manual replacement is required for switching different culture media 2. The waste liquid generated during the culture of the brain organ long-term culture device is discharged into a waste liquid treatment device through a liquid outlet pipe 15 for centralized treatment. The liquid inlet pipe 3 and the liquid outlet pipe 15 are provided with a microfluidic control valve 4, so that the speed of the culture medium flowing in can be monitored and adjusted. The above-mentioned modules and devices form a microfluidic perfusion and discharge system, which can control the long-term automatic culture microenvironment of the brain organ.

[0029] The long-term culture device of the brain organoid includes a culture box 5, a microelectrode array 17, and a brain organoid microfluidic chip (hereinafter referred to as a brain organoid chip 10), etc. The culture box 5 is a container for culturing the brain organoid, and can accommodate multiple brain organoid chips 10 at the same time. The culture box 5 includes a box body and a control module, and is connected with a sterilization module 6, a temperature module 7, a humidity module 9, a carbon dioxide concentration monitoring module (i.e. a carbon dioxide sensor, not shown), a microscopic imaging module 8, and an oscillation device 14, etc. The culture box 5 has the functions of sterilization, constant temperature control, humidity control, carbon dioxide concentration control, and real-time image acquisition, etc. The brain organoid can be cultured in a constant temperature, constant humidity, sterile environment, and the growth state of the brain organoid can be monitored at the same time.

[0030] Optionally, the sterilization module 6 adopts an ultraviolet lamp. The culture box 5 ensures the sterility of the culture environment by installing an ultraviolet lamp.

[0031] Optionally, the temperature module 7 is a constant temperature control system, which is composed of a temperature sensor, a temperature controller, a heating device (such as a PTC heating sheet), and a refrigeration device (such as a semiconductor refrigeration sheet), etc. Through temperature detection and control, the culture box 5 can maintain an appropriate environment of 37℃.

[0032] Optionally, the humidity module 9 is composed of a humidity sensor and a dehumidifier, etc. Through humidity monitoring and control, the culture box 5 can maintain an appropriate humidity.

[0033] Optionally, the microscopic imaging module 8 adopts a macro camera, and the focal length thereof is preferably 50mm to 100mm. The brain organoid on the brain organoid chip 10 can be imaged in real time to monitor the growth state of the brain organoid.

[0034] The oscillation device 14 is connected with the brain organoid chip 10, and can oscillate the brain organoid chip 10 through the oscillation device 14, so that the brain organoid can grow better. Optionally, the oscillation device 14 can be an oscillation table or an oscillation arm, which is arranged at the bottom or one side of the brain organoid chip 10.

[0035] In the embodiment, the brain organoid chip 10 is a two-layer sandwich structure, the upper layer is a light-transmissive glass, and the lower layer is a polyethylene base layer, and the central part of the base layer is sunken to form a container. The upper edge of the container is connected to the outside of the chip through a liquid inlet pipe 3, and the culture solution 11 is injected. The bottom of the container is connected to the outside through a liquid outlet pipe 15, and the waste liquid generated during the culture process is discharged. The liquid inlet pipe 3 is arranged between the upper and lower layers of the brain organoid chip 10. Optionally, the container is a cylindrical container. The cylindrical container is covered with a semi-permeable membrane 12, and the space of the cylindrical container is divided into two parts by the semi-permeable membrane 12: the culture solution is stored at the bottom of the container, and the semi-permeable membrane 12 is a semispherical brain organoid matrigel mixture 13 above the semi-permeable membrane 12. The brain organoid matrigel mixture 13 and the culture solution exchange nutrients through the semi-permeable membrane 12. The brain organoid chip 10 is a detachable structure, and the four corners of the upper and lower layers of the brain organoid chip 10 are fixed by screws.

[0036] In the embodiment, the brain organoid matrigel mixture 13 is closely covered with a flexible high-density microelectrode array 17. Optionally, the microelectrode array 17 adopts a rhombic net-shaped microelectrode array, which can provide N*M electrodes and is made of platinum. The microelectrode array 17 is used to collect the electrical signals of the brain organoid, and is connected to a brain electrical signal processing system through a bundle of brain electrical signal transmission lines 18. In the embodiment, the brain electrical signal processing system includes a brain electrical signal processor 19 and a data analysis processing shared platform 21. The microelectrode array 17 is connected to the brain electrical signal processor through the brain electrical signal transmission lines 18, and the brain electrical signal processor is connected to the data analysis processing shared platform 21 through a data transmission line 20 such as a network cable. Optionally, the brain electrical signal processor 19 can perform preliminary processing such as filtering, noise reduction, and signal enhancement on the brain electrical signals. The data analysis processing shared platform 21 can perform real-time monitoring, coding and decoding, data sharing, and the like on the brain electrical signals. The integration of the automated miniaturized brain organoid culture system and the brain electrical signal input and output system not only stimulates and trains the brain organoid during the brain organoid culture process, but also collects, processes, and analyzes the electrical signals generated by the brain organoid in real time, and realizes data sharing through the shared platform, builds a remote collaborative biological computer, and develops personalized brain-like intelligence.

[0037] In summary, the brain organoid long-term culture device has the following technical effects:

[0038] The brain organoid long-term culture device integrates brain organoid culture in a microfluidic chip, realizes automatic culture of brain organoids, can accurately control the culture microenvironment, reduces waste of culture medium, saves manpower and resources, and is more conducive to realizing standardized and batch culture of brain organoids, and efficiently assists brain science research.

[0039] The brain organoid long-term culture device adopts microelectrode array technology and microscopic imaging technology, and can obtain brain electrical signals and monitor the growth state of brain organoids in real time during the culture process.

[0040] The electroencephalogram signal processor can perform preliminary processing such as filtering, noise reduction, and signal enhancement on the electroencephalogram signals. The data analysis and processing sharing platform can perform real-time monitoring, encoding and decoding, and data sharing of the electroencephalogram signals.

[0041] Although the present application is specifically shown and described in connection with preferred embodiments, those skilled in the art will appreciate that various modifications in form and detail can be made without departing from the spirit and scope of the application as defined by the appended claims.

Claims

1. A long-term brain organoid culture device, characterized in that, include: Incubator, brain organoid chip, microelectrode array, microscopic imaging module and oscillation device; The incubator is used to provide a culture environment for brain organoids in brain organoid chips; The brain organoid chip comprises: a light-transmitting glass layer on top and a base layer on the bottom; a container is formed in the center of the base layer, the upper edge of the container is connected to the outside of the chip through an inlet pipe, and the bottom of the container is connected to the outside of the chip through an outlet pipe; the space of the container is divided into two parts by a semi-permeable membrane, the bottom of the container is used to store culture medium, and the area above the semi-permeable membrane is used to place a brain organoid matrix gel mixture, the semi-permeable membrane is used for the exchange of nutrients between the brain organoid matrix gel mixture and the culture medium; The microelectrode array is closely covered above the brain organoid matrix gel mixture and is led out through the EEG signal transmission line for collecting EEG signals; The microscopic imaging module is used to acquire images of brain organoids on the brain organoid chip.

2. The brain organoid long-term culture device as described in claim 1, characterized in that: The container is a cylindrical container, and the semi-permeable membrane covers the container and sinks to form a hemispherical space.

3. The brain organoid long-term culture device as described in claim 1, characterized in that: The brain organoid chip has a detachable structure, with the four corners of its upper and lower layers fixed with screws.

4. The brain organoid long-term culture device as described in claim 1, characterized in that: The microelectrode array adopts a flexible, rhomboid mesh structure.

5. The brain organoid long-term culture device as described in claim 1, characterized in that: The microscopic imaging module uses a macro camera with a focal length of 50mm to 100mm.

6. The brain organoid long-term culture device as described in claim 1, characterized in that: The incubator includes a chamber, a control module, and a sterilization module, a temperature module, a humidity module, and a carbon dioxide concentration monitoring module electrically connected to the control module.

7. The brain organoid long-term culture device as described in claim 6, characterized in that: The sterilization module is an ultraviolet lamp.

8. The brain organoid long-term culture device as described in claim 6, characterized in that: The incubator is also equipped with an oscillation device for oscillating the brain organoid chip.

9. The brain organoid long-term culture device as described in claim 8, characterized in that: The oscillation device is an oscillation table or a vibrating arm.

10. A long-term brain organoid culture system, characterized in that: The device includes a culture medium, a waste liquid storage device, an EEG signal processor, and a long-term brain organoid culture device as described in any one of claims 1-9; the culture medium is connected to the inlet pipe of the long-term brain organoid culture device for providing culture medium, and the waste liquid storage device is connected to the outlet pipe of the long-term brain organoid culture device for collecting waste liquid; both the inlet pipe and the outlet pipe are equipped with microfluidic control valves; and the microelectrode array is connected to the EEG signal processor via an EEG signal transmission line.

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