Organ-like culture chamber

Abstract

The organ-like culture chamber comprises a chamber body and a chamber end cover which are connected with each other, the chamber body is provided with a cylindrical containing cavity with an opening in one side, a fluid inlet and a fluid outlet, the fluid inlet and the fluid outlet are communicated with the containing cavity, and the chamber end cover is detachably connected to the opening end of the chamber body in a sealed mode. The cavity end cover is of a square structure, the open end of the cavity body is provided with an end cover groove matched with the cavity end cover, and the cavity end cover is connected into the end cover groove through a connecting piece. And the sealing ring is arranged between the chamber end cover and the chamber main body so as to seal the open end of the chamber main body. The fluid inlet and the fluid outlet are respectively provided with a connector.

Description

Technical Field

[0001] The present application belongs to the field of biomedical engineering, and specifically relates to an organoid culture chamber. Background Art

[0002] With the continuous advancement of biomedical research, organoid culture technology has gradually become an important tool for simulating and studying the growth environment and function of organoids. However, traditional organoid culture methods have certain limitations, such as unstable culture environment, organoids easily settle or adhere to the wall of the culture container, and it is difficult to maintain a suspended state. In addition, the gas exchange efficiency is low and cannot meet the metabolic needs of organoids, resulting in unsatisfactory culture effects. These problems seriously affect the culture efficiency and quality of organoids, limiting their widespread application in biomedical research and applications. Existing organoid culture techniques usually rely on flat culture dishes or multi-well plates, which have certain limitations in organoid growth and proliferation.

[0003] In order to improve the efficiency and quality of organoid culture, a new organoid culture chamber is needed. Summary of the invention

[0004] Purpose: In view of at least one of the above technical problems, the present application provides a more efficient, stable and flexible organoid culture chamber by combining microgravity suspension culture, multi-channel liquid supply and efficient gas exchange technologies.

[0005] Technical solution: To solve the above technical problems, the technical solution adopted in this application is: According to a first aspect of the present application, there is provided an organoid culture chamber, comprising a chamber body and a chamber end cover connected to each other, the chamber body having a cylindrical accommodating cavity with an opening on one side and a fluid inlet and a fluid outlet connected to the accommodating cavity, and the chamber end cover being detachably sealed and connected to the open end of the chamber body.

[0006] In some embodiments, the chamber end cover is a square structure, the open end of the chamber body is provided with an end cover groove matching with the chamber end cover, and the chamber end cover is connected to the end cover groove through a connector.

[0007] In some embodiments, threaded countersunk holes are provided at the four corners of the end cover groove, and through holes are opened at corresponding positions of the chamber end cover, which are used to pass set screws through the through holes and then be threadedly connected to the threaded countersunk holes, so that the chamber end cover can be detachably connected to the chamber body.

[0008] In some embodiments, an outer side of the chamber end cap is flush with a corresponding side on the chamber body.

[0009] In some embodiments, the organoid culture chamber further comprises a sealing ring, which is disposed between the chamber end cover and the chamber body to seal the open end of the chamber body.

[0010] In some embodiments, the fluid inlet and the fluid outlet are respectively provided with connectors, and in this embodiment, the connectors are Luer connectors.

[0011] In some embodiments, the fluid inlet and the fluid outlet are arranged perpendicular to each other.

[0012] In some embodiments, the chamber body is made of transparent material to facilitate observation of the organoid culture conditions in the containing cavity.

[0013] In some embodiments, the chamber body is provided with a bead groove for cooperating with a bead on a culture chamber support, so as to fix the organoid culture chamber on the culture chamber support through the bead and the bead groove.

[0014] Beneficial effects: The organoid culture chamber provided in the present application is suitable for independent and joint culture of multiple organoids, can realize suspension culture of organoids under microgravity conditions, and has functions such as gas exchange and real-time monitoring. It has the following advantages: combined with the microgravity suspension culture system, the culture medium delivery system, the gas exchange system and the real-time monitoring system, the chamber body is made of polycarbonate material, which has excellent transparency and chemical stability; specifically, the chamber can realize the following functions: 1. Microgravity suspension culture: through the microgravity rotation perfusion system, the organoids are evenly distributed in the culture medium and kept in a suspended state to prevent sedimentation or adhesion. Control the rotation speed of the stepper motor to significantly improve the stability of the organoid suspension culture. It is composed of a micropump and a microfluidic channel. The micropump is piezoelectrically driven and can accurately control the flow rate; the culture medium circulates during the culture process through a peristaltic pump to maintain the freshness and nutrient supply of the culture medium. 2. Efficient gas exchange: It is equipped with an oxygenator component and adopts an efficient gas exchange design to achieve efficient exchange of carbon dioxide and oxygen, achieve blood oxygen balance, and meet the metabolic needs of organoids. Utilize breathable silicone rubber tubing and adjust the silicone rubber tubing process to make the rubber tubing have good air permeability and barrier properties. 3. Real-time monitoring system: Install a camera at the front of the chamber for real-time monitoring, which is convenient for observing the growth and suspension state of organoids, and automatically adjusts system parameters through image recognition technology to maintain optimal culture conditions. The monitoring system can automatically adjust system parameters to ensure that organoids grow and proliferate under optimal conditions. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is a schematic diagram of the structure of the organoid culture chamber according to an embodiment of the present application; Figure 2This is a schematic diagram of the assembly of components of the organoid culture chamber in an embodiment of the present application; Figure 3 This is a cross-sectional schematic diagram of an organoid culture chamber in an embodiment of the present application; Figure 4 This is a schematic diagram of the working principle of the organoid culture chamber system in the embodiment of the present application; Figure 5 This is a schematic diagram of the structure of a culture chamber support for installing an organoid culture chamber in an embodiment of the present application; Figure 6 This is a schematic diagram of the suspension test of the organoid culture chamber in the embodiment of the present application; In the figure: camera 2, nutrient solution filling 3, peristaltic pump 4, oxygenator 5; culture chamber 6, chamber body 601, end cover groove 601a, chamber end cover 602, accommodating cavity 603, fluid inlet 604 and fluid outlet 605, set screw 606, sealing ring 607, joint 608, ball groove 609; culture chamber support 7, culture chamber placement groove 701, dovetail structure 702, ball 703. DETAILED DESCRIPTION

[0016] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present application and its application or use. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0017] Unless otherwise specifically stated, the relative arrangement, numerical expressions and numerical values ​​of the parts and steps set forth in these embodiments do not limit the scope of the application. Meanwhile, it should be understood that, for ease of description, the sizes of the various parts shown in the accompanying drawings are not drawn according to actual proportional relationships. The technology, methods and equipment known to those of ordinary skill in the relevant art may not be discussed in detail, but in appropriate cases, the technology, methods and equipment should be considered as a part of the specification. In all examples shown and discussed here, any specific value should be interpreted as being merely exemplary, rather than as a limitation. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar reference numerals and letters represent similar items in the following drawings, and therefore, once a certain item is defined in an accompanying drawing, it does not need to be further discussed in subsequent drawings.

[0018] In the description of the present application, it should be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "inside", "outside", etc., indicating the orientation or position relationship, are based on the orientation or position relationship shown in the drawings, and are only used to explain the relative position relationship, movement, etc. between the components in a certain posture. If the specific posture changes, the directional indication will also change accordingly. It is only for the convenience of describing the present application and simplifying the description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation to the present application.

[0019] In addition, the terms "first", "second", etc. are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Thus, a feature defined as "first", "second", etc. may explicitly or implicitly include one or more of the feature. In the description of this application, unless otherwise specified, "plurality" means two or more.

[0020] In the description of this application, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in this application can be understood by specific circumstances.

[0021] Example 1: Figures 1 to 6 As shown, this embodiment provides an organoid culture chamber 6, which includes a chamber body 601 and a chamber end cover 602 connected to each other, wherein the chamber body 601 has a cylindrical accommodating cavity 603 with an opening on one side, and a fluid inlet 604 and a fluid outlet 605 connected to the accommodating cavity 603, and the chamber end cover 602 is detachably sealed and connected to the open end of the chamber body 601.

[0022] In this embodiment, the accommodating cavity 603 is designed as a cylindrical structure with a smooth interior and no dead corners, which is convenient for cleaning and disinfection and can effectively prevent the organoids from adhering to the cavity wall.

[0023] In some embodiments, Figure 1 , Figure 2As shown, the chamber end cover 602 is a square structure, the open end of the chamber body 601 is provided with an end cover groove 601a that matches the chamber end cover 602, and the chamber end cover 602 is connected to the end cover groove 601a through a connector. In this embodiment, the end cover groove 601a is a square structure.

[0024] In some embodiments, Figure 2 As shown, threaded countersunk holes are arranged at the four corners of the end cover groove 601a, and through holes are opened at corresponding positions of the chamber end cover 602, which are used to pass the set screws 606 through the through holes and then be threadedly connected in the threaded countersunk holes, so that the chamber end cover 602 can be detachably connected to the chamber body 601.

[0025] In some embodiments, Figure 1 , Figure 5 As shown, the outer side of the chamber end cover 602 is flush with the corresponding side of the chamber body 601. This facilitates fixing the organoid culture chamber in the culture chamber placement slot on the culture chamber support.

[0026] Furthermore, in some embodiments, Figure 2 As shown, the culture chamber further includes a sealing ring 607, and the sealing ring 607 is arranged between the chamber end cover 602 and the chamber body 601 to seal the open end of the chamber body.

[0027] It should be noted that the sealing ring is an important component to ensure the stability of the environment in the chamber. It is made of food-grade silicone material, has excellent elasticity, sealing performance and high temperature resistance, and can withstand chemical corrosion from the culture medium. In this embodiment, the sealing ring is designed as an O-ring, which is embedded in the joint between the chamber end cover and the chamber body to ensure that it can fit tightly between the chamber body and the chamber end cover after installation to prevent liquid leakage. The sealing ring is embedded between the contact surface of the chamber body and the chamber end cover, and the sealing effect is achieved through the clamping force. The design of the sealing ring also takes into account the characteristics of easy replacement and cleaning, which is convenient for multiple use.

[0028] In some embodiments, Figure 1 , Figure 2As shown, the fluid inlet 604 and the fluid outlet 605 are respectively provided with a connector 608. The connector in this embodiment adopts a Luer connector. The Luer connector is a standardized micro-seepage-free connector, which is connected by a male Luer connector and a matching female Luer connector part. The Luer connector is used to connect the culture medium delivery pipeline, the gas exchange system and the real-time monitoring equipment. These interfaces adopt standardized interface sizes to ensure the sealing and stability of the connection. The connector in this application adopts a standardized design, which is convenient for connection with various pipelines and equipment. The connector is used to connect the culture medium delivery system and the gas exchange system, and corrosion-resistant plastic materials are used to ensure that no corrosion or leakage occurs during long-term use. The connector is designed as a standardized threaded interface, which is convenient for quick connection and disassembly with various pipelines and equipment. The joint adopts a double sealing design, including a built-in O-ring and an external sealing gasket, to ensure the sealing and stability of the connection. The design of the connector also takes into account the needs of rapid disassembly and assembly, which is convenient for maintenance and cleaning of the system.

[0029] In some embodiments, Figure 2 , Figure 3 , Figure 6 As shown, the fluid inlet 604 and the fluid outlet 605 are arranged perpendicular to each other.

[0030] In some embodiments, the chamber body 601 and the chamber end cover 602 are made of transparent materials to facilitate observation of the organoid culture conditions in the chamber. Figure 4 As shown, the organoid culture conditions in the accommodating chamber are collected by camera 2. Furthermore, in this embodiment, the chamber body is designed to be made of high-strength, corrosion-resistant transparent materials to ensure good visibility and chemical stability, which is convenient for observing the growth of the organoids in the accommodating chamber. For example, polycarbonate material can be selected, which is not only transparent, but also has excellent impact resistance and compressive strength, and can withstand the mechanical stress generated during efficient operation.

[0031] The chamber end cap is an important part of the closed chamber. It is made of the same transparent material as the chamber body, and also uses polycarbonate material to maintain consistency with the chamber body. The chamber end cap is designed to be a detachable structure, which is convenient for operators to add or remove samples during the experiment. An observation window and a connector interface are reserved on the chamber body to facilitate the connection of the real-time monitoring system and the gas exchange system. The chamber end cap is fastened to the chamber body by screws to ensure that the chamber end cap can be stably fixed after installation, and a good sealing effect is achieved through the sealing ring.

[0032] In some embodiments, Figure 1 , Figure 3 , Figure 5As shown, the chamber body 6 is provided with a ball groove 609 for cooperating with the ball 703 on the culture chamber support 7, so as to fix the culture chamber on the culture chamber support through the ball and the ball groove 609.

[0033] In this embodiment, Figure 5 As shown, the culture chamber support 7 is divided into eight parallel culture chamber placement slots 701 by a partition in the length direction; corresponding positions on both sides in the width direction are provided with shifting beads 703. No dovetail structures 702 are provided at both ends of the culture chamber support 7 in the length direction.

[0034] In some embodiments, Figure 6 Shown is a schematic diagram of the organoid culture chamber suspension test.

[0035] The organoid culture chamber occupies the main part of the entire culture system and has good sealing properties. It is used for suspended culture of organoid matrix. The platform can be driven by a stepper motor and a microgravity environment can be achieved by adjusting the rotation speed. The culture fluid is delivered to the main body of the chamber through a micropump and circulated through a peristaltic pump. The oxygenator assembly and the breathable silicone rubber tube ensure efficient gas exchange. The real-time monitoring system uses a camera and image recognition technology to achieve real-time monitoring and automatic adjustment of the organoid culture status.

[0036] Working principle: 1. The microgravity suspension culture system creates a microgravity environment in the culture chamber by means of rotary perfusion. The chamber body is mounted on a rotatable platform driven by a stepper motor. By controlling the rotation speed of the stepper motor, the rotation speed of the chamber can be adjusted to adjust the intensity of the microgravity environment. When the chamber body rotates, the culture fluid and organoids form a dynamic balance under the combined action of centrifugal force and gravity, and the organoids remain suspended in the culture fluid to prevent the organoids from settling or adhering to the chamber wall. The rotary perfusion system ensures that the organoids are evenly distributed in the culture fluid, which helps the organoids to evenly receive nutrients and gas exchange. 2. The culture fluid delivery system maintains the freshness and nutrient supply of the culture fluid by precisely controlling the flow rate and circulation of the culture fluid. The micropump is piezoelectrically driven and can precisely control the flow rate. The culture fluid is delivered to the chamber body through the micropump to ensure a stable supply of the culture fluid. The culture fluid circulates during the culture process through a peristaltic pump to prevent the accumulation of waste in the culture fluid and ensure that the organoids grow in a constant environment. By adjusting the flow rate of the micropump, the supply of culture fluid can be precisely controlled according to the needs of the organoids. 3. The efficient gas exchange system realizes efficient exchange of gas in the culture fluid through the permeable material and oxygenator assembly to meet the metabolic needs of the organoids. The oxygenator assembly is installed on the upper part of the culture chamber, and the gas exchange is realized through the permeable silicone rubber tube. The silicone rubber tube has good permeability and barrier properties, and can effectively exchange carbon dioxide and oxygen. By adjusting the area and gas flow of the silicone rubber tube, the gas balance in the culture fluid is ensured to achieve the optimal gas environment required by the organoid. The efficient gas exchange system can also effectively discharge the waste gas generated during the metabolism of the organoid to prevent the accumulation of waste gas from affecting the growth of the organoid. 4. The real-time monitoring system monitors the growth and suspension state of the organoid in real time through the camera installed at the front of the chamber. The camera is installed at the front of the chamber, and the growth and suspension state of the organoid are monitored in real time through high-definition image transmission technology. The monitoring system is equipped with image recognition technology, which can automatically identify the distribution and state of the organoid and detect abnormal conditions. According to the monitored organoid status, the system can automatically adjust the rotation speed, culture fluid flow rate and gas exchange parameters to ensure that the organoids grow and proliferate under optimal conditions. All systems are integrated and controlled by a central controller, which dynamically adjusts the working parameters of each system based on the data provided by the real-time monitoring system to ensure a stable and optimal organoid culture environment.

[0037] Application Example: Use of Organoid Culture Chamber Step S1: Preparation 1. Cleaning and disinfection: The chamber is sterilized with high temperature and high pressure before use, and then the sterile operating table is used to thoroughly soak and clean the organoid culture chamber with ultrapure water, and finally irradiated with ultraviolet light for two hours to ensure that there is no pollution source in the chamber. 2. Assemble the chamber: Assemble the chamber body, microgravity suspension culture system, culture fluid delivery system, gas exchange system and real-time monitoring system according to the design drawings, and ensure that all components are tightly connected and leak-free.

[0038] Step S2: Configuration of culture medium delivery system 1. Transfer the decellularized matrix containing stem cells: transfer the decellularized matrix that has been attached to stem cells for 24 hours into the chamber, seal the chamber, and check the sealing of the joints. 2. Add culture medium: after the test is correct, add the prepared culture medium into the culture liquid bottle, and prevent a large number of bubbles from appearing on the liquid surface. 3. Infuse nutrient solution: after the bubbles on the liquid surface of the culture medium in the nutrient solution bottle completely disappear, turn on the peristaltic pump, let the culture medium slowly enter the chamber through the silicone tube, slowly fill the entire circuit, and finally flow back into the bottle from the outlet.

[0039] Step S3: Configuration of an efficient gas exchange system 1. Install the oxygenator assembly: Fix the oxygenator assembly on the upper part of the chamber body and connect the breathable silicone rubber tube. 2. Adjust the gas flow: Set the gas flow requirement of the oxygenator and replace the breathable silicone tube of the corresponding model to ensure the optimal exchange of carbon dioxide and oxygen. 3. Check the air tightness: Make sure that all gas exchange pipes and interfaces are leak-free to prevent outside air from entering the chamber.

[0040] Step S4: Installation and debugging of real-time monitoring system 1. Install the camera: Fix the high-definition camera at the front of the chamber to ensure that the camera's field of view covers the entire culture area. 2. Adjust the camera's focal length and angle: Ensure that the camera can clearly capture the growth and suspension status of the organoids. 3. Connect the image recognition system: Connect the camera to the image recognition system, use wireless transmission technology to provide real-time feedback on the position of the matrix in the chamber, and use the image recognition system to determine the matrix suspension status for flow control.

[0041] Step S5: Start the system and culture organoids 1. Start the culture medium delivery system: Start the micro pump through the central controller to start circulating the culture medium to ensure that the cells get enough nutrients. 2. Start the efficient gas exchange system: Start the oxygenator component through the central controller to perform gas exchange and maintain the gas balance in the culture medium.

[0042] Step S6: Real-time adjustment and maintenance 1. Start the real-time monitoring system: Turn on the camera and image recognition system to monitor the growth and suspension status of the organoids in real time. Through the image recognition system, analyze the growth of the organoids in real time and detect the distribution and status of the organoids. 2. Automatically adjust parameters: According to the monitoring data, the central controller automatically adjusts the rotation speed, culture fluid flow rate and gas exchange parameters to ensure that the organoids grow under optimal conditions. 3. Regularly replace the culture medium: According to the organoid culture cycle, regularly replace the fresh culture medium to prevent waste accumulation from affecting the growth of the organoids.

[0043] Step S7: End of culture and harvesting of organoids 1. Stop system operation: When the organoid reaches the expected growth state or the culture cycle ends, the central controller stops the operation of each system in turn. 2. Harvest organoids: Under sterile conditions, the culture fluid in the chamber body is transferred to the collection container through a peristaltic pump for subsequent processing and analysis. 3. Cleaning and disinfection: After the organoid harvest is completed, the organoid culture chamber is thoroughly cleaned and disinfected to prepare for the next culture cycle.

[0044] In summary, the organoid culture chamber provided in this application can achieve the following functions: 1. Microgravity suspension culture: Through the microgravity rotary perfusion system, the organoids are evenly distributed in the culture medium and kept in a suspended state to prevent sedimentation or adhesion. Control the rotation speed of the stepper motor to significantly improve the stability of the organoid suspension culture. It is composed of a micropump and a microfluidic channel. The micropump is piezoelectrically driven and can accurately control the flow rate; the culture medium circulates during the culture process through a peristaltic pump to maintain the freshness and nutrient supply of the culture medium. 2. Efficient gas exchange: It is equipped with an oxygenator component and adopts an efficient gas exchange design to achieve efficient exchange of carbon dioxide and oxygen, achieve blood oxygen balance, and meet the metabolic needs of the organoid. Using a breathable silicone rubber tube, by adjusting the silicone rubber tube process, the rubber tube has good permeability and barrier properties. 3. Real-time monitoring system: A camera is installed at the front of the chamber for real-time monitoring, which is convenient for observing the growth and suspension state of the organoid, and automatically adjusts the system parameters through image recognition technology to maintain the optimal culture conditions. The monitoring system automatically adjusts system parameters to ensure that organoids grow and proliferate under optimal conditions.

[0045] The technical means disclosed in the present application are not limited to the technical means disclosed in the above implementation mode, but also include technical solutions composed of any combination of the above technical features. The present application has been disclosed in the above preferred embodiments, but it is not used to limit the present application. Any technical solution obtained by equivalent replacement or equivalent transformation falls within the protection scope of the present application.

Claims

1. An organoid culture chamber, It is characterized in that The organoid culture chamber includes a chamber body and a chamber end cover connected to each other, wherein the chamber body has a cylindrical accommodating cavity with an opening on one side and a fluid inlet and a fluid outlet connected to the accommodating cavity, and the chamber end cover is detachably sealed and connected to the open end of the chamber body.

2. The organoid culture chamber according to claim 1, It is characterized in that The chamber end cover is a square structure, and the opening end of the chamber body is provided with an end cover groove matching with the chamber end cover, and the chamber end cover is connected to the end cover groove through a connecting piece.

3. The organoid culture chamber according to claim 2, It is characterized in that Threaded countersunk holes are arranged at the four corners of the end cover groove, and through holes are opened at corresponding positions of the chamber end cover, which are used for threading the chamber end cover into the threaded countersunk holes after the set screws pass through the through holes, so as to detachably connect the chamber end cover to the chamber body.

4. The organoid culture chamber according to claim 2, It is characterized in that The outer side of the chamber end cover is flush with the corresponding side on the chamber body.

5. The organoid culture chamber according to claim 1, It is characterized in that A sealing ring is also included, and the sealing ring is arranged between the chamber end cover and the chamber body to seal the open end of the chamber body.

6. The organoid culture chamber according to claim 1, It is characterized in that The fluid inlet and the fluid outlet are respectively provided with joints.

7. The organoid culture chamber according to claim 6, It is characterized in that The connector is a Luer connector.

8. The organoid culture chamber according to any one of claims 1 to 7, It is characterized in that The fluid inlet and the fluid outlet are arranged perpendicular to each other.

9. The organoid culture chamber according to any one of claims 1 to 7, It is characterized in that The chamber body and the chamber end cover are made of transparent materials, which facilitates observation of the organoid culture conditions in the accommodating cavity.

10. The organoid culture chamber according to any one of claims 1 to 7, It is characterized in that The chamber body is provided with a bead groove for cooperating with a bead on the culture chamber support, so as to fix the organoid culture chamber on the culture chamber support through the bead and the bead groove.

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