Multi-cavity somatic cell culture device for co-culture of macrophages
By using a ring distributor and a biofiltration membrane structure in the macrophage co-culture device, the problem of uneven distribution of signal molecules was solved, and uniform distribution of culture medium and stable growth of production cells were achieved.
Patent Information
- Application Number
- CN202521695989.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-11
- Publication Date
- 2025-09-12
- Estimated Expiration
- 2035-08-11
AI Technical Summary
In the prior art, single-point injection of culture medium during macrophage co-culture results in uneven distribution of signal molecules, affecting the consistency of production cell growth and low efficiency of signal molecule utilization.
A multi-chamber cell culture device is used. An annular distributor and a biological filtration membrane are set in the second culture chamber. The port of the connecting pipeline in the second culture chamber is designed as an annular distributor. Several through holes are opened on the inner wall and anti-disturbance wings are equipped to ensure uniform distribution of culture medium and cell isolation.
The uniform release of cytokines in the culture medium is achieved, the transmission efficiency of signal molecules and the growth consistency of production cells are improved, and the problem of unstable growth caused by uneven cytokine concentration is avoided.
Smart Images
Figure CN223329327U_ABST
Abstract
Description
Technical Field
[0001] The utility model belongs to the technical field of cell culture, and in particular relates to a multi-chamber cell culture device for co-culturing macrophages. Background Art
[0002] Macrophages are a key cell type in the immune system. They not only engulf foreign pathogens and cellular debris, but also initiate and regulate complex immune responses by secreting a variety of signaling molecules called cytokines. Due to their central role in health and disease, macrophages have become an important subject of biomedical research.
[0003] In a typical application scenario, macrophages themselves are the "producer cells" that need to be produced or studied in large quantities. In order to stimulate their proliferation or differentiate into subtypes with specific functions, researchers will use another type of engineered cell that can continuously secrete specific cytokines as an auxiliary.
[0004] To ensure that the cytokines secreted by the engineered cells effectively act on the production cells (such as macrophages) during the co-culture process while avoiding physical contact between the two cells, which could lead to impure final cell products, existing technologies typically employ a scheme that physically isolates the two culture chambers while allowing the culture medium to flow through them. For example, a connecting pipe with a built-in biofiltration membrane connects the engineered cell chamber to the production cell chamber. This structure effectively isolates the two cell types, while allowing the cytokines secreted by the engineered cells to passively diffuse through the biofiltration membrane and into the production cell chamber, achieving non-contact stimulation culture.
[0005] However, the connecting line at the production cell chamber typically has a simple orifice, meaning that the cytokine-rich culture medium enters the entire culture system from a single point. This single-point injection method results in extremely uneven distribution of signaling molecules, with concentrations being too high near the orifice and too low farther away. This results in significant variations in the intensity of stimulation received by the cell populations (specifically, macrophages) within the chamber. This uneven culture microenvironment not only affects the consistency of production cell growth but also reduces the overall utilization efficiency of signaling molecules. Utility Model Content
[0006] In order to solve the above-mentioned problems in the prior art, the present invention provides a multi-chamber cell culture device for co-culturing macrophages.
[0007] In order to achieve the above purpose, the technical solution adopted by the utility model is:
[0008] Provided is a multi-chamber cell culture device for macrophage co-culture, comprising:
[0009] a first culture chamber configured for seeding engineered cells;
[0010] a second culture chamber configured for seeding producer cells;
[0011] a communication pipe, wherein both ends of the communication pipe are respectively connected to the first culture chamber and the second culture chamber to establish a fluid passage;
[0012] Wherein, the port of the communication pipeline in the second culture chamber is configured as a ring distributor;
[0013] The annular distributor is arranged along the inner circumferential wall of the second culture chamber and maintains a preset distance from the bottom surface of the second culture chamber;
[0014] A plurality of through holes are provided on the inner side wall of the annular distributor;
[0015] Furthermore, a biological filtration membrane for isolating cells is provided inside the communicating pipe.
[0016] Preferably, the axial directions of the plurality of through holes formed on the inner side wall of the annular distributor are arranged to be horizontally directed toward the center of the second culture chamber, or to be inclined downward at a preset angle to the horizontal plane.
[0017] Preferably, it includes:
[0018] an anti-disturbance wing, arranged below the annular distributor;
[0019] Wherein, the anti-disturbance wing is configured to block the medium flowing out of the through hole.
[0020] Preferably, the anti-disturbance wing portion is an umbrella-shaped shield with a downwardly inclined upper surface in a conical or arc shape.
[0021] Preferably, the biological filtration membrane is positioned in a filtration module;
[0022] Furthermore, the filter module has a first port and a second port;
[0023] The first port and the second port are respectively detachably connected to the communication pipe.
[0024] Preferably, the filtering module includes:
[0025] A filter membrane box composed of an upper shell and a lower shell that can be detachably matched with each other;
[0026] at least one porous support plate disposed inside the membrane cartridge and configured to support the biological filtration membrane;
[0027] and at least one elastic sealing ring disposed in the filter membrane box and configured to compress and seal the edge of the biological filter membrane.
[0028] Preferably, a spherical buffer cavity is provided in series in the communication pipeline, and the spherical buffer cavity is located between the first culture chamber and the filtration module.
[0029] Preferably, the upper portion of the spherical buffer cavity has a gas phase space for collecting gas, and a hydrophobic breathable membrane is provided on the top of the gas phase space to allow the collected gas to discharge from the buffer cavity.
[0030] Preferably, the connecting pipeline has:
[0031] The valve or pipe clamp is configured to cut off or open the connection state of the connecting pipeline.
[0032] Preferably, the second culture chamber has:
[0033] The sampling port is sealed by a detachable sealing cover.
[0034] The present invention provides a multi-chamber cell culture device for macrophage co-culture. The beneficial effects of the present invention are as follows:
[0035] Driven by concentration differences or slight liquid level differences, cytokine-rich culture medium flows through connecting pipes, passes through a biological filtration membrane, and reaches an annular distributor. Through several through-holes on the inner wall of the annular distributor, it is gently and evenly released into the entire culture environment of the second culture chamber, thereby efficiently and uniformly stimulating the growth or functional polarization of producer cells. This annular distributor structure effectively solves the problems of uneven mixing of signal molecules and low stimulation efficiency caused by single-point injection in existing technologies. BRIEF DESCRIPTION OF THE DRAWINGS
[0036] Figure 1 This is a cross-sectional view of the multi-chamber cell culture device for macrophage co-culture proposed in the present invention;
[0037] Figure 2 for Figure 1 A local enlarged schematic diagram at point A;
[0038] Figure 3 This is a cross-sectional view of the filtration module in the multi-chamber cell culture device for macrophage co-culture proposed in the present invention.
[0039] Description of reference numerals:
[0040] 1. First culture chamber; 2. Second culture chamber; 3. Connecting pipe; 4. Ring distributor; 5. Biological filter membrane; 6. Anti-disturbance wing; 7. Filtration module; 701. First port; 702. Second port; 703. Filter membrane box; 704. Porous support plate; 705. Elastic sealing ring; 8. Spherical buffer chamber; 801. Gas phase space; 802. Hydrophobic breathable membrane; 9. Sampling port. DETAILED DESCRIPTION
[0041] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.
[0042] See also Figure 1-Figure 3 As shown, the specific embodiments provided by the present invention are as follows:
[0043] like Figure 1 As shown, an embodiment of the present invention provides a multi-chamber cell culture device for co-culture of macrophages, which mainly includes a first culture chamber 1, a second culture chamber 2 and a connecting pipe 3.
[0044] The first culture chamber 1, as an independent culture unit, has an interior space configured for seeding and culturing a first type of cell, preferably engineered cells in this embodiment. The second culture chamber 2, as another independent culture unit, has an interior space configured for seeding and culturing a second type of cell, preferably producer cells in this embodiment, such as macrophages requiring stimulated culture. Each chamber is equipped with a sealing cover for aseptic operation and gas exchange (e.g., provided with a gas supply line).
[0045] The two ends of the communication pipe 3 are connected to the outlet of the first culture chamber 1 and the inlet of the second culture chamber 2 respectively through sterile interfaces, thereby establishing a closed fluid passage between the two chambers.
[0046] The end of the connecting pipe 3 in the second culture chamber 2 is an annular distributor 4. The annular distributor 4 is arranged along the inner circumferential wall of the second culture chamber 2 and, through its own structure or a supporting structure, maintains a preset distance from the bottom surface of the second culture chamber 2, thereby avoiding direct contact with the cells cultured at the bottom.
[0047] The inner wall of the annular distributor 4, i.e., the side facing the center of the second culture chamber 2, is uniformly provided with a plurality of through-holes. These through-holes serve as the final outlet for the culture medium, allowing the liquid flowing in from the connecting pipe 3 to be released simultaneously and evenly from all sides into the culture environment of the second culture chamber 2.
[0048] To achieve physical isolation between the two cell types, a biofiltration membrane 5 is installed inside the connecting pipe 3 or inside the annular distributor 4. The pore size of the biofiltration membrane 5 is small enough to effectively block the passage of any cells, but allows the culture medium and small molecules such as cytokines dissolved therein to freely penetrate.
[0049] The working principle of the present invention is as follows: first, engineered cells are inoculated and cultured in the first culture chamber 1, and production cells (such as macrophages) are inoculated and cultured in the second culture chamber 2. After the culture starts, the engineered cells in the first culture chamber 1 will secrete cytokines into their culture medium. Driven by concentration differences or slight liquid level differences, these cytokine-rich culture media pass through the connecting pipe 3, are filtered through the biological filtration membrane 5, and then reach the annular distributor 4. Subsequently, the culture medium is gently and evenly released into the entire culture environment of the second culture chamber 2 through a number of through holes on the inner wall of the annular distributor 4, thereby efficiently and uniformly stimulating the growth or functional polarization of the production cells. The present invention effectively solves the problems of uneven mixing of signal molecules and low stimulation efficiency caused by single-point injection in the prior art through the structure of the annular distributor 4.
[0050] In a preferred embodiment, the axial directions of the plurality of through holes formed on the inner wall of the annular distributor 4 are set to be horizontally directed toward the center of the second culture chamber 2 or inclined downward at a preset angle to the horizontal plane.
[0051] This structure actively organizes and guides fluid movement within the second culture chamber 2, achieving more efficient mixing. When the through-holes are arranged horizontally, liquid flowing from the periphery toward the center creates a gentle, integrated horizontal circulation in the upper-middle region of the culture medium. When the through-holes are arranged downwardly, they promote a more three-dimensional vertical convection of the culture medium, flowing from the upper side walls to the center of the bottom, and then circulating upward.
[0052] In this flow field mode, the newly flowing cytokine-rich culture medium can be efficiently and evenly mixed with the original culture medium in the chamber without generating direct fluid shear force impact on the cells growing on the bottom wall, thereby further improving the transmission efficiency and effect of the signal molecules and ensuring that all production cells in the chamber can receive uniform and stable signal stimulation.
[0053] In a preferred embodiment, the multi-chamber cell culture device for macrophage co-culture further includes an anti-disturbance wing 6, which is disposed below the annular distributor 4. The anti-disturbance wing 6 is configured to block the medium flowing out of the through-holes on the inner wall of the annular distributor 4.
[0054] Specifically, the anti-disturbance wing 6 is an annular or sheet-like structure, which can be connected to the bottom of the annular distributor 4 through a fixing part, or directly connected to the inner wall of the second culture chamber 2 through a supporting structure. The main function of the anti-disturbance wing 6 is to act as a physical barrier to intercept the culture medium fluid ejected from the through hole to prevent it from directly impacting the cell layer at the bottom of the second culture chamber 2. This is suitable for protecting production cells (such as macrophages) that grow adherently to the wall and avoid damage due to local high flow rate or mechanical disturbance. By introducing the anti-disturbance wing 6, the utility model can further optimize the stability of the microenvironment in the second culture chamber 2 on the basis of maintaining uniform distribution of the culture medium, ensuring that the production cells can grow and differentiate efficiently without interference from fluid shear force.
[0055] In a preferred embodiment, the anti-interference wing portion 6 is an umbrella-shaped shield with a downwardly inclined upper surface in a conical or arc shape.
[0056] Specifically, as culture medium flows out of the through-hole, it first contacts the inclined surface of the umbrella-shaped shield and then slowly slides along this surface toward the bottom of the second culture chamber 2. This umbrella-shaped shield extends the flow path of the culture medium and significantly reduces the impact velocity of the fluid, thereby achieving more gentle medium injection and avoiding direct disturbance of adherent cells. Furthermore, the user can adjust the tilt angle or curvature of the umbrella-shaped shield according to experimental requirements to further optimize fluid distribution and ensure uniform diffusion of culture medium within the chamber.
[0057] In a preferred embodiment, the biological filtration membrane 5 is positioned in a filtration module 7 .
[0058] The filtration module 7 has a first port 701 and a second port 702, each of which is detachably connected to the connecting pipe 3. Specifically, the filtration module 7 is a separate component that houses the biological filtration membrane 5 and is connected to both ends of the connecting pipe 3 via the first port 701 and the second port 702. The connection method can adopt a threaded interface, a snap-fit interface, or a quick connector to ensure sterility and stability during the connection process.
[0059] In actual operation, the user can conveniently remove the filtration module 7 from the connecting pipe 3 through the disassembly port to replace or clean the biological filtration membrane 5. This not only simplifies the maintenance process of the device, but also allows the user to select a biological filtration membrane 5 with a suitable pore size or material according to different experimental requirements, thereby flexibly adapting to a variety of cell co-culture scenarios, while ensuring the hygiene and reliability of the device during long-term use.
[0060] like Figure 3 As shown, in a preferred embodiment, the filtration module 7 includes a filter membrane box 703 composed of an upper shell and a lower shell that can be detachably matched with each other, at least one porous support plate 704 arranged inside the filter membrane box 703, and at least one elastic sealing ring 705 arranged in the filter membrane box 703.
[0061] Specifically, the membrane filter box 703 is composed of an upper shell and a lower shell, and the two are detachably connected by a threaded or snap-fit mechanism, which is convenient for the user to open the membrane filter box 703 to replace the biological filter membrane 5. The porous support plate 704 is located inside the membrane filter box 703, close to the biological filter membrane 5, to provide mechanical support to prevent the filter membrane from deforming or rupturing under fluid pressure. The elastic sealing ring 705 is installed on the joint surface or the edge of the filter membrane of the membrane filter box 703, and seals the biological filter membrane 5 through a compression action to ensure that the culture medium can only flow through the membrane pores to avoid lateral leakage. Through this structure, the filter module 7 can provide convenient operability while ensuring the consistency of the filtering effect. The user can easily complete the installation, replacement and cleaning of the filter membrane to meet the diverse needs in the experiment.
[0062] like Figure 2 As shown, in a preferred embodiment, a spherical buffer cavity 8 is provided in series in the connecting pipe 3, and the spherical buffer cavity 8 is located between the first culture chamber 1 and the filtration module 7. Specifically, the spherical buffer cavity 8 is a hollow spherical container, which is connected in series with the connecting pipe 3 through a pipe interface, and its internal space serves as a buffer area for the culture medium. When the culture medium flows from the first culture chamber 1 to the filtration module 7, the spherical buffer cavity 8 can effectively slow down the flow rate, smooth the fluid pulsation, and reduce the impact on the biological filtration membrane 5 caused by pressure fluctuations or high flow rates. Therefore, the integrity of the filter membrane is protected, its service life is extended, and the flow rate of the culture medium entering the filtration module 7 can be made more stable, thereby improving the filtration efficiency and ensuring that the cytokine-rich culture medium can be evenly and reliably transferred to the second culture chamber 2.
[0063] In a preferred embodiment, the upper portion of the spherical buffer cavity 8 has a gas phase space 801 for collecting gas, and a hydrophobic breathable membrane 802 is provided on the top of the gas phase space 801 to allow the collected gas to discharge from the buffer cavity.
[0064] Specifically, during operation, the inner space of the spherical buffer cavity 8 is filled with liquid culture medium at the bottom, and a certain gas phase space 801 is retained at the top for collecting gases that may appear in the system, such as carbon dioxide produced by cell metabolism or air remaining in the pipeline.
[0065] A hydrophobic, breathable membrane 802 is installed at the top of gas phase space 801. This membrane allows gas to pass through but blocks liquid from escaping, thereby enabling automatic gas discharge and preventing bubble accumulation in connecting pipe 3 or second culture chamber 2. The spherical buffer chamber 8 not only acts as a fluid buffer but also effectively maintains the system's gas-liquid balance, ensuring stable culture medium flow and continuously optimizing the cell culture environment, preventing bubbles from interfering with the growth of producer cells.
[0066] Of course, to minimize the impact of the external environment on the cell culture system, when the device is equipped with a hydrophobic breathable membrane, it is recommended that the entire device be placed in a biosafety cabinet or similar controlled environment. A biosafety cabinet can provide a sterile and dust-free working environment, effectively preventing the ingress of external contaminants, thereby protecting the purity of cell culture and the reliability of experimental results.
[0067] In a preferred embodiment, the communication line 3 has a valve or a pipe clamp configured to cut off or open the communication state of the communication line 3.
[0068] Specifically, the valve or clamp is positioned at an appropriate location on the connecting line 3, for example, between the first culture chamber 1 and the spherical buffer chamber 8, or between the filtration module 7 and the second culture chamber 2. The valve can be a manual knob valve or a sliding valve, while the clamp can be an adjustable clamping device. By operating the valve or clamp, the user can conveniently control the open and close state of the connecting line 3, thereby isolating the two culture chambers during specific experimental stages (such as cell inoculation, culture medium replacement, or sampling). This control mechanism enhances the operational flexibility of the device, avoids unnecessary material exchange or cross-contamination, and ensures that the user can precisely adjust culture conditions according to experimental requirements.
[0069] In a preferred embodiment, the second culture chamber 2 has a sampling port 9 , which is sealed by a detachable sealing cover.
[0070] Specifically, the sampling port 9 is arranged on the side wall or top of the second culture chamber 2 and is connected to the interior of the chamber through a sterile interface. The sampling port 9 is covered with a detachable sealing cover, such as a threaded cover, a snap cover or an elastic rubber stopper, to maintain the airtightness and sterility of the chamber when not sampling. During the sampling operation, the user can aseptically open the sealing cover, extract the culture medium or cell sample through the sampling port 9, and then reseal the sampling port 9. This facilitates the regular monitoring of the cell status or culture medium components in the second culture chamber 2 during the experiment without affecting the stability of the culture environment inside the chamber. It is particularly suitable for studies that require dynamic observation of macrophage growth or functional polarization.
[0071] Although the embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations may be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A multi-chamber cell culture device for macrophage co-culture, characterized in that: include: a first culture chamber configured for seeding engineered cells; a second culture chamber configured for seeding producer cells; a communication pipe, wherein both ends of the communication pipe are respectively connected to the first culture chamber and the second culture chamber to establish a fluid passage; Wherein, the port of the communication pipeline in the second culture chamber is configured as a ring distributor; The annular distributor is arranged along the inner circumferential wall of the second culture chamber and maintains a preset distance from the bottom surface of the second culture chamber; A plurality of through holes are provided on the inner side wall of the annular distributor; Furthermore, a biological filtration membrane for isolating cells is provided inside the communicating pipe.
2. The multi-chamber cell culture device for macrophage co-culture according to claim 1, characterized in that: The axial directions of the plurality of through holes formed on the inner side wall of the annular distributor are set to be horizontally directed toward the center of the second culture chamber, or inclined downward at a preset angle to the horizontal plane.
3. The multi-chamber cell culture device for macrophage co-culture according to claim 2, characterized in that: include: an anti-disturbance wing, arranged below the annular distributor; Wherein, the anti-disturbance wing is configured to block the medium flowing out of the through hole.
4. The multi-chamber cell culture device for macrophage co-culture according to claim 3, characterized in that: The anti-disturbance wing portion is an umbrella-shaped shield with a downwardly inclined upper surface in a conical or arc shape.
5. The multi-chamber cell culture device for macrophage co-culture according to claim 1, characterized in that: The biological filtration membrane is positioned in a filtration module; Furthermore, the filter module has a first port and a second port; The first port and the second port are respectively detachably connected to the communication pipe.
6. The multi-chamber cell culture device for macrophage co-culture according to claim 5, characterized in that: The filtering module includes: A filter membrane box composed of an upper shell and a lower shell that can be detachably matched with each other; at least one porous support plate disposed inside the membrane cartridge and configured to support the biological filtration membrane; and at least one elastic sealing ring disposed in the filter membrane box and configured to compress and seal the edge of the biological filter membrane.
7. The multi-chamber cell culture device for macrophage co-culture according to claim 6, characterized in that: A spherical buffer cavity is arranged in series in the communication pipeline, and the spherical buffer cavity is located between the first culture chamber and the filtration module.
8. The multi-chamber cell culture device for macrophage co-culture according to claim 7, characterized in that: The upper portion of the spherical buffer cavity is provided with a gas phase space for collecting gas, and a hydrophobic breathable membrane is provided on the top of the gas phase space to allow the collected gas to be discharged from the buffer cavity.
9. The multi-chamber cell culture device for macrophage co-culture according to claim 1, characterized in that: The connecting pipeline has: The valve or pipe clamp is configured to cut off or open the connection state of the connecting pipeline.
10. The multi-chamber cell culture device for macrophage co-culture according to claim 9, characterized in that: The second culture chamber has: The sampling port is sealed by a detachable sealing cover.