Multi-cell co-culture chip structure
By designing the multi-cell co-culture chip structure, the limitations of Transwell chambers in cell co-culture are solved, and interactions and information exchange between multiple cells are achieved, providing an effective platform for the study of drug action mechanisms.
Patent Information
- Application Number
- CN202510085908.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-20
- Publication Date
- 2025-04-25
AI Technical Summary
The existing Transwell chambers have limitations in cell coculture, which cannot communicate between multiple cells or tissues, and are not conducive to direct microscopic observation and imaging detection.
A multi-cell co-culture chip structure is designed, including a transparent cell culture plate and multiple cell culture chambers, with microchannels set up between any two cell culture chambers, allowing free exchange of liquids and adding specific stimulation factors to each cell culture chamber.
The interaction and information exchange between any two cells are achieved, providing a platform for studying the mechanism of action of drugs between multiple organs, and supporting dynamic co-culture of multiple cells, establishing a more bionic in vitro model.
Smart Images
Figure CN119875830A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of microfluidic chips, and more specifically, to a multi-cell co-culture chip structure. Background Art
[0002] The interaction between cells is a complex process and is of great significance for maintaining homeostasis. The interaction between various types of cells in the human body is achieved through direct cell-cell contact, cell-extracellular matrix interaction, secretion of bioactive factors and extracellular vesicles, thereby affecting the morphology, vitality, proliferation, differentiation and function of cells. The cell co-culture system is used as an in vitro research model in biomedical research, which can better restore the microenvironment in the body and provide a powerful tool for understanding cell interactions and tumor metastasis and invasion.
[0003] The types of cell co-culture can be divided into indirect co-culture and direct co-culture. Direct co-culture is when multiple cells are inoculated in the same well at the same time or separately, and different types of cells are in direct contact with each other. Indirect co-culture is when multiple cells are inoculated on different carriers and then placed in the same culture environment, and different types of cells are not in direct contact with each other. Indirect co-culture can be achieved by using microcarriers, scaffolds, double-layer membranes, and specific culture media, and is used to study cell-dependent interactions without physical contact. At present, Transwell chambers are considered to be able to simulate the pathological and physiological processes of human tissues and organs to a large extent, and are widely used in conventional cell co-culture experiments. In the Transwell co-culture system, one type of cell is cultured at the bottom of the Transwell chamber, while another type of cell is cultured on the upper porous membrane. The porous membrane of the Transwell acts as a tissue barrier to separate the two cells, and allows small molecules secreted by the upper and lower cells to pass through, realizing indirect co-culture of the two cells. The advantage of using Transwell for cell co-culture is that the technology is simple, but the Transwell chamber can only be cultured statically, and communication between multiple cells or tissues cannot be carried out. It is also not conducive to direct microscopic observation and imaging detection, and has certain limitations. Summary of the invention
[0004] In order to overcome the above-mentioned shortcomings, the present invention provides a multi-cell co-culture chip structure, which realizes the interaction between any two cells in the co-culture model and allows the addition of specific stimulation factors in each cell culture chamber, thereby realizing information exchange between cells in each channel and providing a platform for studying the mechanism of action of drugs between multiple organs.
[0005] In order to solve the above technical problems, the present invention adopts the following technical solution: a multi-cell co-culture chip structure, including a transparent cell culture plate, on which a plurality of cell culture chambers are arranged, and a microchannel is arranged between any two cell culture chambers to allow free exchange of liquids in all cell culture chambers.
[0006] Before cell culture, disinfect with 75% alcohol and sterilize with ultraviolet irradiation. Then slowly inject multiple cells into multiple cell culture chambers at appropriate cell densities. When the cell suspension fills the cell culture chamber, stop injecting the cell suspension. After observing the cell suspension in the chamber under a microscope to ensure that it is evenly distributed and leak-free, place the chip in a cell culture incubator at 37°C and 5% CO2 for 2-4 hours. After the cells adhere to the wall, use a precision injection pump to continuously inject the culture solution corresponding to each cell at a rate of 2.0 μL / min for dynamic co-culture of multiple cells.
[0007] The cell culture plate is equipped with multiple cell culture chambers, which can be injected with a variety of different cell suspensions for cell culture, and microchannels are set between any two cell culture chambers, so that all cell culture chambers are interconnected, enabling free flow and exchange of liquids. The transparent cell culture plate facilitates optical imaging detection of cell culture to obtain high-resolution sample images, and in-situ detection based on the live cell imaging system can provide more instantaneous signals of cell activity.
[0008] The multi-cell co-culture chip structure of this patent application realizes the interaction between any two cells in the co-culture model and allows the addition of specific stimulation factors in each cell culture chamber, thereby realizing information exchange between cells in each channel and providing a platform for studying the mechanism of action of drugs between multiple organs.
[0009] Preferably, the surface of the cell culture plate is sunken to form a cell culture chamber, and a transparent cover is connected to the cell culture plate, which covers the cell culture chamber; a liquid inlet and a liquid outlet are respectively arranged at corresponding positions on the cover and at both ends of the cell culture chamber.
[0010] The cover plate and the cell culture plate are tightly connected to achieve the closure of the cell culture chamber, inject cell suspension and culture fluid through the liquid inlet, and discharge the culture fluid through the liquid outlet to achieve multi-cell dynamic co-culture. The transparent cover plate facilitates optical imaging detection.
[0011] Preferably, a cell culture tank is arranged on the cover plate, the cell culture tank is arranged toward the cell culture plate, the opening of the cell culture tank is connected to the porous membrane, and all the cell culture chambers overlap with the porous membrane.
[0012] A cell culture tank is arranged on the cover plate, which adds a cell culture chamber. The porous membrane serves as a substrate for the cells in the upper cell culture tank to adhere to the wall and grow, and allows information exchange between the upper and lower cell culture tanks and cell culture chambers to achieve dynamic co-culture of multiple cells.
[0013] Preferably, the plurality of cell culture chambers are evenly distributed around the circumference.
[0014] Multiple cell culture chambers are evenly distributed around the circumference to facilitate the layout of microchannels.
[0015] Preferably, the microchannel comprises a plurality of microgrooves arranged at intervals, and the microgrooves communicate between the cell culture chambers.
[0016] Multiple microgrooves arranged at intervals form microchannels. The microgrooves are smaller than the diameter of the cells, ensuring the free flow and exchange of liquids between different cell culture chambers. The size of the microgrooves is micrometer-level, and the microchannels allow the free exchange of secretions such as cytokines, making it possible to co-culture multiple cells.
[0017] Preferably, the width of the cell culture chamber is 500-1000 μm, and the depth of the cell culture chamber is 80-120 μm.
[0018] The size of the cell culture chamber is in the micron level, which can not only reduce the consumption of reagents to lower costs, but also match the physiological microenvironment in the human body well.
[0019] Preferably, the cell culture chamber comprises a first chamber and a second chamber, the first chamber and the second chamber are connected together to form a V-shaped structure with an obtuse corner, the three cell culture chambers are arranged on the cell culture plate to form a "Y"-shaped structure, and microchannels are arranged between the first chamber and the second chamber of the cell culture chamber and the two adjacent cell culture chambers.
[0020] In this solution, three cell culture chambers are arranged on the cell culture plate to form a "Y"-shaped structure, and the angle between the first chamber and the second chamber of the cell culture chamber is 120 degrees, which can be used for dynamic co-culture of three types of cells. Microchannels are arranged between the first chamber and the second chamber of the cell culture chamber and the two adjacent cell culture chambers, thereby realizing the connection between any two cell culture chambers, which is conducive to the free flow and exchange of liquids between different cell culture chambers.
[0021] In another embodiment, the cell culture chamber includes a first chamber and a second chamber, the first chamber and the second chamber are connected together to form a V-shaped structure with an acute angle, and three cell culture chambers are arranged on a cell culture plate to form a "△"-shaped structure, and microchannels are arranged between the first chamber and the second chamber of the cell culture chamber and the two adjacent cell culture chambers.
[0022] In this solution, three cell culture chambers are arranged on the cell culture plate to form a "△"-shaped structure, and the angle between the first chamber and the second chamber of the cell culture chamber is 60 degrees, which can be used for dynamic co-culture of three types of cells. Microchannels are arranged between the first chamber and the second chamber of the cell culture chamber and the two adjacent cell culture chambers, thereby realizing the connection between any two cell culture chambers, which is conducive to the free flow and exchange of liquids between different cell culture chambers.
[0023] Preferably, the liquid flows in the cell culture chambers on both sides of the microchannel in the same direction.
[0024] The liquid flow direction in the cell culture chambers on both sides of the microchannel is consistent to avoid the opposite direction of liquid flow, which may cause convection of the liquid connecting the two sides of the microchannel during continuous perfusion, causing physical damage to the cells or reducing the efficiency of material exchange in the liquid.
[0025] Preferably, multiple cell culture chambers are injected with multiple cell suspensions, and after the cells adhere to the wall, the culture fluid corresponding to each cell is continuously injected to perform multi-cell dynamic co-culture.
[0026] In this way, dynamic co-culture of multiple cells is achieved, which helps to establish a more bionic in vitro model and provide a more accurate in vitro model experimental platform for physiological and pathological research, toxicology research and drug screening.
[0027] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) The multi-cell co-culture chip structure of the present patent application realizes the interaction between any two cells in the co-culture model, and allows the addition of specific stimulation factors in each cell culture chamber, thereby realizing the information exchange between cells in each channel, and providing a platform for studying the mechanism of action of drugs between multiple organs; (2) The multi-cell co-culture chip of the present patent application realizes the dynamic co-culture of multiple cells, which helps to establish a more bionic in vitro model, and provides a more accurate in vitro model experimental platform for physiological and pathological research, toxicological research and drug screening; (3) The multi-cell co-culture chip provides a separate culture channel for each cell, and is provided with a separate liquid inlet and outlet, so that each type of cell can be subjected to separate drug intervention and post-sampling biochemical detection, allowing optical imaging detection of each type of cell sample. BRIEF DESCRIPTION OF THE DRAWINGS
[0028] Figure 1 It is a schematic structural diagram of embodiment 1 of the present invention.
[0029] Figure 2 It is an exploded view of Example 1 of the present invention.
[0030] Figure 3 It is a schematic structural diagram of embodiment 2 of the present invention.
[0031] Figure 4 It is an exploded view of Example 2 of the present invention.
[0032] Figure 5 It is a schematic structural diagram of embodiment 3 of the present invention.
[0033] Figure 6 It is an exploded view of Example 3 of the present invention.
[0034] Figure 7 It is a partial structural diagram of Example 4 of the present invention.
[0035] In the figure: 1. cell culture plate, 2. cell culture chamber, 3. cover plate, 4. liquid inlet, 5. liquid outlet, 6. microchannel, 7. microgroove, 8. micro-column, 9. first chamber, 10. second chamber, 11. narrowed channel, 12. liquid level, 13. liquid mixing section, 14. cell culture tank, 15. porous membrane, 16. inlet, 17. outlet, 18. flow guide, 19. slide plate, 20. through groove, 21. magnetic strip, 22. slide groove, 23. mounting groove. DETAILED DESCRIPTION
[0036] The technical solution of the present invention is further described in detail below through specific embodiments and in conjunction with the accompanying drawings: Example 1: A multi-cell co-culture chip structure (see Figure 1 , Figure 2 ), comprising a transparent cell culture plate 1, on which a plurality of cell culture chambers 2 are arranged, the surface of the cell culture plate 1 is sunken to form the cell culture chambers 2, and the cell culture plate 1 is connected to a transparent cover plate 3, which covers the cell culture chambers 2; the cell culture plate 1 and the cover plate 3 are both made of polydimethylsiloxane, which has high transparency, heat resistance, cold resistance, small viscosity change with temperature, waterproofness, small surface tension, good electrical insulation, weather resistance, hydrophobicity, and good biocompatibility.
[0037] A plurality of cell culture chambers 2 are evenly distributed circumferentially, and a liquid inlet 4 and a liquid outlet 5 are respectively arranged at corresponding positions on the cover plate 3 and at both ends of the cell culture chamber 2. A microchannel 6 is arranged between any two cell culture chambers 2 to allow free exchange of liquids in all cell culture chambers 2. The microchannel 6 includes a plurality of microgrooves 7 arranged at intervals, and the microgrooves 7 are connected between the cell culture chambers 2. The microgrooves 7 are 10 μm, which is smaller than the diameter of the cells, thereby ensuring the free flow and exchange of liquids between different cell culture chambers 2. A row of microcolumns 8 is arranged between adjacent cell culture chambers 2, and the spacing between adjacent microcolumns 8 forms microgrooves 7, and the microcolumns 8 are columnar structures with rectangular cross-sections.
[0038] The width of the cell culture chamber 2 is 500-1000 μm, and the depth of the cell culture chamber 2 is 80-120 μm. In this embodiment, the width of the cell culture chamber 2 is 800 μm, and the depth of the cell culture chamber 2 is 100 μm. The size of the cell culture chamber 2 is micrometer-level, which can not only reduce the consumption of reagents to reduce costs, but also match the physiological microenvironment in the human body. Multiple cell culture chambers 2 are injected with multiple cell suspensions, and the culture fluid corresponding to each cell is continuously injected after the cells adhere to the wall to perform multi-cell dynamic co-culture.
[0039] The cell culture plate 1 and the cover plate 3 are both rectangular structures, and the corners of the cell culture plate 1 and the cover plate 3 are rounded. The cell culture chamber 2 includes a first chamber 9 and a second chamber 10, which are connected together to form a V-shaped structure with an obtuse corner. The three cell culture chambers 2 are arranged on the cell culture plate 1 to form a "Y"-shaped structure. The first chamber 9 and the second chamber 10 of the cell culture chamber 2 are respectively arranged close to the two adjacent cell culture chambers, and microchannels 6 are arranged between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 and the two adjacent cell culture chambers. Narrowing channels 11 are arranged at both ends of the cell culture chamber 2, and the width of the narrowing channel 11 is smaller than the width of the cell culture chamber 2. The liquid inlet 4 and the liquid outlet 5 arranged on the cover plate 3 are arranged corresponding to the ends of the narrowing channel 11, and the end of the cell culture chamber 2 and the narrowing channel 11 are gradually narrowed.
[0040] The ends of two adjacent cell culture chambers 2 are close together to form three liquid levels 12, wherein the two cell culture chambers 2 of one liquid level 12 are respectively connected with two liquid inlets 4, the two cell culture chambers 2 of another liquid level 12 are respectively connected with two liquid outlets 5, and the two cell culture chambers 2 of the third liquid level 12 are respectively connected with one liquid outlet 5 and one liquid inlet 4. The three cell culture chambers 2 are close together to form a "Y"-shaped structure, thereby forming three liquid mixing sections 13, which are connected together in a Y shape. By arranging the liquid outlets 5 and the liquid inlets 4 at both ends of the cell culture chambers 2, the two liquid flows in two liquid mixing sections 13 flow in the same direction, and the two liquid flows in the other liquid mixing section 13 flow in the opposite direction, thereby reducing the impact damage to the cells caused by the reverse flow of liquid.
[0041] The angle between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 is 120 degrees, which can be used for dynamic co-culture of three types of cells. Microchannels 6 are provided between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 and the two adjacent cell culture chambers, thereby achieving communication between any two cell culture chambers 2, which is conducive to the free flow and exchange of liquids between different cell culture chambers.
[0042] Before cell culture, disinfect with 75% alcohol and sterilize with ultraviolet irradiation. Then, multiple cells are slowly injected into multiple cell culture chambers 2 at appropriate cell densities. When the cell suspension fills the cell culture chamber 2, stop injecting the cell suspension. After observing the uniform distribution of the cell suspension in the chamber under a microscope and no leakage, place the chip in a cell culture incubator at 37°C and 5% CO2 for 2-4 h. After the cells adhere to the wall, use a precision injection pump to continuously inject the culture solution corresponding to each cell at a rate of 2.0 μL / min for dynamic co-culture of multiple cells.
[0043] Three cell culture chambers 2 are arranged on the cell culture plate 1, into which three different cell suspensions can be injected for cell culture, and a microchannel 6 is arranged between any two cell culture chambers 2, so that all the cell culture chambers 2 are interconnected, and free flow exchange of liquids can be achieved. The transparent cell culture plate 1 and cover plate 3 facilitate optical imaging detection of cell culture to obtain high-resolution sample images, and in-situ detection based on the live cell imaging system can provide more instantaneous signals of cell activities.
[0044] Example 2: A multi-cell co-culture chip structure (see Figure 3 , Figure 4 ), comprising a transparent cell culture plate 1, on which a plurality of cell culture chambers 2 are arranged, the surface of the cell culture plate 1 is sunken to form the cell culture chambers 2, and the cell culture plate 1 is connected to a transparent cover plate 3, which covers the cell culture chambers 2; the cell culture plate 1 and the cover plate 3 are both made of polydimethylsiloxane, which has high transparency, heat resistance, cold resistance, small viscosity change with temperature, waterproofness, small surface tension, good electrical insulation, weather resistance, hydrophobicity, and good biocompatibility.
[0045] A plurality of cell culture chambers 2 are evenly distributed circumferentially, and a liquid inlet 4 and a liquid outlet 5 are respectively arranged at corresponding positions on the cover plate 3 and at both ends of the cell culture chamber 2. A microchannel 6 is arranged between any two cell culture chambers 2 to allow free exchange of liquids in all cell culture chambers 2. The microchannel 6 includes a plurality of microgrooves 7 arranged at intervals, and the microgrooves 7 are connected between the cell culture chambers 2. The microgrooves 7 are 10 μm, which is smaller than the diameter of the cells, thereby ensuring the free flow and exchange of liquids between different cell culture chambers 2. A row of microcolumns 8 is arranged between adjacent cell culture chambers 2, and the spacing between adjacent microcolumns 8 forms microgrooves 7, and the microcolumns 8 are columnar structures with rectangular cross-sections.
[0046] The width of the cell culture chamber 2 is 500-1000 μm, and the depth of the cell culture chamber 2 is 80-120 μm. In this embodiment, the width of the cell culture chamber 2 is 800 μm, and the depth of the cell culture chamber 2 is 100 μm. The size of the cell culture chamber 2 is micrometer-level, which can not only reduce the consumption of reagents to reduce costs, but also match the physiological microenvironment in the human body. Multiple cell culture chambers 2 are injected with multiple cell suspensions, and the culture fluid corresponding to each cell is continuously injected after the cells adhere to the wall to perform multi-cell dynamic co-culture.
[0047] The cell culture plate 1 and the cover plate 3 are both triangular structures, and the corners of the cell culture plate 1 and the cover plate 3 are rounded. The cell culture chamber 2 includes a first chamber 9 and a second chamber 10. The first chamber 9 and the second chamber 10 are connected together to form a V-shaped structure with an acute corner. The connection between the first chamber 9 and the second chamber 10 is rounded. The angle between the first chamber 9 and the second chamber 10 is 60 degrees. The three cell culture chambers 2 are arranged on the cell culture plate 1 to form a "△" shaped structure. The first chamber 9 and the second chamber 10 of the cell culture chamber 2 are respectively arranged close to the two adjacent cell culture chambers, and the first chamber 9 and the second chamber 10 of the cell culture chamber 2 are arranged between the two adjacent cell culture chambers. Microchannels 6 are arranged between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 and the two adjacent cell culture chambers. The liquid flow direction in the cell culture chambers 2 on both sides of the microchannel 6 is consistent.
[0048] Both ends of the cell culture chamber 2 are provided with narrowed channels 11, the width of the narrowed channel 11 is smaller than the width of the cell culture chamber 2, the liquid inlet 4 and the liquid outlet 5 provided on the cover plate 3 are provided corresponding to the ends of the narrowed channel 11, and the ends of the cell culture chamber 2 and the narrowed channel 11 are gradually narrowed. The three cell culture chambers 2 are close together to form a "△"-shaped structure, thereby forming a three-stage mixed liquid section 13, and the three-stage mixed liquid sections 13 are connected together to form a "△" shape. By arranging the liquid outlet 5 and the liquid inlet 4 at both ends of the cell culture chamber 2, the two liquid flows in the three-stage mixed liquid section 13 flow in the same direction, avoiding the impact damage to the cells caused by the reverse flow of liquid.
[0049] In this solution, three cell culture chambers 2 are arranged on the cell culture plate 1 to form a "△" shaped structure, and the angle between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 is 60 degrees, which can be used for dynamic co-culture of three types of cells. Microchannels 6 are arranged between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 and the two adjacent cell culture chambers, thereby realizing the connection between any two cell culture chambers 2, which is conducive to the free flow and exchange of liquids between different cell culture chambers.
[0050] Before cell culture, disinfect with 75% alcohol and sterilize with ultraviolet irradiation. Then, multiple cells are slowly injected into multiple cell culture chambers 2 at appropriate cell densities. When the cell suspension fills the cell culture chamber 2, stop injecting the cell suspension. After observing the uniform distribution of the cell suspension in the chamber under a microscope and no leakage, place the chip in a cell culture incubator at 37°C and 5% CO2 for 2-4 h. After the cells adhere to the wall, use a precision injection pump to continuously inject the culture solution corresponding to each cell at a rate of 2.0 μL / min for dynamic co-culture of multiple cells.
[0051] Three cell culture chambers 2 are arranged on the cell culture plate 1, into which three different cell suspensions can be injected for cell culture, and a microchannel 6 is arranged between any two cell culture chambers 2, so that all the cell culture chambers 2 are interconnected, and free flow exchange of liquids can be achieved. The transparent cell culture plate 1 and cover plate 3 facilitate optical imaging detection of cell culture to obtain high-resolution sample images, and in-situ detection based on the live cell imaging system can provide more instantaneous signals of cell activities.
[0052] Example 3: A multi-cell co-culture chip structure (see Figure 5 , Figure 6 ), comprising a transparent cell culture plate 1, on which a plurality of cell culture chambers 2 are arranged, the surface of the cell culture plate 1 is sunken to form the cell culture chambers 2, and the cell culture plate 1 is connected to a transparent cover plate 3, which covers the cell culture chambers 2; the cell culture plate 1 and the cover plate 3 are both made of polydimethylsiloxane, which has high transparency, heat resistance, cold resistance, small viscosity change with temperature, waterproofness, small surface tension, good electrical insulation, weather resistance, hydrophobicity, and high shear resistance.
[0053] A plurality of cell culture chambers 2 are evenly distributed circumferentially, and a liquid inlet 4 and a liquid outlet 5 are respectively arranged at corresponding positions on the cover plate 3 and at both ends of the cell culture chamber 2. A microchannel 6 is arranged between any two cell culture chambers 2 to allow free exchange of liquids in all cell culture chambers 2. The microchannel 6 includes a plurality of microgrooves 7 arranged at intervals, and the microgrooves 7 are connected between the cell culture chambers 2. The microgrooves 7 are 10 μm, which is smaller than the diameter of the cells, thereby ensuring the free flow and exchange of liquids between different cell culture chambers 2. A row of microcolumns 8 is arranged between adjacent cell culture chambers 2, and the spacing between adjacent microcolumns 8 forms microgrooves 7, and the microcolumns 8 are columnar structures with rectangular cross-sections.
[0054] The width of the cell culture chamber 2 is 500-1000 μm, and the depth of the cell culture chamber 2 is 80-120 μm. In this embodiment, the width of the cell culture chamber 2 is 800 μm, and the depth of the cell culture chamber 2 is 100 μm. The size of the cell culture chamber 2 is micrometer-level, which can not only reduce the consumption of reagents to reduce costs, but also match the physiological microenvironment in the human body. Multiple cell culture chambers 2 are injected with multiple cell suspensions, and the culture fluid corresponding to each cell is continuously injected after the cells adhere to the wall to perform multi-cell dynamic co-culture.
[0055] The cell culture plate 1 and the cover plate 3 are both rectangular structures, and the corners of the cell culture plate 1 and the cover plate 3 are rounded. The cell culture chamber 2 includes a first chamber 9 and a second chamber 10, which are connected together to form a V-shaped structure with an obtuse corner. The three cell culture chambers 2 are arranged on the cell culture plate 1 to form a "Y"-shaped structure. The first chamber 9 and the second chamber 10 of the cell culture chamber 2 are respectively arranged close to the two adjacent cell culture chambers, and microchannels 6 are arranged between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 and the two adjacent cell culture chambers. Narrowing channels 11 are arranged at both ends of the cell culture chamber 2, and the width of the narrowing channel 11 is smaller than the width of the cell culture chamber 2. The liquid inlet 4 and the liquid outlet 5 arranged on the cover plate 3 are arranged corresponding to the ends of the narrowing channel 11, and the end of the cell culture chamber 2 and the narrowing channel 11 are gradually narrowed.
[0056] The ends of two adjacent cell culture chambers 2 are close together to form three liquid levels 12, wherein the two cell culture chambers 2 of one liquid level 12 are respectively connected with two liquid inlets 4, the two cell culture chambers 2 of another liquid level 12 are respectively connected with two liquid outlets 5, and the two cell culture chambers 2 of the third liquid level 12 are respectively connected with one liquid outlet 5 and one liquid inlet 4. The three cell culture chambers 2 are close together to form a "Y"-shaped structure, thereby forming three liquid mixing sections 13, which are connected together in a Y shape. By arranging the liquid outlets 5 and the liquid inlets 4 at both ends of the cell culture chambers 2, the two liquid flows in two liquid mixing sections 13 flow in the same direction, and the two liquid flows in the other liquid mixing section 13 flow in the opposite direction, thereby reducing the impact damage to the cells caused by the reverse flow of liquid.
[0057] The angle between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 is 120 degrees, which can be used for dynamic co-culture of three types of cells. Microchannels 6 are provided between the first chamber 9 and the second chamber 10 of the cell culture chamber 2 and the two adjacent cell culture chambers, thereby achieving communication between any two cell culture chambers 2, which is conducive to the free flow and exchange of liquids between different cell culture chambers.
[0058] A cell culture tank 14 is arranged on the cover plate 3, and the cell culture tank 14 is arranged toward the cell culture plate 1. A porous membrane 15 is connected to the opening of the cell culture tank 14, and all the cell culture chambers 2 overlap with the porous membrane 15. The material of the porous membrane 15 is a polycarbonate film or a polyester film with good light transmittance. The cell culture tank 14 is arranged on the lower surface of the cover plate 3, and an inlet 16 and an outlet 17 connected to the cell culture tank 14 are arranged on the cover plate 3. A guide channel 18 is arranged between the inlet 16, the outlet 17 and the cell culture tank 14, and the guide channel 18 and the cell culture chamber 2 are staggered. The outer contour of the porous membrane 15 is larger than the opening contour of the cell culture tank 14, and the porous membrane 15 is placed between the cell culture plate 1 and the cover plate 3.
[0059] A cell culture tank 14 is arranged on the cover plate 3, which adds a cell culture chamber. The porous membrane 15 serves as a substrate for the cells in the upper cell culture tank 14 to adhere to the wall and grow, and allows information exchange between the upper and lower cell culture tanks 14 and the cell culture chamber 2 to achieve dynamic co-culture of multiple cells.
[0060] Before cell culture, disinfect with 75% alcohol and sterilize with ultraviolet irradiation. Then, multiple cells are slowly injected into multiple cell culture chambers 2 at appropriate cell densities. When the cell suspension fills the cell culture chamber 2, stop injecting the cell suspension. After observing the uniform distribution of the cell suspension in the chamber under a microscope and no leakage, place the chip in a cell culture incubator at 37°C and 5% CO2 for 2-4 h. After the cells adhere to the wall, use a precision injection pump to continuously inject the culture solution corresponding to each cell at a rate of 2.0 μL / min for dynamic co-culture of multiple cells.
[0061] Three cell culture chambers 2 and then a cell culture tank 14 are arranged on the cell culture plate 1, and four different cell suspensions can be injected for cell culture, and a microchannel 6 is arranged between any two cell culture chambers 2, and all cell culture chambers 2 overlap with the porous membrane 15, so that all cell culture chambers 2 and cell culture tanks 14 are interconnected, and free flow exchange of liquids can be achieved. Four separate cell culture areas realize co-culture of four types of cells, thereby improving the cell sample throughput of the chip and meeting more experimental needs. The transparent cell culture plate 1 and cover plate 3 facilitate optical imaging detection of cell culture to obtain high-resolution sample images, and in situ detection based on the living cell imaging system can provide more instantaneous signals of cell activities.
[0062] Example 4: A multi-cell co-culture chip structure (see Figure 7), its structure is similar to any one of the embodiments 1 to 3, the main difference is that in this embodiment, a slide plate 19 is slidably installed on the upper cover, the slide plate 19 is correspondingly arranged with the microchannel 6, a through groove 20 is correspondingly arranged on the slide plate 19 and the microgroove 7, a magnetic strip 21 is arranged on the slide plate 19, a slide groove 22 is correspondingly arranged on the cell culture plate 1 and the microchannel 6, and the lower part of the slide plate 19 is movably installed in the slide groove 22. When the slide plate 19 slides to make the through groove 20 and the microgroove 7 communicate with each other, the microchannel 6 is connected to the adjacent two cell culture chambers 2. When the slide plate 19 slides to make the through groove 20 and the microgroove 7 misaligned, the microchannel 6 is closed and the adjacent two cell culture chambers 2 are isolated. A magnetic strip 21 is arranged on the slide plate 19, and the magnetic block is close to the magnetic strip 21 on the outer side of the cover plate 3. The movement of the magnetic block can drive the slide plate 19 to move. The slide groove 22 passes through all the micro-columns 8, and a mounting groove 23 is correspondingly arranged on the cover plate 3 and the slide plate 19. The upper part of the mounting groove 23 is slidably installed in the mounting groove 23, and the length of the mounting groove 23 is greater than the length of the slide plate 19. In this embodiment, the microchannel 6 can be opened and closed, thereby achieving the connection and isolation of two adjacent cell culture chambers 2, meeting a variety of different usage scenarios. The other structures are the same as any one of embodiments 1 to 3.
[0063] The above-described embodiments are only preferred solutions of the present invention and are not intended to limit the present invention in any form. Other variations and modifications are possible without exceeding the technical solutions described in the claims.
Claims
1. A multi-cell co-culture chip structure, characterized in that: The invention comprises a transparent cell culture plate, on which a plurality of cell culture chambers are arranged, and a microchannel is arranged between any two cell culture chambers to allow free exchange of liquids in all the cell culture chambers.
2. A multi-cell co-culture chip structure according to claim 1, characterized in that: The surface of the cell culture plate is sunken to form a cell culture chamber, and a transparent cover is connected to the cell culture plate to cover the cell culture chamber; a liquid inlet and a liquid outlet are respectively arranged at corresponding positions on the cover and at both ends of the cell culture chamber.
3. A multi-cell co-culture chip structure according to claim 2, characterized in that: A cell culture tank is arranged on the cover plate, the cell culture tank is arranged toward the cell culture plate, the opening of the cell culture tank is connected to the porous membrane, and all the cell culture chambers overlap with the porous membrane.
4. The multi-cell co-culture chip structure according to claim 1, characterized in that: A plurality of cell culture chambers are evenly distributed around the circumference.
5. The multi-cell co-culture chip structure according to claim 1, characterized in that: The microchannel includes a plurality of microgrooves arranged at intervals, and the microgrooves communicate between the cell culture chambers.
6. The multi-cell co-culture chip structure according to claim 1, characterized in that: The width of the cell culture chamber is 500-1000 μm, and the depth of the cell culture chamber is 80-120 μm.
7. The multi-cell co-culture chip structure according to claim 1, characterized in that: The cell culture chamber includes a first chamber and a second chamber, the first chamber and the second chamber are connected together to form a V-shaped structure with an obtuse corner, and the three cell culture chambers are arranged on the cell culture plate to form a "Y"-shaped structure. Microchannels are arranged between the first chamber and the second chamber of the cell culture chamber and the two adjacent cell culture chambers.
8. The multi-cell co-culture chip structure according to claim 1, characterized in that: The cell culture chamber includes a first chamber and a second chamber, the first chamber and the second chamber are connected together to form a V-shaped structure with an acute angle, and three cell culture chambers are arranged on a cell culture plate to form a "△"-shaped structure, and microchannels are arranged between the first chamber and the second chamber of the cell culture chamber and the two adjacent cell culture chambers.
9. The multi-cell co-culture chip structure according to claim 8, characterized in that: The liquid flow directions in the cell culture chambers on both sides of the microchannel are consistent.
10. A multi-cell co-culture chip structure according to any one of claims 1 to 9, characterized in that: Multiple cell culture chambers are injected with a variety of cell suspensions, and after the cells adhere to the wall, the culture fluid corresponding to each cell is continuously injected to perform multi-cell dynamic co-culture.