Cell culture platform

Abstract

This utility model relates to the field of cell culture technology and discloses a cell culture platform. The cell culture platform includes a bottom module and a top module. The bottom module includes a frame and culture components. The frame surrounds a construction area, which has multiple first inlets and outlets. The culture components are located within the construction area and include multiple culture chambers that are interconnected to form a co-culture system. The co-culture system is connected to the outside environment through the first inlets and outlets. The top of each culture chamber has an opening, and the arrangement of these openings within the construction area corresponds to the well arrangement of at least one standard well plate. The top module includes a cover plate, which is mounted on the frame and covers the construction area. This utility model adapts the cell culture platform to high-throughput instruments and equipment, facilitating efficient and convenient culture experiments.

Description

Technical Field

[0001] This utility model relates to the field of cell culture exposure technology, specifically to a cell culture platform. Background Technology

[0002] Multi-organ models are cell culture models that integrate several different tissue cells into a closed fluid network using a co-culture device. Compared with traditional two-dimensional cell models, multi-organ models can more realistically simulate the tissue-tissue interactions experienced by drugs and their related metabolites in the human body in vitro, and provide a cell microenvironment with fluid shear forces. This bridges the gap between two-dimensional cell models and living organs, and improves the consistency of drug safety and efficacy between cell-based research and preclinical trials.

[0003] In related technologies, the culture device for multi-organ models mainly uses porous membranes to make independent units, and then uses power pumps and pipelines to connect the independent units in series to build a co-culture system, generating a microenvironment with fluid shear force, which can better simulate the existence of various cell cultures that are both relatively independent and mutually interfering.

[0004] However, the connection and placement of each independent unit is cumbersome. When conducting multiple dose-effect studies, it is necessary to construct multiple co-culture systems to carry out independent experiments, which increases the number of operation steps and experimental cycle.

[0005] Furthermore, the resulting co-culture system has the problems of large footprint and irregular shape, making it unsuitable for commonly used laboratory multichannel pipettes (such as 8-channel and 12-channel pipettes) and other high-throughput instruments based on standard multiwell plates (6-well, 12-well, 24-well, 96-well, 384-well, etc.). It requires the use of single-channel pipettes to collect samples and then place them back into the multiwell plate for detection, which not only increases the experimental steps and cycle, but also makes it difficult to directly detect indicators related to adherent cells, thus increasing the difficulty of detection and reducing the application scenarios. Utility Model Content

[0006] In view of this, the present invention provides a cell culture platform to solve the problem that existing co-culture systems are difficult to adapt to high-throughput instruments and equipment.

[0007] This invention provides a cell culture platform, including a bottom module and a top module. The bottom module includes a frame and a culture component. The frame surrounds and forms a construction area. The construction area is provided with multiple first inlets and outlets. The culture component is disposed within the construction area and includes multiple culture chambers. The culture chambers are interconnected to form a co-culture system. The co-culture system is connected to the outside through the first inlets and outlets. The top of each culture chamber is provided with an opening. The arrangement of the openings within the construction area corresponds to the well arrangement of at least one standard well plate. The top module includes a cover plate disposed on the frame and covering the construction area.

[0008] Beneficial effects: Multiple culture chambers in the culture assembly are connected together to form a co-culture system. The co-culture system is connected to an external power pump through the first inlet and outlet to form a fluid circulation loop for co-culturing cells in the culture chambers. The cover plate is used to cover and protect the culture environment of the culture chambers, while the opening is used for the delivery and acquisition of materials in the culture chambers. The frame forms a construction area with a certain range and regular shape, and the arrangement of the openings in the construction area corresponds to the well position arrangement of the standard well plate, so that the cell culture platform is compatible with high-throughput instruments and equipment, which is conducive to conducting related culture experiments efficiently and conveniently.

[0009] In one alternative implementation, the culture component is adapted to change the connectivity of the culture chamber and / or the specifications of the culture chamber to form different co-culture systems.

[0010] Beneficial effects: By changing the connection method and specifications of the culture chamber, fluid routes with different flow field characteristics can be generated, thereby forming different co-culture systems to specifically simulate the interaction between different tissues and organs in the human body and meet different experimental needs.

[0011] In one alternative embodiment, the culture assembly includes a plurality of independent and detachable culture units, each culture unit having at least one culture chamber, and different numbers of the culture units are adapted to be connected to each other to form the culture assembly to change the communication mode of the culture chambers.

[0012] Beneficial effects: By disassembling the culture components into independent and detachable culture units, it is possible to flexibly change the connection method and specifications of the culture chamber according to design needs, build the required co-culture system, and meet different experimental requirements.

[0013] In one optional embodiment, the frame is provided with a first male connector and a first female connector, the first male connector and the first female connector being respectively provided with the first inlet and outlet; the culture unit is provided with a second male connector and a second female connector, the second male connector and the second female connector being respectively provided with the second inlet and outlet communicating with the culture chamber; wherein, the first male connector is adapted to be inserted into the first female connector and the second female connector, and the second male connector is adapted to be inserted into the first female connector and the second female connector.

[0014] Beneficial effects: When constructing a co-culture system, multiple culture units are connected together by plug-in connection, thereby realizing quick disassembly and assembly between culture units. The frame and culture units adopt a compatible male and female connector design, which also realizes quick disassembly and assembly between culture components and frame, improving the ease of disassembly and assembly, and helping users to flexibly construct the required fluid routes according to different experimental needs, making the construction of co-culture system more convenient and efficient.

[0015] In one optional implementation, the bottom module further includes a limiting grid plate disposed on the frame, the limiting grid plate separating the construction area to position each of the culture units.

[0016] Beneficial effects: By using limiting grids to delineate different blocks in the construction area, the culture units are placed according to the blocks, so that the position of the culture units can be more accurately adapted to high-throughput instruments and equipment, which facilitates the conduct of experiments.

[0017] In one alternative implementation, at least two of the culture chambers have different specifications, including at least one of the following: the communication method of the culture chambers; the number and location of the openings; and the shape and size of the culture chambers.

[0018] Beneficial effects: By introducing culture chambers of different specifications, users can design and build fluid routes in a personalized manner to meet the differentiated needs of different experiments.

[0019] In one alternative embodiment, the culture assembly includes a plurality of culture chambers connected in series from largest to smallest; or, the culture assembly includes a plurality of culture chambers connected in parallel.

[0020] Beneficial effects: Multiple culture chambers connected in series from large to small are suitable for simulating situations where the number of different cells in a co-culture system varies by orders of magnitude, while culture chambers connected in parallel are suitable for simulating situations where the metabolites of upstream cells act on multiple groups of cells simultaneously. This expands the types of co-culture systems that the cell culture platform can construct, meeting the differentiated needs of different experiments.

[0021] In one alternative implementation, the culture component and the frame are integrally formed.

[0022] Beneficial effects: When the co-culture system required for the experiment is already determined in advance, the culture components and the frame can be integrally formed by additive manufacturing and other methods, which can make the various culture chambers more reliably connected, reduce the risk of fluid leakage in the fluid path, and also eliminate the step of assembling and building the culture components, which facilitates the rapid development of the experiment.

[0023] In one alternative embodiment, the culture assembly includes a culture membrane disposed within the culture chamber, dividing the culture chamber into an upper culture chamber and a lower culture chamber.

[0024] Beneficial effects: By introducing a culture membrane, cells can be cultured simultaneously in the upper and lower culture chambers. The culture membrane is suitable for cell adhesion and growth, and its permeability allows for the exchange of substances between the two sides of the membrane, thus facilitating the construction of more diverse in vitro organ models and meeting the differentiated needs of different experiments.

[0025] In an optional embodiment, a middle layer module is further included, which is disposed between the top layer module and the bottom layer module. The middle layer module includes a nesting dish, which is disposed in the culture chamber through the opening. The nesting dish includes a cylindrical body and a culture membrane, with the culture membrane sealing one end of the cylindrical body. The cylindrical body and the culture membrane together form a culture chamber.

[0026] Beneficial effects: By placing the culture membrane on the middle layer module, which is independent of the bottom layer module, the culture membrane can be flexibly loaded and unloaded. This helps to flexibly configure the number and position of the culture membrane according to different co-culture systems, so as to meet the differentiated needs of different experiments.

[0027] In one alternative embodiment, the nesting dish further includes a first flange that protrudes from the outer surface of the cylinder, and the nesting dish is suspended in the culture chamber via the first flange.

[0028] Beneficial effects: By adopting a suspended setting, the culture membrane at the bottom of the nested dish can be kept suspended in the culture chamber, which allows the fluid to flow smoothly over the lower surface of the culture membrane and avoids the culture membrane from contacting the bottom of the culture chamber, thus reducing the contact with the fluid.

[0029] In one alternative embodiment, the edge of the opening supports the first flange, or the inner wall of the culture chamber is provided with a second flange that supports the first flange.

[0030] Beneficial effects: Nested dishes can be suspended at the opening or on the second flange inside the culture chamber, allowing users to flexibly adjust the setting of nested dishes according to design needs and build co-culture systems more flexibly.

[0031] In one alternative embodiment, the first flange has an injection hole that communicates with the culture chamber.

[0032] Beneficial effects: By setting up the injection port, it is convenient to directly add test substances to the culture chamber or remove liquid. When adding test substances through the fluid route, the liquid flows through each culture chamber in sequence. At this time, the cells closer to the fluid route inlet will come into contact with the test substances earlier. Direct addition can make the cells preferentially contact the more concentrated test substances.

[0033] In one alternative embodiment, the nesting dish further includes a support post disposed at the same end of the cylindrical body as the culture membrane, the support post contacting the bottom of the culture chamber to support the cylindrical body.

[0034] Beneficial effects: Nested dishes can also be placed directly in the culture chamber using support pillars, allowing fluid to flow between the pillars without affecting the interaction between the fluid and the cells on the culture membrane.

[0035] In one optional embodiment, the cover plate has an exposure hole, the position of which corresponds to the position of the opening, and the exposure hole is used to expose gas to the culture chamber or the culture room.

[0036] Beneficial effects: Specific gases can be introduced into the culture chamber or culture room through the exposure hole, creating an environment exposed to gases and meeting the needs of gas-liquid interface exposure experiments.

[0037] In one alternative embodiment, the top-layer module further includes a seal disposed on the side of the cover plate facing the assembly area, the seal sealing the gap between the top-layer module and the middle-layer module, or the seal sealing the gap between the top-layer module and the bottom-layer module.

[0038] Beneficial effects: The seal prevents gas leakage, allowing for more precise gas exposure to the target.

[0039] In one alternative embodiment, the number of seals is multiple, and the seals surround the exposure hole.

[0040] Beneficial effects: Setting up multiple seals to seal different exposure holes helps to achieve selective or differentiated exposure of gases, meeting further experimental needs. Attached Figure Description

[0041] To more clearly illustrate the specific embodiments of this utility model or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0042] Figure 1 This is a top view schematic diagram of a cell culture platform according to an embodiment of the present invention, which shows a construction mode of a co-culture system;

[0043] Figure 2 This is a top view schematic diagram of a cell culture platform according to an embodiment of the present invention, which shows another construction mode of a co-culture system;

[0044] Figure 3 This is a top view schematic diagram of a cell culture platform according to an embodiment of the present invention, which shows another construction mode of a co-culture system;

[0045] Figure 4 This is a top view schematic diagram of a cell culture platform according to an embodiment of the present invention, which shows another construction mode of a co-culture system;

[0046] Figure 5 This is a schematic diagram of the structure of the culture unit according to an embodiment of the present invention, showing the connection method of the culture unit and the arrangement of the nested dishes in the culture unit;

[0047] Figure 6 This is a schematic diagram of the structure of a culture unit according to an embodiment of the present invention, which shows another specification of the culture unit;

[0048] Figure 7 This is a schematic diagram of the structure of a culture unit according to an embodiment of the present invention, showing another specification of the culture unit;

[0049] Figure 8 This is a schematic diagram of the structure of a nested dish according to an embodiment of the present utility model;

[0050] Figure 9 This is a schematic diagram of another nested dish according to an embodiment of the present utility model;

[0051] Figure 10 This is a schematic diagram of the structure of another nested dish according to an embodiment of the present utility model;

[0052] Figure 11 This is a schematic diagram of the structure of a cell culture platform with a cover plate according to an embodiment of the present invention;

[0053] Figure 12 This is a schematic diagram of the exposed cover plate structure according to an embodiment of the present utility model;

[0054] Figure 13 This is a schematic diagram of the structure of a cell culture platform according to an embodiment of the present invention, showing the installation method of the limiting grid plate on the frame.

[0055] Explanation of reference numerals in the attached figures:

[0056] 101. Frame; 1011. Upper frame wall; 1012. Lower frame wall; 102. First male connector; 103. First female connector; 104. Third flange; 2. Culture unit; 201. Culture chamber; 202. Second male connector; 203. Second female connector; 204. Second flange; 3. Nesting dish; 301. Culture chamber; 302. Cylinder; 303. Culture membrane; 304. First flange; 305. Filling hole; 306. Support; 4. Cover plate; 401. Exposure hole; 402. Outer cover; 403. Sealing element; 5. Limiting grid plate. Detailed Implementation

[0057] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, not all embodiments. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of this utility model.

[0058] It should be understood that the terminology used herein is for the purpose of describing particular exemplary embodiments only and is not intended to be limiting. Unless the context clearly indicates otherwise, the singular forms "a," "an," and "comprising" as used herein may also mean including the plural forms. The terms "comprising," "including," and "having" are inclusive and therefore indicate the presence of the stated features, elements, and / or components, but do not exclude the presence or addition of one or more other features, elements, components, and / or combinations thereof.

[0059] Although terms such as "first," "second," etc., may be used in this document to describe multiple elements, components, regions, layers, and / or segments, these elements, components, regions, layers, and / or segments should not be limited by these terms. These terms may be used only to distinguish one element, component, region, layer, or segment from another. Unless the context clearly indicates otherwise, terms such as "first," "second," and other numerical terms used herein do not imply order or sequence. Furthermore, in the description of this application, unless otherwise expressly specified and limited, the terms "set up" and "connected" should be interpreted broadly; for example, they may refer to a fixed connection, a detachable connection, or an integral connection; they may refer to a direct connection or an indirect connection via an intermediate medium. Those skilled in the art will understand the specific meaning of the above terms in this application based on the specific circumstances.

[0060] For ease of description, spatial relative terms may be used in the text to describe the relationship of one element or feature relative to another element or feature, as shown in the figure. These relative terms include, for example, "end," "length," "inner," "outer," etc. Such spatial relative terms are intended to include different orientations of the mechanism in use or operation, in addition to those depicted in the figure. For example, if the mechanism in the figure is flipped, an element described as "below other elements or features" or "below other elements or features" would subsequently be oriented as "above other elements or features" or "above other elements or features." Therefore, the example term "below" can include both upper and lower orientations. The mechanism may be otherwise oriented (rotated 90 degrees or in other directions), and the spatial relative descriptors used in the text will be interpreted accordingly.

[0061] In related technologies, multi-organ models mainly connect independent culture chambers in series using power pumps to form a co-culture system. The power pumps enable fluid to flow between the culture chambers, which can better simulate the independent yet crosstalking existence of various cell cultures.

[0062] However, existing multi-organ models are generally personalized products, and the assembly process involves connecting and placing various culture chambers, which is cumbersome. When conducting multiple dose-effect studies, multiple sets of multi-organ models need to be assembled to carry out independent experiments, which increases the number of operation steps and experimental cycles.

[0063] Furthermore, as a customized product, it lacks standardized connection methods, and the locations of various culture chambers are not standardized. In terms of adding and removing liquids and conducting related tests on various co-culture systems, it cannot be adapted to the high-throughput instruments and equipment commonly used in laboratories that are compatible with standard multiwell plates, resulting in inconvenience in use.

[0064] The following is combined with Figures 1 to 13 The following describes embodiments of the present invention.

[0065] Reference Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 11 According to an embodiment of the present invention, a cell culture platform is provided, including a bottom module and a top module. The bottom module includes a frame 101 and a culture component. The frame 101 surrounds and forms a construction area. The construction area is provided with multiple first inlets and outlets. The culture component is disposed in the construction area and includes multiple culture chambers 201. The culture chambers 201 are interconnected to form a co-culture system. The culture chambers 201 are connected to the outside through the first inlets and outlets. The top of the culture chambers 201 is provided with an opening. The arrangement of the openings in the construction area corresponds to the arrangement of the well positions of the standard well plate. The top module includes a cover plate 4, which is disposed on the frame 101 and covers the construction area.

[0066] Multiple culture chambers 201 in the culture assembly are connected together to form a co-culture system. The co-culture system is connected to an external power pump via a first inlet and outlet to form a fluid circulation loop. In use, cells are cultured in the culture chamber 201. The cover plate 4 covers and protects the culture environment of the culture chamber 201. Under the action of the power pump, the fluid flows along the fluid path, thereby simulating the independent yet crosstalking existence of multiple cell cultures.

[0067] According to the cell culture platform of this utility model, the opening is used for material delivery and acquisition in the culture chamber 201. The frame 101 forms a construction area with a certain range and regular shape. The arrangement of the openings in the construction area corresponds to the arrangement of the well positions of the standard well plate, so that the cell culture platform is compatible with high-throughput instruments and equipment, which is conducive to efficient and convenient culture experiments.

[0068] Because high-throughput instruments can be used to collect samples directly from the cell culture platform for testing, the loss of test information caused by sample or cell transfer can be avoided, and continuous full-cycle data can be obtained.

[0069] Specifically, in order to be compatible with high-throughput instruments and equipment, the frame 101 can adopt the same size as standard multi-well plates (6-well, 12-well, 24-well, 96-well, 384-well, etc.).

[0070] For example, in Figure 1 In the embodiment, the size of the frame 101 is consistent with that of the 24-well standard multiwell plate. When constructing a co-culture system consisting of four culture chambers 201, the four culture chambers 201 can be arranged in a row, and each culture chamber 201 is provided with an opening. The position of each opening is accurately aligned with the position of the hole in the 24-well standard multiwell plate.

[0071] In some embodiments, the culture component is adapted to change the connection method and / or specifications of the culture chamber 201 to form different co-culture systems. By changing the connection method and specifications of the culture chamber 201, fluid routes with different flow field characteristics can be generated, thereby forming different co-culture systems to specifically simulate the interactions between different tissues and organs in the human body and meet different experimental needs.

[0072] For example, Figure 1 , Figure 2 , Figure 3 , Figure 4 Each of these methods demonstrates a feasible way to construct a co-culture system. The following section combines... Figure 1 , Figure 2 , Figure 3 , Figure 4 Please provide an explanation.

[0073] Reference Figure 1 , Figure 2 , Figure 3 Optionally, the culture assembly includes multiple independent and detachable culture units 2, each culture unit 2 being provided with at least one culture chamber 201, and different numbers of culture units 2 being adapted to be connected to each other to form a culture assembly, thereby changing the connection mode of the culture chambers 201.

[0074] The culture components are divided into independent and detachable culture units 2, which helps to flexibly change the connection method and specifications of the culture chamber 201 according to design needs, build the required co-culture system, and meet different experimental requirements.

[0075] Different culture units 2 can be detachably connected through various methods such as pipe connection and plug connection. As long as the stable arrangement of culture units 2 can be ensured, the arrangement of the openings and the holes of the standard well plate can correspond.

[0076] In some embodiments, the culture units 2 are detachably connected by plug-in connections. Exemplarily, the frame 101 is provided with a first male connector 102 and a first female connector 103, each with a first inlet / outlet; the culture unit 2 is provided with a second male connector 202 and a second female connector 203, each with a second inlet / outlet; wherein the first male connector 102 is adapted to be plugged into the first female connector 103 and the second female connector 203, and the second male connector 202 is adapted to be plugged into the first female connector 103 and the second female connector 203.

[0077] The first inlet and outlet are located at the first male connector 102 and the first female connector 103. Therefore, when the first male connector 102 and the first female connector 103 are plugged in, the two culture chambers 201 are also interconnected through the first inlet and outlet. When constructing the co-culture system, multiple culture units 2 are connected together by plugging in, thereby realizing quick disassembly and assembly between culture units 2. The frame 101 and the culture unit 2 adopt a compatible male and female connector design, which also realizes quick disassembly and assembly between the culture components and the frame 101, improving the ease of disassembly and assembly, and helping users to flexibly construct the required fluid routes according to different experimental needs (for example, when the preferred experimental parameters are not yet determined, the fluid route can be extended or shortened, or the direction of the fluid route can be adjusted), making the construction of the co-culture system more convenient and efficient.

[0078] Furthermore, the mutual insertion of male and female connectors also helps to achieve mutual positioning between culture units 2, making the arrangement of culture units 2 more standardized and neat.

[0079] Reference Figure 4 Alternatively, in some other embodiments, the culture component and the frame 101 are integrally formed.

[0080] The integrated co-culture system construction method is suitable for situations where the co-culture system required for the experiment has been determined in advance. By using additive manufacturing (such as photopolymerization printing) to integrate the culture components and the frame 101, the various culture chambers 201 can be more reliably connected, reducing the risk of fluid leakage in the fluid path. At the same time, it can also eliminate the step of assembling and building the culture components, which is convenient for the rapid development of the experiment.

[0081] Back Figure 1 , Figure 2 , Figure 3 It can be noted that in order to construct diverse co-culture systems, different connection methods can be used between culture chambers 201 to form different fluid routes.

[0082] In some embodiments, at least two culture chambers 201 have different specifications, including at least one of the following: the connection method of the culture chambers 201 (which may be reflected in the number and location of the second inlet and outlet); the number and location of the openings; and the shape and size of the culture chambers 201. By introducing culture chambers 201 with different specifications, users can design and construct fluid routes in a personalized manner to meet the differentiated needs of different experiments.

[0083] Optionally, in some embodiments, the culture assembly includes a plurality of culture chambers 201 connected in series from largest to smallest; or, the culture assembly includes a plurality of groups of culture chambers 201 connected in parallel.

[0084] Multiple culture chambers 201 connected in series from largest to smallest are suitable for simulating situations where the number of different cells in a co-culture system varies by orders of magnitude. Culture chambers 201 connected in parallel are suitable for simulating situations where the metabolites of upstream cells act on multiple groups of cells simultaneously, thereby expanding the types of co-culture systems that the cell culture platform can construct and meeting the differentiated needs of different experiments.

[0085] For example, refer to Figure 2 The co-culture system consists of six culture units 2. One of the culture units 2 has two second male connectors 202 and one second female connector 203. The other has one second male connector 202 and two second female connectors 203, thus forming a fluid route with parallel branches (that is, forming two sets of parallel connected culture chambers 201), which is suitable for simulating the parallel transport of tissues in the body through blood, such as the transport of substances metabolized by liver cells to downstream organs through blood in parallel.

[0086] For example, refer to Figure 3 The co-cultivation system consists of three cultivation units 2, which can be connected by pipe joints and hoses. The three cultivation units 2 are of different sizes and are connected in descending order of size, thereby further expanding the application scenarios.

[0087] For example, in studies on the effects of inhaling smoke through the mouth and tongue (sensing smoke) to the lungs (lung injury research) and then acting on the liver (hepatotoxicity and liver metabolic activation), since the number of taste bud cells in the tongue is relatively small, while the number of lung and liver cells is relatively large, the relatively larger culture unit 2 in culture chamber 201 can be selected to culture the taste bud cells, while the relatively smaller culture unit 2 in culture chamber 201 can be selected to culture the lung and liver cells. This simulates the cell composition of various organs in the body, making the experimental tests more accurate and compatible with high-throughput instruments and equipment.

[0088] Furthermore, in Figure 3 In the cell culture platform shown, three co-culture systems are constructed side-by-side within the construction area, which facilitates the replication of experiments. Culture chambers 201 of the same size are arranged in a row, and the arrangement of the openings in each row corresponds to the well arrangement of a standard plate (e.g., one well for a standard 96-well, 24-well, and 6-well plate, respectively), thus making it suitable for pairing with corresponding high-throughput instruments and equipment.

[0089] It should be pointed out that, in addition to Figure 1 , Figure 2 , Figure 3 , Figure 4 Beyond the fluid pathways shown, the cell culture platform can construct even more diverse fluid pathways using culture units of different sizes 2, which will not be elaborated upon here. Furthermore, in Figure 4In the illustrated embodiment, by replacing different underlying modules, the culture chamber 201 of the cell culture platform can also adopt different connection methods.

[0090] In some embodiments, the bottom of the culture unit 2 is made of a transparent material to facilitate observation by microscopes and other instruments.

[0091] In some embodiments, the surfaces of the second male connector 202 and / or the second female connector 203 may be provided with a sealing structure (e.g., a sealing gasket made of rubber coating) to enhance the sealing effect of the connection and prevent liquid leakage.

[0092] In some embodiments, the cell culture platform further includes a middle layer module disposed between the top layer module and the bottom layer module. The middle layer module includes a nested dish 3, which is disposed within the culture chamber 201 through an opening. By placing the nested dish 3 within the culture chamber 201, the space of the culture chamber 201 is divided into two parts (within the nested dish 3 and outside the nested dish 3). Cells can be cultured within the nested dish 3, thereby enabling the construction of more diverse in vitro organ models to meet the differentiated needs of different experiments.

[0093] Specifically, the nested dish 3 has a cylindrical body 302 and a culture membrane 303. The culture membrane 303 closes one end of the cylindrical body 302, and the cylindrical body 302 and the culture membrane 303 together form a culture chamber 301. The culture membrane 303 is suitable for cell adhesion and growth. The culture membrane 303 is permeable and has a certain pore size and pore density, thus allowing material exchange to occur inside the nested dish 3 (culture chamber 301) and outside the nested dish 3 (culture chamber 201), which is suitable for studying the effects of crosstalk between two non-contact cell types.

[0094] Furthermore, by placing the culture membrane 303 on the middle layer module, which is independent of the bottom layer module, the culture membrane 303 can be flexibly loaded and unloaded. This helps to flexibly configure the number and position of the culture membrane 303 according to different co-culture systems, so as to meet the differentiated needs of different experiments.

[0095] For example, lung cells can be cultured in culture chamber 301, and hepatocytes in culture chamber 201. When culture chamber 301 is exposed to flue gas, the flue gas can act on the lung cells and cross the simulated lung cell barrier (i.e., culture membrane 303) to enter culture chamber 201, further acting on the hepatocytes. Then, after the hepatocytes metabolize and activate the components in the flue gas, the resulting products can continue to act on the lung cells through culture membrane 303. This can be used to study the effects of in vivo inhaled exposure (flue gas, drugs) on the liver, and the effects on the lungs after activation by liver metabolism.

[0096] The nested dish 3 can be arranged in the culture chamber 201 in different ways. In some embodiments, the nested dish 3 further includes a first flange 304, which protrudes from the outer surface of the cylinder 302. The nested dish 3 is suspended in the culture chamber 201 by the first flange 304. By adopting a suspended arrangement, the culture membrane 303 at the bottom of the nested dish 3 can be ensured to be suspended in the culture chamber 201. This allows the fluid to flow smoothly over the lower surface of the culture membrane 303 and also prevents the culture membrane 303 from contacting the bottom of the culture chamber 201 and being damaged.

[0097] exist Figure 5 and Figure 8 In the illustrated embodiment, the first flange 304 is located at the top of the cylinder 302 and contacts the upper surface of the culture unit 2, thereby achieving suspension. In other embodiments, the first flange 304 may also be located at other positions on the cylinder 302, for example, see reference to Figure 7 and Figure 9 The first flange 304 can also be located in the middle of the cylinder 302. In this case, the inner sidewall of the culture chamber 201 can be provided with a second flange 204, which supports the first flange 304 to achieve suspension.

[0098] It is understandable that, in addition to the suspension arrangement, in some other embodiments, a support column 306 can also be provided at the bottom of the nested dish 3 to place the nested dish 3 in the culture chamber 201 through bottom support. Specifically, refer to... Figure 10 The nested dish 3 includes a cylindrical body 302, a culture membrane 303, and support columns 306. The cylindrical body 302 extends axially, and the culture membrane 303 closes one end of the cylindrical body 302 to form a culture chamber 301. The support columns 306 and the culture membrane 303 are located at the same end of the cylindrical body 302, and the support columns 306 are in contact with the bottom of the culture chamber 301 to support the cylindrical body 302. Fluid can flow through the spaces between the support columns 306 without affecting the interaction between the fluid and the cells on the culture membrane 303.

[0099] Additionally, refer to Figure 5 , Figure 6 , Figure 7 In a cell culture platform, a culture unit 2 may have a culture chamber 201, and a culture chamber 201 may have an opening, so that nested dishes 3 correspond one-to-one with culture unit 2. Alternatively, a culture unit 2 may have multiple culture chambers 201 and / or a culture chamber 201 may have multiple openings, so that multiple nested dishes 3 are placed in the same culture unit 2.

[0100] For example in Figure 6 In the illustrated embodiment, two nested dishes 3 are placed within the same culture chamber 201. Figure 7In the illustrated embodiment, the culture unit 2 may also have two culture chambers 201 connected together, and each chamber holds a corresponding nested dish 3. The culture unit 2 can be designed according to the needs of the actual experiment, which will not be described in detail here.

[0101] Furthermore, in some embodiments, multiple nested dishes 3 can be connected together to form a nested plate with multiple culture chambers 301. The nested plate helps to ensure the relative positional relationship between the nested dishes 3 and avoids incorrect placement of the nested dishes 3. At the same time, the nested plate also facilitates the batch placement of nested dishes 3 to the bottom module, simplifying the operation steps.

[0102] Reference Figure 5 and Figure 8 In some embodiments, the first flange 304 has a filling hole 305 that connects to the culture chamber 201. By providing the filling hole 305, it is convenient to directly add the test substance to the culture chamber 201 or remove the liquid.

[0103] For example, in related technologies, the test substance is added via a fluid route. Since the liquid flows sequentially through each culture chamber 201, cells closer to the fluid route inlet come into contact with the test substance earlier. Direct addition allows cells in a designated culture unit 2 to preferentially contact a more concentrated test substance. Another example is that, in related technologies, the nesting dish 3 typically needs to be removed before adding / removing culture medium (test substance) or cell suspension. However, with the addition well 305, culture medium (test substance) or cell suspension can be added directly at a specific point without removing the nesting dish 3, ensuring that the culture medium (test substance) is mainly present at the bottom of that specific culture chamber 201, or that cells are seeded at the bottom of that specific culture chamber 201.

[0104] The filling well 305 can exist in different ways, as long as it exposes the culture chamber 201 below the nested dish 3. Specifically... Figure 8 In the illustrated embodiment, the first flange 304 extends to form a flange-shaped annulus, and a hole is formed in the first flange 304 to form a filling hole 305, while... Figure 5 In another nested dish 3 shown, the first flange 304 extends to form a cantilever, and multiple cantilevers are arranged circumferentially on the cylinder 302, with the gap between adjacent cantilever forming a filling hole 305.

[0105] In some embodiments, the cell culture platform may omit the middle layer module and instead include a culture membrane within the culture assembly. The culture membrane is positioned within the culture chamber 201, dividing the culture chamber 201 into an upper culture chamber and a lower culture chamber. This also allows for the construction of more diverse in vitro organ models to meet the differentiated needs of various experiments.

[0106] In some embodiments, the frame 101 can be designed as a stepped structure. The frame 101 includes an upper frame wall 1011 and a lower frame wall 1012. The upper frame wall 1011 is smaller than the lower frame wall 1012 and is used to support and place the cover plate 4. The lower frame wall 1012 is used to set the first male connector 102 and the first female connector 103. A limiting step is formed between the upper frame wall 1011 and the lower frame wall 1012 to prevent the cover plate 4 from accidentally blocking the first male connector 102 and the first female connector 103.

[0107] Furthermore, in some embodiments, reference is made to Figure 12 The cover plate 4 has an exposure hole 401. Through the exposure hole 401, a specific gas can be introduced into the culture chamber 301 or the culture chamber 201 to create an environment exposed to the gas, which meets the needs of gas-liquid interface exposure experiments (e.g., exposing the culture chamber 301 to flue gas).

[0108] In addition, the cover plate 4 with the exposure hole 401 makes this invention suitable for in-situ exposure of cells, thereby meeting the application scenarios of simultaneous exposure and culture, and simultaneous exposure and detection.

[0109] Specifically, when examining the biological effects of transient perception, culture, exposure, and detection need to occur simultaneously. For example, when simulating the transient perception of sweetness by taste buds and sweet-sensing cells in vitro, one can observe the transient calcium levels within the sweet-sensing cells. 2+ Changes in the signal are used to characterize whether the sweet-sensing cells have perceived sweetness.

[0110] The cover plate 4 with exposure holes 401 allows for precise gas-liquid interface exposure of the culture chamber 301, while simultaneously allowing observation of cell changes at the bottom of the cell culture platform. When studying the tongue's perception of sweet substances in cigarette smoke during smoking, the cover plate 4 exposes the smoke into the culture chamber 301, where cells can produce Ca2+ upon exposure. 2+ Fluorescence signals can be observed in real time from the bottom using microscopes, which can characterize the cells' perception of the sweetness of smoke.

[0111] For example, with Figure 12 In the illustrated embodiment, the exposure port 401 is adapted to introduce a specific gas into the culture chamber 301. The cover plate 4 also includes an outer cover 402 surrounding the outer periphery of the exposure port 401. One end of the outer cover 402 is used to connect to an external gas supply device, and the diameter of the other end of the outer cover 402 gradually increases, with the maximum diameter being smaller than the diameter of the culture chamber 301, thereby allowing the outer cover 402 to extend into the culture chamber 301 and ensuring that the introduced gas can be delivered to the location of the cells.

[0112] In some embodiments, the cover plate 4 further includes a seal 403 disposed on the side of the cover plate 4 facing the assembly area. The seal 403 seals the gap between the top layer module and the middle layer module, or the seal 403 seals the gap between the top layer module and the bottom layer module. The seal 403 can prevent gas from leaking into other areas of the co-culture system, so as to allow for more precise exposure of the target gas.

[0113] Specifically Figure 12 In the illustrated embodiment, there are multiple seals 403 surrounding the exposure holes 401. The seals 403 can contact the surface of the nested dish 3 to achieve a sealing effect. Providing multiple seals 403 to seal different exposure holes 401 facilitates selective or differentiated gas exposure, meeting further experimental requirements.

[0114] It is understandable that the seal 403 can be integrally molded on the cover plate 4, or the seal 403 can also be a sealing ring, etc., with the cover plate 4 having a sealing groove and the sealing ring embedded in the sealing groove.

[0115] In some embodiments, an exhaust port (not shown in the figure) is also provided between the seal 403 and the outer cover 402. Gas passes through the top module through the exposure hole 401 and enters the culture chamber 301 under the constraint of the outer cover 402. It blows onto the liquid in the culture chamber 301 to exchange substances between the gas and the cells. After that, the gas bypasses the outer cover 402 and leaves the culture chamber 301 through the gap between the outer cover 402 and the cylinder 302. It is then discharged through the exhaust port under the constraint of the seal 403, so that the gas can be continuously delivered.

[0116] Optionally, the bottom of the outer cover 402 is between 1 mm and 2 mm from the liquid surface, thereby improving the effect of gas exposure.

[0117] Understandably, in Figure 12 In this embodiment, the exposure hole 401 is used to expose gas to the culture chamber 301. In other embodiments, the exposure hole 401 can also expose gas to the culture chamber 201 according to a similar principle, which will not be described in detail here.

[0118] In some embodiments, the cover plate 4 is provided with a marking structure to facilitate the determination of the placement orientation of the cover plate 4. Specifically, the marking structure is located on the outer side of the cover plate 4 (that is, the surface facing away from the culture assembly). The marking structure can be various shapes suitable for identification and differentiation (such as annular protrusions, etc.) so that the user can divide the area of ​​the cover plate 4 into sections and conveniently locate the position of the culture chamber 201.

[0119] In some embodiments, the inner surface of the cover plate 4 is provided with protrusions (e.g., on...). Figure 12 (A protrusion is provided at the corner of the inner side of the cover plate 4) to allow a certain ventilation gap between the cover plate 4 and the frame 101.

[0120] Reference Figure 13 In some embodiments, the cell culture platform further includes a limiting grid 5, which is disposed on the frame 101 and divides the construction area to position each culture unit 2.

[0121] For example, the limiting grid 5 may further include transverse grids and longitudinal grids, which are arranged alternately on the frame 101. By using the limiting grid 5 to delineate different blocks in the construction area, the culture units 2 are placed according to the blocks, so that the position of the culture units 2 can be more accurately adapted to high-throughput instruments and equipment, facilitating the conduct of experiments.

[0122] Continue to refer to Figure 9 For example, the frame 101 may further include a third flange 104, which extends inward from the inner side of the frame 101. The third flange 104 is provided with a slot that mates with the limiting grid plate 5 (e.g., with an interference fit). The limiting grid plate 5 is inserted into the slot, thereby accurately placing the limiting grid plate 5. The connection parts of the transverse grid plate and the longitudinal grid plate are provided with a similar engaging structure to facilitate mutual engagement and stable placement.

[0123] It should be noted that in some embodiments, the bottom of the frame 101 is open (that is, the bottom module does not have a base plate), and the frame 101 is placed on the worktable, which facilitates the observation of the cell culture platform from below by microscopes and other optical instruments. Of course, in other embodiments, the bottom module may also include a base plate, with the frame 101 set on the base plate, and observation holes provided on the base plate.

[0124] According to an embodiment of the present invention, another aspect provides a culture method using the cell culture platform provided by the present invention, the culture method comprising:

[0125] Step S110: Construct a co-culture system in the construction area according to the preset fluid route.

[0126] For example, in some embodiments, it is necessary to select culture units 2 corresponding to 6-well, 12-well, 24-well, and 96-well plates according to the research purpose, determine the number and connection method of culture units 2, and select a frame 101 of appropriate size that is compatible with the flow channel openings (first male connector 102 and first female connector 103).

[0127] For example, when performing organoid culture, culture unit 2 with sizes corresponding to 6, 12, or 24 wells can be selected; when performing cell line culture, culture unit 2 with a size corresponding to 96 wells can be selected; and when performing organoid and cell line co-culture or cell line and reservoir circulation, culture unit 2 of various sizes can be used simultaneously.

[0128] For example, in some embodiments, the cover plate 4 may contain or partially contain the exposure hole 401 depending on whether gas-liquid interface exposure is required.

[0129] Step S120: Seed cells in the co-culture system.

[0130] Specifically, based on cell adhesion, cell culture can be categorized into adherent cell culture, suspension cell culture, and co-culture of adherent and suspension cells. Adherent cell culture includes cell adhesion culture in culture chamber 201, cell adhesion culture in culture chamber 301, and co-culture of adherent cells in culture chambers 201 and 301; suspension cell culture includes cell suspension culture in culture chamber 201, cell suspension culture in culture chamber 301, and co-culture of suspension cells in culture chambers 201 and 301.

[0131] Step S130: Supply fluid to the co-culture system via an external power pump to culture the cells.

[0132] The culture method uses the cell culture platform provided by this invention, and therefore has the beneficial effects brought by the cell culture platform, which will not be elaborated here.

[0133] Optionally, in some embodiments, cells can also be observed in situ during the culture process.

[0134] The following examples will help to better understand the cell culture platform and culture method of this invention.

[0135] Example 1: Adherent cell culture in culture chamber 201.

[0136] A suitable cell culture platform was constructed according to the research objectives. The connected cell culture platform was placed in a clean bench. Resuspended cells were seeded into culture chamber 201, which had been treated with cell adhesion promoters. Liquid was then added to culture chamber 201. Cover plate 4 was placed on frame 101, and the cell culture platform was placed in an incubator for 24 hours. Cells and their adhesion were then observed. Based on the cell culture results, subsequent procedures included continued cell culture, observation, medium changes, passage, or exposure experiments.

[0137] Example 2: Co-culture of adherent cells in culture chamber 201 and culture chamber 301.

[0138] The difference between this example and Example 1 is that after adding liquid to culture chamber 201, nested dish 3 is placed inside culture chamber 201, and the resuspended cells are seeded into culture chamber 301, which has been treated with cell adhesion promoters, and then cultured. At this point, the liquid level should be higher than the culture membrane 303 of nested dish 3, and the pore size of culture membrane 303 should generally not allow cells to pass through the pores, such as being set to ≤6μm. Other aspects not mentioned in this example can be found in Example 1 and will not be repeated here.

[0139] Co-adherent culture can be used to study the effects of crosstalk between two non-contact cells, such as the effects of in vivo inhaled exposure (smoke, drugs) on the liver, and the effects on the lungs after activation by liver metabolism.

[0140] Example 3: Suspension cell culture in culture chamber 201.

[0141] The difference between this example and Example 1 is that it is not necessary to add cell adhesion promoters before seeding cells. Other details not mentioned in this example can be found in Example 1 and will not be repeated here.

[0142] Suspension culture refers to the growth of cells suspended in a culture medium, which is suitable for culturing immune cells and macrophages in the blood. These cells travel with the blood and body fluids in vivo, while in the in vitro culture environment they are suspended in a culture medium and move with the flow of the culture medium or migrate on their own according to stimuli.

[0143] Example 4: Co-culture of suspension cells in culture chamber 201 and culture chamber 301.

[0144] The difference between this example and Example 2 is that it is not necessary to add cell adhesion promoters before seeding cells. Other details not mentioned in this example can be found in Example 2 and will not be repeated here.

[0145] In this example, when cells are allowed to penetrate culture membrane 303, the pore size of culture membrane 303 is >6μm, which is suitable for culturing macrophages. Macrophages are seeded in culture chamber 301. When a stimulating substance that promotes macrophage migration appears in culture chamber 201, the macrophages will move into culture chamber 201. Thus, the presence of these cells in different locations can be used to characterize inflammation or the generation of certain substances or related mechanisms and responses.

[0146] Example 5: Co-culture of adherent cells in culture chamber 201 and suspension cells in culture chamber 301.

[0147] The difference between this example and Example 4 is that a cell adhesion promoter needs to be added before seeding cells in culture chamber 201. Other details not mentioned in this example can be found in Example 4 and will not be repeated here.

[0148] This example is suitable for examining environments where suspended and adherent cells coexist, such as vascular cells and immune cells, or organ cells and immune cells. For instance, it can be used to conduct experiments on the recruitment of macrophages (suspended) by an inflammatory response in lung / vascular cells (adherent), where macrophages migrate to the lung / vascular cells based on the substances generated by the inflammatory response.

[0149] Example 6: Co-culture of suspension cells in culture chamber 201 and adherent cells in culture chamber 301.

[0150] The difference between this example and Example 4 is that a cell adhesion promoter needs to be added before seeding cells in culture chamber 301. Other details not mentioned in this example can be found in Example 4 and will not be repeated here.

[0151] Example 7: Suspended cells and adherent cells are co-cultured in culture chamber 201 and culture chamber 301.

[0152] The difference between this example and Example 2 is that, after 24 hours of culture, cell adhesion is observed. Suspension cells are sequentially seeded into culture chamber 201 and culture chamber 301, and cell adhesion and suspension are observed. Other details not mentioned in this example can be found in Example 2 and will not be repeated here.

[0153] Example 8: Cell culture and exposure at the gas-liquid interface.

[0154] When culturing cells at the gas-liquid interface, suspension cells are usually not cultured in culture chamber 301. The preliminary procedure can be referred to in Examples 2 and 6. The difference is that after 24 hours of inoculation and culture, the liquid in culture chamber 301 is removed, and then the cover plate 4 is placed on top. The device is then placed in an incubator for gas-liquid interface culture. Depending on the cell culture situation, subsequent observation, medium replacement, passage, or exposure experiments are carried out.

[0155] When conducting the exposure test, the cover plate 4 is replaced with a cover plate 4 having an exposure hole 401, and then the exposure hole 401 is connected to the pipe of the external exposure device for exposure.

[0156] Example 9: Co-culture of lung cells, vascular cells, hepatocytes, and macrophages.

[0157] This example uses three culture units 2 connected in series. Suspended macrophages are cultured in the culture chamber 201 of each culture unit 2. Nested dishes 3 are set in each culture unit 2, and adherent lung cells, vascular cells and hepatocytes are cultured in the culture chamber 301 respectively.

[0158] Example 10: Co-culture of lung cells, vascular cells, hepatocytes and macrophages in a reservoir.

[0159] The difference between this example and Example 9 is that this example adds an additional culture unit 2 to the tandem system. These culture units 2 do not require the placement of nesting dishes 3 or cell inoculation, and function as reservoirs.

[0160] Although embodiments of the present invention have been described in conjunction with the accompanying drawings, those skilled in the art can make various modifications and variations without departing from the spirit and scope of the present invention, and such modifications and variations all fall within the scope defined by the appended claims.

Claims

1. A cell culture platform, characterized in that, include: The bottom module includes a frame (101) and a culture component. The frame (101) surrounds and forms a construction area. The construction area is provided with multiple first inlets and outlets. The culture component is located in the construction area. The culture component includes multiple culture chambers (201). The culture chambers (201) are interconnected to form a co-culture system. The co-culture system is connected to the outside world through the first inlets and outlets. The top of the culture chamber (201) is provided with an opening, and the arrangement of the opening in the construction area corresponds to the arrangement of the well positions of at least one standard well plate. The top-level module includes a cover plate (4), which is disposed on the frame (101) and covers the construction area.

2. The cell culture platform according to claim 1, characterized in that, The culture component is adapted to change the connection mode of the culture chamber (201) and / or the specifications of the culture chamber (201) to form different co-culture systems.

3. The cell culture platform according to claim 2, characterized in that, The culture assembly includes multiple independent and detachable culture units (2), each culture unit (2) having at least one culture chamber (201). Different numbers of the culture units (2) are adapted to be connected to each other to form the culture assembly, thereby changing the connection mode of the culture chamber (201).

4. The cell culture platform according to claim 3, characterized in that, The frame (101) is provided with a first male connector (102) and a first female connector (103), and the first male connector (102) and the first female connector (103) are respectively provided with the first inlet and outlet; The culture unit (2) is provided with a second male connector (202) and a second female connector (203), and the second male connector (202) and the second female connector (203) are respectively provided with a second inlet and outlet communicating with the culture chamber (201); The first male connector (102) is adapted to be inserted into the first female connector (103) and the second female connector (203), and the second male connector (202) is adapted to be inserted into the first female connector (103) and the second female connector (203).

5. The cell culture platform according to claim 3, characterized in that, The bottom module also includes a limiting grid plate (5), which is disposed on the frame (101) and separates the construction area to position each of the cultivation units (2).

6. The cell culture platform according to claim 2, characterized in that, At least two of the culture chambers (201) have different specifications, and the specifications include at least one of the following: The connection method of the culture chamber (201); The number and location of the openings; The shape and dimensions of the culture chamber (201).

7. The cell culture platform according to claim 6, characterized in that, The culture assembly includes a plurality of culture chambers (201) connected in series from largest to smallest; Alternatively, the culture assembly may include a plurality of culture chambers (201) connected in parallel.

8. The cell culture platform according to claim 1, characterized in that, The culture component and the frame (101) are integrally formed.

9. The cell culture platform according to any one of claims 1 to 8, characterized in that, The culture assembly includes a culture membrane disposed within the culture chamber (201) and dividing the culture chamber (201) into an upper culture chamber and a lower culture chamber.

10. The cell culture platform according to any one of claims 1 to 8, characterized in that, It also includes a middle layer module, which is disposed between the top layer module and the bottom layer module. The middle layer module includes a nested dish (3), which is disposed within the culture chamber (201) through the opening. The nested dish (3) includes a cylindrical body (302) and a culture membrane (303). The cylindrical body (302) is axially continuous, and the culture membrane (303) closes one end of the cylindrical body (302). The cylindrical body (302) and the culture membrane (303) together form a culture chamber (301).

11. The cell culture platform according to claim 10, characterized in that, The nesting dish (3) further includes a first flange (304), which protrudes from the outer surface of the cylinder (302), and the nesting dish (3) is suspended in the culture chamber (201) through the first flange (304).

12. The cell culture platform according to claim 11, characterized in that, The edge of the opening supports the first flange (304), or the inner wall of the culture chamber (201) is provided with a second flange (204), which supports the first flange (304).

13. The cell culture platform according to claim 11, characterized in that, The first flange (304) has an injection hole (305) that is connected to the culture chamber (201).

14. The cell culture platform according to claim 10, characterized in that, The nested dish (3) also includes a support (306), The support column (306) and the culture membrane (303) are located at the same end of the cylinder (302), and the support column (306) is in contact with the bottom of the culture chamber (201) to support the cylinder (302).

15. The cell culture platform according to claim 10, characterized in that, The cover plate (4) has an exposure hole (401) which is located in a position corresponding to the position of the opening. The exposure hole (401) is used to expose gas to the culture chamber (301) or the culture chamber (201).

16. The cell culture platform according to claim 15, characterized in that, The top-level module also includes a seal (403), which is disposed on the side of the cover plate (4) facing the assembly area. The seal (403) seals the gap between the top-level module and the middle-level module, or the seal (403) seals the gap between the top-level module and the bottom-level module.

17. The cell culture platform according to claim 16, characterized in that, The number of seals is multiple, and the seals (403) surround the exposure hole (401).

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