A self-gravity high-throughput membrane chip

Abstract

The application discloses a self-gravity high-throughput membrane chip. The membrane chip comprises a first layer, a porous membrane and a second layer. The first layer has a first culture chamber; the second layer is stacked along the gravity direction on the first layer, and at least one of the first layer and the second layer has a second culture chamber, which is located on the side of the porous membrane away from the first culture chamber in the gravity direction; the second layer has at least one first sample inlet and at least one second sample inlet, and in the gravity direction, the first sample inlet of the first sample pool and the second sample inlet of the second sample pool are both higher than the second culture chamber. The application can make the fluid enter the first culture chamber and the second culture chamber correspondingly and adhere to the corresponding surface of the porous membrane in a self-gravity fluid driving mode, simulate cell dynamic culture, improve the culture throughput, provide convenience for constructing a complex organ model, and reduce the complexity of a perfusion device and the cost of organ model construction.

Description

Technical Field

[0001] This application relates to the field of cell culture technology, and in particular to a gravity-driven high-throughput membrane chip. Background Technology

[0002] Membrane-on-a-chip (MEA) technology is a technique for three-dimensional in vitro cell culture on a chip. By constructing microchannels, microreaction chambers, and other functional components, it precisely manipulates components such as cells, fluids, gases, and the extracellular microenvironment within the chip, thereby generating biofunctional human microtissues and microorganisms. MEA technology offers advantages such as integration, low consumption, high throughput, high simulation fidelity, and rapid analysis, and has broad application prospects in fields such as new drug development, disease models, personalized medicine, and aerospace medicine.

[0003] The human body transports nutrients and gases to various parts of the body via blood vessels, and exchanges nutrients, gases, and metabolites through the organ-vascular interface. To simulate this structure, some membrane-on-a-chip (MEA) research uses porous membranes to construct such interfaces, culturing vascular endothelial cells and corresponding organ cells on either side of the membrane to simulate the cell arrangement and interface function of the organ-vascular interface. However, due to the complexity of the human body, the complexity of the models that MEA can build is highly demanding. Furthermore, the fluid within the MEA must be able to flow to better simulate the growth environment of human micro-tissues and micro-organs. This results in complex MEA structures, high costs for constructing organ models, complex construction processes, and low throughput. Summary of the Invention

[0004] This application provides a gravity-driven high-throughput membrane chip, which can solve the problem of complex membrane chip structure.

[0005] This application provides a gravity-driven high-throughput membrane chip, which includes a first layer, a porous membrane, and a second layer.

[0006] The first layer has a first space, which includes a first culture chamber; a porous membrane is disposed corresponding to the first culture chamber; a second layer is stacked on the first layer along the direction of gravity, and at least one of the first layer and the second layer has a second space, which includes a second culture chamber, and in the direction of gravity, the second culture chamber is located on the side of the porous membrane away from the first culture chamber; the second layer has at least one first injection cell and at least one second injection cell, one end of the first injection cell is connected to the first culture chamber and the other end forms a first injection port on the surface of the second layer away from the first layer, one end of the second injection cell is connected to the second culture chamber and the other end forms a second injection port on the surface of the second layer away from the first layer, and in the direction of gravity, both the first injection port and the second injection port are higher than the second culture chamber.

[0007] In some exemplary embodiments, the first space further includes a first flow channel communicating with the first inlet and the first culture chamber, the first flow channel and the first culture chamber being located in a first plane perpendicular to the direction of gravity; the second space further includes a second flow channel communicating with the second inlet and the second culture chamber, the second flow channel and the second culture chamber being located in a second plane perpendicular to the direction of gravity; both the first plane and the second plane are parallel to the porous membrane and are located on opposite sides of the porous membrane in the direction of gravity.

[0008] In some exemplary embodiments, the first space is a rotationally symmetric space or an axisymmetric space; and / or, the second space is a rotationally symmetric space or an axisymmetric space.

[0009] In some exemplary embodiments, the first layer has a first mating surface connected to the second layer, the first layer has the second space, and the second culture chamber and the second flow channel are formed on the first mating surface.

[0010] In some exemplary embodiments, the first layer includes a culture layer and a base plate stacked in the direction of gravity; the culture layer includes a first mating surface connected to the second layer and a second mating surface opposite to the first mating surface; the first culture chamber and the first flow channel are formed on the second mating surface, and the second mating surface is connected to the base plate.

[0011] In some exemplary embodiments, the edge region of the porous membrane is fixed to at least one of the first layer and the second layer; or, the membrane chip further includes a support ring, the edge region of the porous membrane is fixed to the support ring, and the support ring is mounted on at least one of the first layer and the second layer.

[0012] In some exemplary embodiments, the second layer has a first hole on its surface opposite to the first layer, and the first hole extends toward the first layer to communicate with the second culture chamber.

[0013] In some exemplary embodiments, the second layer has a plurality of evaporation tanks on its surface away from the first layer; the second layer has a plurality of first inlets and a plurality of second inlets; and at least one of the evaporation tanks is provided between two adjacent first inlets, between two adjacent second inlets, and between adjacent first inlets and second inlets.

[0014] In some exemplary embodiments, the shape of the first culture chamber is similar to that of the second culture chamber, and is a circular hole, a regular polygonal hole, or a regular polygonal rounded corner hole.

[0015] In some exemplary embodiments, the first space, the second space, the first inlet, the second inlet, and the porous membrane form a group of culture units, and the membrane chip has multiple groups of culture units, with any two groups of culture units spaced apart.

[0016] Secondly, this application also provides an application of the membrane chip described above in biological model culture and drug analysis.

[0017] The gravity-driven high-throughput film chip based on the embodiments of this application has at least the following beneficial effects:

[0018] By using gravity-driven fluid propulsion, fluid can enter and exit the first and second culture chambers respectively, adhering to the corresponding surfaces of the porous membrane. This also facilitates reciprocating motion under the drive of the propulsion device, allowing the fluid within the first and second culture chambers to flow and contact the cells attached to the porous membrane surface, simulating dynamic cell culture. Furthermore, the membrane chip eliminates the need for a separate structure to drive the fluid flow, reducing the space occupied by structures delivering fluid to the first and second culture chambers. This allows for the placement of more structural units for cell culture within a limited space, increasing culture throughput. The membrane chip of this application provides convenience for constructing complex organ models and also facilitates the establishment of standard three-dimensional organ model culture methods, reducing the complexity of perfusion equipment and the cost of organ model construction. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a three-dimensional structural diagram of a film-type chip according to an embodiment of this application;

[0021] Figure 2 This is a schematic diagram of the exploded structure of a film-type chip according to an embodiment of this application;

[0022] Figure 3 This is a three-dimensional structural diagram of a membrane chip having multiple culture units according to an embodiment of this application;

[0023] Figure 4 This is a three-dimensional structural diagram of a film-type chip with a first hole according to an embodiment of this application;

[0024] Figure 5 This is a cross-sectional schematic diagram of a film chip having a first hole according to an embodiment of this application;

[0025] Figure 6 This is a three-dimensional structural diagram of a membrane chip having a first pore and multiple culture units according to an embodiment of this application;

[0026] Figure 7 This is a three-dimensional structural diagram of a film-type chip with an evaporation tank according to an embodiment of this application;

[0027] Figure 8 This is a cross-sectional schematic diagram of a film-type chip with an evaporation tank according to an embodiment of this application;

[0028] Figure 9 This is an exploded schematic diagram of a film-type chip with an evaporation tank according to an embodiment of this application;

[0029] Figure 10 This is a three-dimensional structural diagram of a membrane chip having an evaporation tank and multiple culture units according to an embodiment of this application.

[0030] Figure label:

[0031] 10. Membrane-based chip; 11. Culture unit;

[0032] 100. First layer; 110. Culture layer; 111. First docking surface; 113. Step surface; 120. Base plate;

[0033] 101. First Space; 1011. First Incubation Chamber; 1012. First Flow Channel; 1013. Transfer Flow Channel;

[0034] 200. Second layer; 201. Second space; 2011. Second culture chamber; 2012. Second flow channel; 202. First injection port; 203. Second injection port; 204. First hole; 205. Evaporation tank; 2021. First injection cell; 2031. Second injection cell;

[0035] 300. Porous membrane;

[0036] 400. Support ring. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0038] The inventors discovered that membrane-based chips containing only one flow channel limit the types of organ models that can be cultured and are not conducive to the establishment of complex models. Membrane-based chips with both upper and lower flow channels can meet the needs of establishing complex models. However, in related technologies, membrane-based chips with upper and lower flow channels use peristaltic pumps or air pumps for fluid drive, resulting in complex and expensive devices, high costs for building organ models, complex operation during the construction process, and low throughput. In addition, regardless of the culture method (e.g., chip culture or device culture), if the requirements for high-throughput model construction cannot be met, it will be difficult to meet the needs of large-scale drug screening, severely limiting the application of this culture method. Based on this, embodiments of this application provide a gravity-driven high-throughput membrane-based chip.

[0039] like Figure 1 and Figure 2 The diagram shown is a structural schematic of a gravity-driven high-throughput membrane chip 10 according to an embodiment of this application. The membrane chip 10 includes a first layer 100, a porous membrane 300, and a second layer 200.

[0040] The first layer 100 has a first space 101, which includes a first culture chamber 1011. A second layer 200 is stacked on one side of the first layer 100, and at least one of the first layer 100 and the second layer 200 has a second space 201, which includes a second culture chamber 2011 corresponding to the first culture chamber 1011. A porous membrane 300 is mounted on at least one of the first layer 100 and the second layer 200 and is disposed corresponding to the first culture chamber 1011. The second culture chamber 2011 is located on the side of the porous membrane 300 facing away from the first culture chamber 1011, such that one surface of the porous membrane 300 can contact the material in the first culture chamber 1011, and the other surface can contact the material in the second culture chamber 2011. The porous membrane 300 covers the junction between the first culture chamber 1011 and the second culture chamber 2011 to prevent the substances in the first culture chamber 1011 and the second culture chamber 2011 from being directly exchanged without passing through the porous membrane 300.

[0041] The second layer 200, facing away from the first layer 100, has at least one first inlet 202 communicating with the first space 101 and at least one second inlet 203 communicating with the second space 201. Fluid entering the first space 101 through the first inlet 202 can reach the first culture chamber 1011 and contact the porous membrane 300, and fluid entering the second space 201 through the second inlet 203 can reach the second culture chamber 2011 and contact the porous membrane 300. For example, a cell suspension containing cells can be perfused into one of the first inlet 202 and the second inlet 203. The cell suspension enters the corresponding culture chamber, and the cells are adsorbed onto one side of the porous membrane 300 to carry out cell culture in the corresponding mode; or, cell suspensions containing different cells can be perfused into the first inlet 202 and the second inlet 203 respectively. The cell suspensions enter the corresponding culture chambers, and the cells are adsorbed onto the surface of the porous membrane 300 to carry out bilateral cell growth in the corresponding mode.

[0042] The second layer 200 has at least one first injection cell 2021, one end of which is connected to the first culture chamber 1011, and the other end of which forms a first injection port 202 on the surface of the second layer 200 opposite to the first layer 100. The second layer 200 also has at least one second injection cell 2031, one end of which is connected to the second culture chamber 2011, and the other end of which forms a second injection port 203 on the surface of the second layer 200 opposite to the first layer 100. Fluid injected through the first injection port 202 passes through the first injection cell 2021 and enters the first culture chamber 1011, while fluid injected through the second injection port 203 passes through the second injection cell 2031 and enters the second culture chamber 2011. In the direction of gravity G, the first sample injection chamber 2021 and the second sample injection chamber 2031 have depth, which on the one hand prevents fluid from splashing out, and on the other hand reserves space to store fluid, so that the first culture chamber 1011 and the second culture chamber 2011 can be filled with fluid to correspondingly wet the cells adsorbed on the surface of the porous membrane 300.

[0043] Specifically, in this embodiment, the second layer 200 is disposed on the first layer 100 along the gravity direction G. In the gravity direction G, the second culture chamber 2011 is located above the porous membrane 300, and the first culture chamber 1011 is located below the porous membrane 300. In the gravity direction G, both the first inlet 202 and the second inlet 203 are higher than the second culture chamber 2011. Thus, the fluid buffered in the first inlet chamber 2021 and the fluid buffered in the second inlet chamber 2031 can be driven by gravity to allow fluid entering the first space 101 to reach the first culture chamber 1011 and wet one surface of the porous membrane 300, and fluid entering the second space 201 to reach the second culture chamber 2011 and wet the other surface of the porous membrane 300.

[0044] When using the membrane chip 10 of this application for cell culture, the membrane chip 10 is installed in a driving device. The reciprocating motion of the membrane chip 10 is controlled by the driving device, allowing the fluids in the first culture chamber 1011 and the second culture chamber 2011 to flow and contact the cells attached to the porous membrane 300, simulating dynamic cell culture. Furthermore, by using gravity-driven fluid propulsion, the membrane chip 10 eliminates the need for a separate structure to drive the fluid flow, helping to reduce the space occupied by structures that transport fluid to the first culture chamber 1011 and the second culture chamber 2011. This allows for the placement of more structural units for cell culture within a limited space, increasing culture throughput. For example, as... Figure 3As shown, the first space 101, the second space 201, the first sample inlet 202, the second sample inlet 203 and the porous membrane 300 form a group of culture units 11. The membrane chip 10 has multiple groups of culture units 11. Any two groups of culture units 11 are arranged at intervals, and the distance between two adjacent groups of culture units 11 can be designed to be closer, so as to make full use of the space of the membrane chip 10 and effectively improve the culture throughput of the membrane chip 10.

[0045] The membrane chip 10 of this application embodiment provides convenience for constructing complex organ models, and also helps to establish a standard three-dimensional organ model culture method. Fluid can be directly perfused from the first injection port 202 and the second injection port 203, which reduces the complexity of the perfusion equipment and the cost of organ model construction.

[0046] Optionally, there may be multiple first inlet ports 202 connected to the first culture chamber 1011; and / or multiple second inlet ports 203 connected to the second culture chamber 2011. By controlling the composition of the fluid injected from each of the first inlet ports 202 and second inlet ports 203, a wide variety of organ models can be established. For example, by controlling the composition of the drug contained in the injected fluid, the needs of large-scale drug screening can be met.

[0047] To better simulate the dynamic cellular environment, the first space 101 further includes a first flow channel 1012, one end of which is directly connected to the first culture chamber 1011, and the other end is connected to the first sample inlet 2021. The second space 201 also includes a second flow channel 2012, one end of which is directly connected to the second culture chamber 2011, and the other end is connected to the second sample inlet 2031. Optionally, the flow area of ​​the first flow channel 1012 remains constant in the flow direction; or, in the flow direction towards the first culture chamber 1011, the flow area of ​​the first flow channel 1012 gradually decreases, so as to regulate the flow rate or velocity of the fluid entering the first culture chamber 1011. Optionally, the first flow channel 1012 is a strip-shaped flow channel, and the direction of extension of the first flow channel is designed to regulate the flow direction of the fluid entering the first culture chamber 1011. Similarly, the flow rate, velocity, and direction of fluid entering the second culture chamber 2011 through the second flow channel 2012 can be controlled by controlling the shape of the second flow channel 2012.

[0048] The first flow channel 1012 and the first culture chamber 1011 are located on a first plane perpendicular to the direction of gravity G, and the second flow channel 2012 and the second culture chamber 2011 are located on a second plane perpendicular to the direction of gravity G. Both the first and second planes are parallel to the porous membrane 300 and are located on opposite sides of the porous membrane 300 in the direction of gravity G. This allows the corresponding fluids to flow more smoothly and stably into and out of the first culture chamber 1011 and the second culture chamber 2011, and the flow direction of the fluids is parallel to the flattened membrane surface of the porous membrane 300. The fluids can contact the cells adsorbed on the porous membrane 300 more stably, which helps to establish a standard three-dimensional organ model culture method.

[0049] Optionally, when there are multiple first inlet ports 202, the first space 101 includes multiple first flow channels 1012, which are equal in number and correspond one-to-one with the number of first inlet ports 202. This allows the fluid in the first culture chamber 1011 to flow more smoothly through the cells adsorbed on the surface of the porous membrane 300, thereby better simulating the dynamic cell culture process. Furthermore, the multiple first flow channels 1012 directly connected to the first culture chamber 1011 are arranged in a rotationally symmetrical or axially symmetrical manner.

[0050] Optionally, when there are multiple second inlets 203, the second space 201 includes multiple second channels 2012, which are equal in number and correspond one-to-one with the number of second inlets 203, so that the fluid in the second culture chamber 2011 can flow more smoothly through the cells adsorbed on the surface of the porous membrane 300, thereby better simulating the dynamic cell culture process. Further, the multiple second channels 2012 are arranged rotationally symmetrically or axially symmetrically.

[0051] Further, the first space 101 is a rotationally symmetric space or an axisymmetric space. Specifically, the first space 101 is a rotationally symmetric space centered on the center of the first culture chamber 1011, or the first space 101 is an axisymmetric space symmetric about a plane passing through the center of the first culture chamber 1011. Optionally, the second space 201 is a rotationally symmetric space or an axisymmetric space. Specifically, the second space 201 is a rotationally symmetric space centered on the center of the second culture chamber 2011, or the second space 201 is an axisymmetric space about a plane passing through the center of the second culture chamber 2011. Thus, the first space 101 and the second space 201 have a symmetrical structure, which helps the corresponding fluid to flow more smoothly over the cells adsorbed on the surface of the porous membrane 300 when reciprocating under the drive of the driving device. For example, when the driver chip of the driving device rotates and reciprocates in a preset direction, the first space 101 and the second space 201 are each set to be rotationally symmetric spaces; or, when the driver chip of the driving device reciprocates in a straight line, the first space 101 and the second space 201 are each set to be axisymmetric spaces.

[0052] During organ-on-a-chip operation, to prevent fluid from splashing out from the first inlet 202 and the second inlet 203, it is understood that, in the direction of gravity G, the greater the distance from the first inlet 202 to the first culture chamber 1011, the better the splash prevention effect, and the greater the distance from both the second inlet 203 and the second culture chamber 2011, the better the splash prevention effect. Optionally, the second layer 200 has a second space 201. Specifically, the second space 201 is formed on the surface of the second layer 200 for connecting with the first layer 100, and a first space 101 is formed on the surface of the first layer 100 for connecting with the second layer 200. The porous membrane 300 is sandwiched between the first layer 100 and the second layer 200. Alternatively, the first layer 100 has a second space 201. Specifically, the first layer 100 has a first mating surface 111 for connecting with the second layer 200, and the second culture chamber 2011 and the second flow channel 2012 are formed on the first mating surface 111. The porous membrane 300 is disposed on the first layer 100. This arrangement places the second space 201 in a region relatively far from the first inlet 202 and the second inlet 203, providing a good anti-splashing effect.

[0053] When the first layer 100 has a second space 201, the first space 101 further includes a transition channel 1013 communicating with the first flow channel 1012. The transition channel 1013 extends to the first mating surface 111 for communicating with the first sample inlet 2021, so that when the first layer 100 and the second layer 200 are stacked, the first space 101 in the lower layer communicates with the first sample inlet 2021. For example, the transition channel 1013 extends to the first mating surface 111 along the stacking direction of the first layer 100 and the second layer 200.

[0054] like Figures 1 to 7 As shown, the first layer 100 includes a culture layer 110 and a base plate 120 stacked in the direction of gravity G; one surface of the culture layer 110 forms a first mating surface 111, and the culture layer 110 also includes a second mating surface facing away from the first mating surface 111, the second mating surface being used to connect with the base plate 120. A first culture chamber 1011 and a first flow channel 1012 are formed on the second mating surface, and the second mating surface is sealed to the base plate 120.

[0055] Optionally, when the first layer 100 has both a first space 101 and a second space 201, the second culture chamber 2011 and the second flow channel 2012 are formed on the first mating surface 111 of the culture layer 110, and the first culture chamber 1011 and the first flow channel 1012 are formed on the second mating surface of the culture layer 110. By setting the first flow channel 1012 and the second flow channel 2012 on the surface of the culture layer 110, it is convenient to process and form the culture layer 110, and at the same time, the first space 101 and the second space 201 have a good sealing effect, making the entire film chip 10 simple in structure and easy to process.

[0056] The second layer 200, culture layer 110, substrate 120, and porous membrane 300 are all translucent to allow observation of the growth status of cells adsorbed on the porous membrane 300. Optionally, the second layer 200, culture layer 110, and substrate 120 can each be independently made of glass, plastic, PDMS (polydimethylsiloxane), etc., and the porous membrane 300 can be a PC (polycarbonate) membrane or a PET (polyethylene terephthalate) membrane, etc. The second layer 200 and culture layer 110, and the culture layer 110 and substrate 120 can be bonded and encapsulated by hot pressing, ultrasound, laser, etc.

[0057] Understandably, the higher the average expansion degree of the porous membrane 300, the more stable the growth state of the cells adsorbed on the surface of the porous membrane 300. Optionally, the edge region of the porous membrane 300 is fixed to at least one of the first layer 100 and the second layer 200. For example, the porous membrane 300 is bonded and encapsulated to the first layer 100 (specifically the culture layer 110) by means of hot pressing, ultrasound, laser, etc.; or, as... Figure 8 As shown, the membrane chip 10 also includes a support ring 400. The edge region of the porous membrane 300 is fixed to the support ring 400. The support ring 400 is installed in at least one of the first layer 100 and the second layer 200. For example, when the first layer 100 has a second space 201, a step is formed at the connection between the first culture chamber 1011 and the second culture chamber 2011. The step has a step surface 113 perpendicular to the direction of gravity G. The support ring 400 is disposed on the step surface 113 to improve the uniformity and stability of the opening of the porous membrane 300.

[0058] like Figure 4 and Figure 5 , Figure 7 and Figure 8 As shown, a first hole 204 is formed on the surface of the second layer 200 opposite to the surface of the first layer 100. The first hole 204 extends toward the first layer 100 and communicates with the second culture chamber 2011, thus allowing substances in the second culture chamber 2011 to directly enter and exit the second culture chamber 2011 through the first hole 204. For example, taking skin epithelial cells as an example, one method of using the membrane chip 10 of this embodiment is described as follows:

[0059] Using a pipette, a cell suspension containing skin epithelial cells is added through the first well 204 into the second culture chamber 2011 above the porous membrane 300. Using a pipette, culture medium is injected through at least one of the first inlet ports 202, filling the entire first culture chamber 1011. After the membrane chip 10 is placed in an incubator and incubated for a period of time, skin epithelial cells are deposited and adsorbed onto the upper surface of the porous membrane 300. Using a pipette, the cell suspension (the remaining liquid after the skin epithelial cells adsorbed onto the surface of the porous membrane 300) in the second culture chamber 2011 is aspirated through the first well 204, exposing the skin epithelial cells adsorbed on the upper surface of the porous membrane 300 to air, simulating the growth environment of normal skin. The membrane chip 10 is placed on a shaker, which oscillates at a specific angle, causing the culture medium in the first culture chamber 1011 to flow back and forth between the multiple first inlets 202. This continuous flow of culture medium through the porous membrane 300 creates a dynamic culture of the skin epithelial cells. By adjusting the drug components and dosage in the culture medium, it is possible to study the effects of drugs on skin epithelial cells.

[0060] In another embodiment, when it is necessary to grow cells on both sides of the porous membrane 300, cell suspension can be injected into the first culture chamber 1011 through the first inlet 202 and into the second culture chamber 2011 through the second inlet 203. After the cells on the corresponding surface of the porous membrane 300 are adsorbed and stabilized each time, the membrane chip 10 is flipped over to pour out the cell suspension (the liquid remaining after the cells are adsorbed on the surface of the porous membrane 300). The corresponding culture medium is then injected into the first culture chamber 1011 and the second culture chamber 2011 to perform cell culture on both sides of the porous membrane 300.

[0061] Since the membrane chip 10 needs to be moved frequently during the culture process, which will accelerate the evaporation of the culture medium, the setting of the first well 204 can be used to reserve space to store more culture medium and prevent the liquid in the second culture chamber 2011 from being evaporated.

[0062] Optionally, such as Figures 7 to 10 As shown, the second layer 200 has multiple evaporation tanks 205 on its surface opposite to the first layer 100. The evaporation tanks 205 are spaced apart from the first space 101, the second space 201, the first sample inlet 202, and the second sample inlet 203. Each evaporation tank 205 stores an anti-evaporation liquid, such as deionized water or PBS buffer. During cell culture, the anti-evaporation liquid in the evaporation tank 205 is exposed to the surface of the membrane chip 10, allowing it to evaporate and carry away heat, thus reducing the evaporation of the culture medium in both the first space 101 and the second space 201.

[0063] The second layer 200 has multiple first inlet ports 202 and multiple second inlet ports 203; at least one evaporation tank 205 is provided between two adjacent first inlet ports 202, between two adjacent second inlet ports 203, and between adjacent first inlet ports 202 and second inlet ports 203, to increase the evaporation area and improve the anti-drying effect.

[0064] The shape of the first culture chamber 1011 is similar to that of the second culture chamber 2011, and can be circular, regular polygonal, or regular polygonal with rounded corners. Preferably, as... Figure 10 As shown, both the first culture chamber 1011 and the second culture chamber 2011 are circular, which facilitates the arrangement of the shape and position of the first sample inlet 202, the second sample inlet 203, the first hole 204 and the evaporation tank 205, making the four components compact and occupying less space. This helps to set up more culture units 11 in a limited space and improve the throughput of the membrane chip 10.

[0065] In the accompanying drawings of this embodiment, the same or similar reference numerals correspond to the same or similar components. In the description of this application, it should be understood that if terms such as "upper," "lower," "left," and "right" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, they are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, the terms used to describe positional relationships in the accompanying drawings are only for illustrative purposes and should not be construed as limiting this patent. For those skilled in the art, the specific meaning of the above terms can be understood according to the specific circumstances.

[0066] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A gravity-driven high-throughput film chip, characterized in that, For mounting on a driving device, and for controlling the driving device to regulate the reciprocating motion state of the gravity-driven high-throughput film chip, the gravity-driven high-throughput film chip includes: The first layer has a first space, which includes a first culture chamber; A porous membrane is provided corresponding to the first culture chamber; and The second layer is stacked on top of the first layer along the direction of gravity, and at least one of the first layer and the second layer has a second space, the second space including a second culture chamber, which is located on the side of the porous membrane away from the first culture chamber in the direction of gravity; the second layer has at least one first injection cell and at least one second injection cell, one end of the first injection cell is connected to the first culture chamber and the other end forms a first injection port on the surface of the second layer away from the first layer, one end of the second injection cell is connected to the second culture chamber and the other end forms a second injection port on the surface of the second layer away from the first layer, and both the first injection port and the second injection port are higher than the second culture chamber in the direction of gravity; The first space further includes a first flow channel communicating with the first injection cell and the first culture chamber, and the second space further includes a second flow channel communicating with the second injection cell and the second culture chamber; in the direction of gravity, the first injection cell and the second injection cell have depth, and the first flow channel is located below the first injection cell, and the second flow channel is located below the second injection cell.

2. The gravity-driven high-throughput film chip according to claim 1, characterized in that, The porous membrane covers the junction between the first culture chamber and the second culture chamber to prevent the substances in the first culture chamber and the second culture chamber from being directly exchanged without the porous membrane.

3. The gravity-driven high-throughput film chip according to claim 1, characterized in that, The first flow channel and the first culture chamber are located in a first plane perpendicular to the direction of gravity; The second flow channel and the second culture chamber are located in a second plane perpendicular to the direction of gravity; Both the first plane and the second plane are parallel to the porous membrane and are located on opposite sides of the porous membrane in the direction of gravity.

4. The gravity-driven high-throughput film chip according to claim 3, characterized in that, The number of the first injection ports is multiple, and the first space includes multiple first flow channels that are equal in number and correspond one-to-one with the number of the first injection ports, and the multiple first flow channels are arranged in a rotationally symmetrical or axisymmetric manner. The number of second injection ports is multiple, and the second space includes multiple second flow channels that are equal in number to and correspond one-to-one with the number of second injection ports, and the multiple second flow channels are arranged rotationally symmetrically or axially symmetrically.

5. The gravity-driven high-throughput film chip according to claim 4, characterized in that, The first space is a rotationally symmetric space or an axisymmetric space; and / or, The second space is a rotationally symmetric space or an axisymmetric space.

6. The gravity-driven high-throughput film chip according to claim 3, characterized in that, The first layer has a first mating surface that connects to the second layer, the first layer has a second space, and the second culture chamber and the second flow channel are formed on the first mating surface.

7. The gravity-driven high-throughput film chip according to claim 6, characterized in that, The first layer includes a culture layer and a base plate stacked in the direction of gravity; the culture layer includes a first mating surface connected to the second layer and a second mating surface facing away from the first mating surface; The first culture chamber and the first flow channel are located on the second docking surface, and the second docking surface is connected to the base plate.

8. The gravity-driven high-throughput film chip according to claim 1, characterized in that, The edge region of the porous membrane is fixed to at least one of the first layer and the second layer; or, The membrane chip further includes a support ring, the edge region of the porous membrane is fixed to the support ring, and the support ring is installed on at least one of the first layer and the second layer.

9. The gravity-driven high-throughput film chip according to claim 1, characterized in that, The second layer has a first hole on its surface opposite to the first layer. The first hole extends toward the first layer and communicates with the second culture chamber, so that the material in the second culture chamber can directly enter and exit the second culture chamber through the first hole.

10. The gravity-driven high-throughput film chip according to claim 1 or 9, characterized in that, The second layer has multiple evaporation tanks on its surface opposite to the first layer; the evaporation tanks are spaced apart from the first space, the second space, the first sample inlet and the second sample inlet. The second layer has a plurality of first inlet ports and a plurality of second inlet ports; at least one of the evaporation tanks is provided between two adjacent first inlet ports, between two adjacent second inlet ports, and between adjacent first inlet ports and second inlet ports.

11. The gravity-driven high-throughput film chip according to claim 1, characterized in that, The shape of the first culture chamber is similar to that of the second culture chamber, and it is a circular hole, a regular polygonal hole, or a regular polygonal rounded corner hole.

12. The gravity-driven high-throughput film chip according to claim 10, characterized in that, The first space, the second space, the first inlet, the second inlet, and the porous membrane form a culture unit. The membrane chip has multiple culture units, and any two culture units are spaced apart.

13. The application of the membrane chip as described in claim 12 in biological model culture and drug analysis.

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