Organ chip

Abstract

The embodiment of the invention provides an organ chip, and relates to the technical field of organ chips, the organ chip comprises at least two function parts and a dielectric layer located between the two adjacent function parts, the at least two function parts are stacked, and each function part is provided with a cell culture chamber; the dielectric layer is used for isolating contact between cells of adjacent cell culture chambers and allowing interaction and substance exchange between signal molecules between the adjacent cell culture chambers, and through the stacked functional part and dielectric layer structure, interaction of the signal molecules is allowed while different cell culture chambers are physically isolated; the air guide part and the connector of the culture box are tightly pressed, the air pressure regulation and control precision and the culture environment stability are guaranteed, the air pressure stability in the culture box is effectively guaranteed, and external microorganisms, dust and other pollutants are effectively prevented from invading the culture environment.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of organ chips, and in particular to an organ chip. BACKGROUND

[0002] With the development of science and technology, organoid and organ chip culture technology can make cells differentiate into cell types with organoid specificity. Through organoid culture technology, cell culture and the organ represented thereby have similar spatial organization and part of the function of the corresponding organ, thereby being able to highly simulate the development of human organs in vitro, and being able to reconstruct the structure of human organs to replace animal models, to provide more possibilities for the research of human diseases.

[0003] At present, three-dimensional culture models are mainly based on two technical routes: one is 3D cell spheres (such as tumor spheres) formed by hanging drop method or low-adhesion plate, and the other is single-layer microfluidic channel based on microfabrication. Although the above methods can partially simulate the microenvironment of tissues, the three-dimensional culture model is mostly 3D cell sphere culture or single-layer channel culture, and the in vitro model cultured thereby cannot realize diversified testing, and the types of the constructed model are relatively single.

[0004] Therefore, there is an urgent need for an organ chip to construct a multi-cell type spatial co-culture system, which can realize diversified testing. SUMMARY

[0005] The embodiment of the present application provides an organ chip, which realizes close compression of the air guide member and the culture box interface, ensures the air pressure regulation accuracy and the culture environment stability, effectively guarantees the air pressure stability in the culture box, and effectively avoids the invasion of external microorganisms, dust and other pollutants into the culture environment.

[0006] The embodiment of the present application provides an organ chip, which comprises: a plurality of culture boxes; a bracket, wherein a plurality of bearing parts for bearing the culture boxes are formed on the bracket; an air guide member, wherein the air guide member is used for filling gas into the culture boxes; and an auxiliary compression mechanism, wherein the auxiliary compression mechanism is arranged at the bottom of the air guide member and corresponds to the culture boxes, and the auxiliary compression mechanism is used for abutting against the culture boxes and positioning the culture boxes when the air guide member is compressed on the top of the culture boxes.

[0007] In a possible implementation, the functional part is provided with a microfluidic channel in communication with the inside of the cell culture chamber, and the microfluidic channel has a liquid inlet hole and a liquid outlet hole.

[0008] In a possible implementation, the functional part is two, and the two functional parts are a first functional part and a second functional part respectively, the first functional part has a first cell culture chamber, and the second functional part has a second cell culture chamber, and the medium layer is located between the first cell culture chamber and the second cell culture chamber and forms a physical isolation.

[0009] In a possible implementation, the microfluidic channel includes a first microfluidic channel arranged in the first functional part and in communication with the first cell culture chamber, the liquid inlet hole includes a first liquid inlet hole arranged in the first functional part, the liquid outlet hole includes a first liquid outlet hole arranged in the first functional part, and the first liquid inlet hole and the first liquid outlet hole are in communication with the first microfluidic channel.

[0010] In a possible implementation, the microfluidic channel includes a second microfluidic channel arranged in the second functional part, the second microfluidic channel is in communication with the second cell culture chamber, the liquid inlet hole includes a second liquid inlet hole arranged in the first functional part, the liquid outlet hole includes a second liquid outlet hole arranged in the first functional part, and the second liquid inlet hole and the second liquid outlet hole are in communication with the second microfluidic channel.

[0011] In a possible implementation, the first microfluidic channel and the second microfluidic channel are projected in the height direction of the functional part in the shape of X.

[0012] In a possible implementation, both sides of the cell culture chamber in the width direction are provided with a negative pressure chamber, a negative pressure hole in communication with the negative pressure chamber is arranged on the functional part, and the negative pressure hole is subjected to suction to make the negative pressure chamber contract and press the cell culture chamber.

[0013] In a possible implementation, the height of the cell culture chamber is greater than or equal to 0.1 mm and less than or equal to 4 mm.

[0014] In a possible implementation, the height of the first cell culture chamber is greater than the height of the second cell culture chamber.

[0015] In a possible implementation, the medium layer is a porous membrane.

[0016] The organ chip provided by the embodiments of the present application realizes the physical isolation of different cell culture chambers while allowing the signal molecules to interact, realizes the close compression of the air guide member and the culture box interface, ensures the air pressure regulation accuracy and the culture environment stability, effectively guarantees the air pressure stability in the culture box, and effectively avoids the invasion of external microorganisms, dust and other pollutants into the culture environment. BRIEF DESCRIPTION OF DRAWINGS

[0017] The drawings incorporated into the specification and forming a part of the specification, show embodiments consistent with the present application, and together with the specification, serve to explain the principles of the present application.

[0018] Figure 1 A top view of an organ chip provided for the present application;

[0019] Figure 2 An exploded structural schematic diagram of an organ chip provided for the present application.

[0020] The figure reference: 100, first functional part; 101, first cell culture chamber; 102, first microfluidic channel; 103, first liquid inlet hole; 104, first liquid outlet hole; 200, second functional part; 201, second cell culture chamber; 202, second microfluidic channel; 203, second liquid inlet hole; 204, second liquid outlet hole; 300, medium layer; 400, negative pressure chamber; 500, negative pressure hole.

[0021] The specific embodiments of the present application have been shown by the above-mentioned drawings, and will be described in more detail hereinafter. These drawings and textual descriptions are not intended to limit the scope of the concept of the present application in any way, but to illustrate the concept of the present application to those skilled in the art by referring to specific embodiments. DETAILED DESCRIPTION

[0022] The exemplary embodiments will be described in detail herein with reference to the accompanying drawings. In the following description, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The implementations described in the following exemplary embodiments are not meant to represent all implementations consistent with the present application. Rather, they are merely examples of apparatus and methods consistent with some aspects of the present application as detailed in the appended claims.

[0023] In the related art, organoid and organ chip culture technology has been able to realize cell differentiation to form specific cell types of organoids, but existing three-dimensional culture models are mostly based on hanging drop method or single-layer microfluidic channel construction. Such models can partially simulate the microenvironment of tissues, but are limited by single culture layer structure and cannot establish a three-dimensional interaction system between multiple cell types. When it is necessary to simulate the signal transmission or material exchange between adjacent organs, the traditional model is difficult to realize the synchronous control of cell spatial isolation and molecular interaction, resulting in limited test scenarios and insufficient model complexity.

[0024] In order to solve the above problems, the researchers found in the development process that the traditional single-layer structure cannot meet the needs of multi-organ co-culture, and then thought about how to construct a three-dimensional culture system with physical isolation but allowing molecular exchange. By analyzing the interaction mode between organs in living organisms, it is realized that it is necessary to establish an isolation interface similar to the basement membrane. After many structure verifications, it is finally determined to use a combination scheme of stacked chambers combined with selective permeable medium, which not only maintains the independent growth space of cells, but also realizes the signal transmission across layers.

[0025] Therefore, the organ chip provided in the application comprises at least two functional parts and a medium layer 300 located between the two adjacent functional parts, the at least two functional parts are arranged in a stacked manner, and each functional part has a cell culture chamber; the medium layer 300 is used to isolate the contact between cells in adjacent cell culture chambers and allow the interaction between signal molecules and the exchange of substances between adjacent cell culture chambers, and through the structure of the stacked functional parts and the medium layer 300, the signal molecules are allowed to interact while the different cell culture chambers are physically isolated, the gas guide member is tightly pressed with the culture box interface, the accuracy of air pressure regulation and the stability of the culture environment are ensured, the air pressure stability in the culture box is effectively guaranteed, and the invasion of external microorganisms, dust and other pollutants into the culture environment is effectively avoided.

[0026] The technical solutions of the application and how the technical solutions of the application solve the above technical problems will be described in detail in specific embodiments. The following specific embodiments can be combined with each other, and the same or similar concepts or processes can not be described again in some embodiments. The embodiments of the application will be described below with reference to the drawings.

[0027] In combination with Figure 1 , Figure 2 , the embodiment of the application provides an organ chip, which comprises at least two functional parts and a medium layer 300 located between the two adjacent functional parts, each functional part has a cell culture chamber, and the medium layer 300 is used to isolate the contact between cells in adjacent chambers while allowing the interaction of signal molecules and the exchange of substances.

[0028] Among them, the functional part is a structural unit carrying the function of cell culture, which can be prepared by microfabrication technology using polydimethylsiloxane material to form a chamber structure with a specific geometric shape. The stacked arrangement refers to the arrangement in the vertical direction. The medium layer 300 is an isolation interface with selective permeability, which can be a porous polymer membrane with controllable pore size, such as a polycarbonate membrane or a polyester membrane, and the pore size range can be set to allow substances with a molecular weight less than 500 Da to pass freely.

[0029] Specifically, the two functional parts form upper and lower culture units in a stacked manner, and each culture unit has an independent cell culture chamber inside. The medium layer 300 is sandwiched between the two adjacent upper and lower cell culture chambers to form a physical isolation barrier. In the culture process, the endothelial cells in the upper chamber and the hepatocytes in the lower chamber grow independently, and the cross-layer diffusion of small molecular substances such as cytokines and metabolites is realized through the porous structure of the medium layer 300. This structure not only avoids the direct contact between different cell types leading to abnormal fusion, but also retains the biochemical signal transmission path similar to the in-vivo inter-organ.

[0030] Compared with the related art, the traditional single-layer microfluidic system can only realize planar flow control, while the stacked structure of the scheme expands the culture space in the vertical direction and allows the establishment of a three-dimensional co-culture system of multiple cell types. Although the cell spheres formed by the existing hanging drop method have a three-dimensional structure, the spatial distribution and interaction interface of different cell populations cannot be accurately controlled. The selective permeation characteristics of the medium layer 300 are used to realize accurate regulation of cell spatial isolation and molecular interaction.

[0031] Through the above technical scheme, the application constructs a three-dimensional culture environment that can simulate the interaction of multiple organs, solving the technical problem that the traditional model cannot simultaneously realize cell physical isolation and molecular interaction. The structure supports the differentiation and growth of different cell types in independent chambers, and at the same time establishes a controllable material exchange channel through the medium layer 300, providing an effective in vitro model for studying the interaction between organs, drug metabolism across the barrier, and other complex physiological processes.

[0032] The application further provides that the functional part is provided with a microfluidic channel in communication with the inside of the cell culture chamber.

[0033] The microfluidic channel is a micro-pipe structure connecting the cell culture chamber and can be made by photolithography or 3D printing technology, which is used to establish a material transmission path between the cell culture chamber and the outside world.

[0034] The liquid inlet hole is a liquid injection opening provided on the surface of the functional part, which can be realized by a conical hole or a funnel-shaped structure, and is used to continuously deliver culture medium or experimental reagents to the microfluidic channel.

[0035] The liquid outlet hole is a liquid discharge opening provided on the surface of the functional part, which can be realized by a cylindrical or tapered structure, and is used to discharge cell metabolites or completed culture medium.

[0036] Specifically, the microfluidic channel forms a communication network with the cell culture chamber. When the culture medium is injected through the liquid inlet hole, the liquid flows through the cell culture chamber along the microfluidic channel, and then is discharged through the liquid outlet hole. The channel system can maintain the continuous supply of nutrients in the cell culture chamber, while avoiding the local accumulation of metabolic waste. The path design of the microfluidic channel can independently control the fluid circulation of different functional parts, so that each cell culture chamber has an independent material exchange path.

[0037] Compared with the related art, the 3D cell spheres formed by the traditional hanging drop method lack a directional material circulation mechanism, and the single-layer microfluidic channel cannot realize independent control of multiple chambers. The scheme integrates the microfluidic channel and the multi-level functional part to construct a directional material circulation system while maintaining cell spatial isolation, solving the technical bottleneck that the traditional model cannot realize co-culture of multiple types of cells.

[0038] Through the technical solution, the dynamic material exchange function of the cell culture chamber is realized, so that cells in different functional parts can be cultured in independent microenvironments, and the cell activity is maintained through controllable fluid circulation. The design effectively avoids the cross contamination problem that may occur in multi-chamber co-culture, and provides an expandable fluid control basis for constructing complex organ models.

[0039] The application further proposes that the functional part is two, and the two functional parts are a first functional part 100 and a second functional part 200 arranged in an upper and lower stack, the first functional part 100 has a first cell culture chamber 101, the second functional part 200 has a second cell culture chamber 201, and the medium layer 300 is located between the first cell culture chamber 101 and the second cell culture chamber 201 and forms a physical isolation.

[0040] Among them, the functional part is a structural unit carrying the cell culture chamber, which can be realized by injection molding of transparent polymer materials, and the stacked arrangement can construct a multi-level cell co-culture space. The cell culture chamber is a three-dimensional culture space for accommodating specific types of cells, which can be realized by forming grooves of different sizes through mold injection. Different chambers can culture different cells such as liver cells and vascular endothelial cells. The medium layer 300 is a selective permeation isolation layer, which can be realized by using a polycarbonate porous membrane with a pore size of 0.4-3 μm, which can prevent cell migration and contact, and allow small molecule substances such as cytokines to cross-layer transmission.

[0041] Specifically, the first functional part 100 and the second functional part 200 form a double-layer structure by vertical stacking, and the first cell culture chamber 101 and the second cell culture chamber 201 are located at different levels. The medium layer 300 is clamped between the two functional parts, and the edges are fixed with the functional parts by hot pressing sealing. During the culture process, liver cells are inoculated in the first cell culture chamber 101, and vascular endothelial cells are inoculated in the second cell culture chamber 201. The porous structure of the medium layer 300 allows metabolic products and signal molecules to diffuse between the chambers, but completely blocks the direct contact of the two types of cells. This spatial isolation design allows different cell types to maintain independent microenvironments, while simulating the interaction between real organs through material exchange.

[0042] Compared with the related art, the traditional single-layer microfluidic chip can only realize planar co-culture and cannot construct a multi-layer culture system with spatial isolation. The present scheme realizes the precise separation of the two cell culture chambers in three-dimensional space through the vertical stacking of the double-layer structure, which not only preserves the independence of the cell types, but also establishes a controllable interaction channel. Compared with the random cell ball distribution formed by the hanging drop method, this structure can accurately control the spatial arrangement relationship of different cells.

[0043] By the technical solution, the application realizes accurate co-culture of two different cells in three-dimensional space, and solves the problem that a single-layer culture system cannot simulate the three-dimensional interaction between organs. Different cell types can form specific tissue structures in independent chambers, and controlled material exchange is performed through the medium layer 300, thereby providing an effective platform for constructing complex physiological models such as liver blood sinuses.

[0044] The application further provides an organ-on-a-chip, which comprises at least two function units arranged in a stack, each function unit having a cell culture chamber, and a medium layer 300 arranged between two adjacent function units to isolate cell contact and allow signal molecule interaction. The function unit is provided with a microfluidic channel in communication with the cell culture chamber, and the microfluidic channel has a liquid inlet hole and a liquid outlet hole. The first function unit 100 is provided with a first microfluidic channel 102 in communication with the first cell culture chamber 101, and the first liquid inlet hole 103 and the first liquid outlet hole 104 are arranged on the first function unit 100 and in communication with the first microfluidic channel 102.

[0045] The microfluidic channel is a micro-pipe structure for guiding the flow of culture solution, which can be formed on a polymer substrate by laser engraving or photolithography process, and the flow direction of the culture solution is controlled to realize the delivery of nutrients. The liquid inlet hole and the liquid outlet hole are opening structures arranged on the surface of the function unit, which can be circular holes with a diameter ranging from 0.1 mm to 0.5 mm, and are used to connect an external liquid supply device to form a circulation path. The first microfluidic channel 102 is an independent flow channel system in direct communication with the first cell culture chamber 101, which can be designed in a serpentine or spiral shape to avoid cross contamination with other flow channels through physical isolation.

[0046] Specifically, the first function unit 100 forms a closed circulation system with the first cell culture chamber 101 through the independently arranged first microfluidic channel 102. When the culture solution is injected through the first liquid inlet hole 103, it flows through the first cell culture chamber 101 along the specific path of the first microfluidic channel 102, and is finally discharged from the first liquid outlet hole 104. This structure enables the first cell culture chamber 101 to maintain an independent fluid environment, while allowing selective material exchange with adjacent chambers through the medium layer 300. For example, when simulating the interaction between liver and intestine, the first chamber can culture liver cells and control the concentration of metabolites through a dedicated flow channel, and the medium layer 300 allows specific molecules such as bile acids to be delivered to the adjacent chamber.

[0047] Compared with the related art, the traditional single-layer microfluidic system cannot realize independent fluid control of different chambers, resulting in cross interference when multiple cells are co-cultured. The scheme configures a dedicated microfluidic channel for each functional part, so that different cell culture chambers can maintain an independent culture environment and also interact under control through the medium layer 300. Compared with the design of sharing flow channels in the related art, this split flow channel structure effectively avoids the problem of culture failure caused by mixing of secretions of different cells.

[0048] Through the above technical scheme, the present application realizes parallel independent control of multiple cell culture chambers, which can accurately adjust the culture solution composition and flow parameters of each chamber while ensuring signal interaction between cells. This solves the technical contradiction between independent culture of multiple tissues and cross-tissue interaction of existing organ chips, and provides a reliable foundation for constructing complex organ interaction models.

[0049] The present application further proposes that the microfluidic channel includes a second microfluidic channel 202 arranged in the second functional part 200, the second microfluidic channel 202 being in communication with the second cell culture chamber 201, the liquid inlet hole includes a second liquid inlet hole 203 arranged in the first functional part 100, the liquid outlet hole includes a second liquid outlet hole 204 arranged in the first functional part 100, and the second liquid inlet hole 203 and the second liquid outlet hole 204 are in communication with the second microfluidic channel 202.

[0050] Among them, the second microfluidic channel 202 is a fluid transport path located in the second functional part 200 and in communication with the second cell culture chamber 201, which can be realized by forming a groove structure on the surface or inside of the second functional part 200 through microprocessing technology, for transporting or discharging culture solution to the second cell culture chamber 201.

[0051] Among them, the second liquid inlet hole 203 and the second liquid outlet hole 204 are openings arranged in the first functional part 100 and in communication with the second microfluidic channel 202, which can be realized by arranging through holes at the corresponding positions of the first functional part 100 and the second microfluidic channel 202, so that the fluid can flow across the layers.

[0052] Specifically, the second microfluidic channel 202 is a fluid path independent of the first microfluidic channel 102, and its liquid inlet hole and liquid outlet hole are integrated in the first functional part 100. When the culture solution is injected through the second liquid inlet hole 203, the fluid enters the second microfluidic channel 202 of the second functional part 200 through the through hole of the first functional part 100, and finally flows into the second cell culture chamber 201; in reverse flow, the culture solution flows back from the second cell culture chamber 201 to the second liquid outlet hole 204 for discharge. Through this cross-layer design, the fluid input and output of the second cell culture chamber 201 do not need to occupy additional space on the outer surface of the second functional part 200, thereby simplifying the fluid control complexity of the multi-layer structure.

[0053] Compared with the related art, the existing single-layer microfluidic channel can only realize fluid circulation of a single chamber, and the scheme can make multi-layer functional parts share the same fluid interface on the same side by integrating the inlet and outlet holes of the second microfluidic channel 202 in the first functional part 100, thereby maintaining the compactness of the multi-layer structure and realizing independent fluid control of different chambers.

[0054] Through the above technical scheme, the present application can realize independent fluid supply and discharge of different functional parts in a multi-layer organ chip, avoid fluid path cross interference during multi-chamber co-culture, and reduce the number of external interfaces, thereby providing an expandable microfluidic architecture basis for constructing a multi-cell type spatial co-culture system.

[0055] The present application further proposes that the first microfluidic channel 102 and the second microfluidic channel 202 are projected in the height direction of the functional part in an X shape.

[0056] The height direction projection of the functional part is the planar distribution form of the microfluidic channel when viewed vertically from the functional part, and the cross layout flow channel structure can be used to realize it. The two microfluidic channels are staggered in space through the X-shaped arrangement.

[0057] The X shape is that the two microfluidic channels extend in a non-parallel and non-vertical manner on the projection plane, and the symmetric or asymmetric cross angle design can be used to realize it. For example, the cross angle range can be controlled between 30° and 150°.

[0058] Specifically, the first microfluidic channel 102 and the second microfluidic channel 202 are respectively arranged in the two functional parts, and are spatially separated in the vertical direction through the X-shaped projection layout. When the fluid is injected from the first inlet hole 103 and the second inlet hole 203 respectively, the X-shaped flow channel structure makes the fluid flow paths in the two channels not interfere with each other, and at the same time realizes signal molecule interaction through the porous membrane of the medium layer 300. Due to the cross characteristic of the X-shaped projection, the inlet and outlet of the two microfluidic channels can be located on different sides of the functional part, thereby simplifying the connection layout of the external fluid control equipment.

[0059] Compared with the related art, the channel of the existing microfluidic chip usually adopts a parallel or straight line type design, which causes the fluid paths of different functional parts to overlap or interfere with each other, affecting the material exchange efficiency. The present application optimizes the space utilization rate between channels while ensuring independent fluid control, thereby reducing the risk of cross contamination.

[0060] Through the technical solution, the application can realize independent transportation and precise regulation of fluids in two functional parts, and promote signal interaction between cells through the medium layer 300, thereby supporting co-culture of multiple types of cells under spatial separation conditions, and meeting the construction requirements of complex in vitro models.

[0061] The application further provides an organ chip, both sides of the cell culture chamber in the width direction are provided with negative pressure chambers 400, and negative pressure holes 500 communicating with the negative pressure chambers 400 are arranged on the functional part. The negative pressure holes 500 are extracted to make the negative pressure chambers 400 contract and press the cell culture chamber.

[0062] The negative pressure chamber 400 is a cavity structure arranged on both sides of the cell culture chamber and capable of forming a negative pressure inside, and specifically can be realized by adopting a sealed cavity made of an elastic material. When the negative pressure hole 500 is extracted, the cavity volume contracts to generate a pressing effect on the cell culture chamber. The negative pressure hole 500 is a through hole connecting the negative pressure chamber 400 and the outside, and specifically can be realized by arranging a through hole on the surface of the functional part. The internal pressure of the negative pressure chamber 400 can be adjusted by connecting the hole to an external air extraction device.

[0063] Specifically, when the negative pressure hole 500 is connected to the air extraction device, the air inside the negative pressure chamber 400 is extracted to form a negative pressure environment, and at this time, the side wall of the negative pressure chamber 400 made of an elastic material collapses and contracts inward. The mechanical force generated by the contraction directly acts on the side wall of the cell culture chamber, so that the internal space of the chamber is periodically or continuously compressed and deformed. The deformation can simulate the mechanical stress borne by the human organ in physiological activities, such as the pressure change generated by intestinal peristalsis or the extrusion effect caused by blood vessel contraction.

[0064] Compared with the related art, the traditional three-dimensional culture model only realizes material exchange through static culture medium diffusion and cannot actively apply mechanical stress stimulation. Although the existing microfluidic chip can realize fluid driving, it lacks precise control ability of the structural deformation of the culture chamber. The negative pressure chamber 400 structure arranged independently realizes dynamic regulation of the spatial form of the cell culture chamber.

[0065] Through the technical solution, the application can provide a controllable mechanical stress environment for cell culture, so that the cultured organoids can more realistically simulate the mechanical stimulation borne by the human organ in a physiological or pathological state. The dynamic extrusion effect can promote mechanical signal transmission between cells and enhance the extracellular matrix reconstruction ability, thereby constructing an in vitro tissue model with physiological relevance.

[0066] The application further provides that the height of the cell culture chamber is greater than or equal to 0.1 mm and less than or equal to 4 mm, for example, the height of the cell culture chamber is 0.1 mm, 3 mm or 4 mm.

[0067] The height of the cell culture chamber is the maximum distance of the chamber in the direction perpendicular to the functional part layering direction, which can be achieved by mold forming or 3D printing process. The height range can provide sufficient space for different cell types to form a three-dimensional structure, while avoiding the reduction of nutrient transport efficiency due to excessive height.

[0068] Specifically, the height of the cell culture chamber is limited to the range of 0.1mm to 4mm, for example, typical values of 0.1, 1.2mm or 3.5mm are adopted. Within this range, the chamber can accommodate a sufficient number of cells to form organoid structures, and the material exchange between adjacent chambers can be achieved through the porous membrane of the medium layer 300. By controlling the upper limit of the height, the problem of uneven distribution of oxygen or metabolites in the chamber can be prevented. By setting the lower limit of the height, the cell layer has a basic three-dimensional growth space, and the functional loss caused by planar culture is avoided.

[0069] Compared with related technologies, the traditional three-dimensional culture model usually adopts single-layer chambers or microfluidic channels with fixed height. For example, the size of the cell ball formed by the hanging drop method is difficult to control accurately, and the height of the single-layer microfluidic channel is usually less than 0.1mm, which limits the cell accumulation. The height range of the present application can dynamically adjust the spatial structure of the chamber of different organ modules according to the cell type. For example, liver cells can use a higher chamber to simulate liver lobule morphology, and vascular endothelial cells can adapt to a lower chamber to form a flattened structure.

[0070] Through the above technical solutions, the present application can provide a three-dimensional space for a multi-cell co-culture system to adapt to different tissue characteristics. Under the premise of maintaining the material exchange efficiency, the controllable growth of the organoid structure is realized, thereby supporting the construction and diversified testing requirements of complex pathological models.

[0071] The present application further proposes that the height of the first cell culture chamber 101 is greater than the height of the second cell culture chamber 201.

[0072] The first cell culture chamber 101 is a three-dimensional culture space for carrying a specific type of cell in the upper layer structure of the organ chip, which can be achieved by adjusting the mold size or the height of the layer structure. The height difference can provide differentiated growth space for different cell types.

[0073] The second cell culture chamber 201 is a three-dimensional culture space for carrying another type of cell in the lower layer structure of the organ chip, which can be achieved by independently designing the chamber height or using an adjustable support structure. The height difference helps to simulate the physiological microenvironment of different organs.

[0074] Specifically, by making the height of the first cell culture chamber 101 greater than the height of the second cell culture chamber 201, a multi-layer culture system with different vertical spatial distribution can be formed in the same organ chip. For example, the first cell culture chamber 101 can accommodate cell types that require more growth space, while the second cell culture chamber 201 is adapted to cell types that require close arrangement, both of which achieve signal molecule interaction through the medium layer 300. This height difference further optimizes the culture conditions of different cell types, enabling the co-culture system to more realistically simulate the interaction between organs in vivo.

[0075] In some embodiments, the height of the first cell culture chamber 101 can be set close to the upper limit, and the height of the second cell culture chamber 201 is set close to the lower limit, thereby maximizing the culture compatibility of different cell types in a limited space.

[0076] Compared with related art, traditional organ chips usually use uniform height culture chambers, which limits the spatial distribution of different cell types and cannot effectively simulate the three-dimensional interaction between organs. The present scheme breaks through the limitation of single spatial structure by differentiating the chamber height, enabling the multi-cell co-culture system to more flexibly adapt to the physiological needs of different organs.

[0077] Through the above technical scheme, the present application can provide a differentiated three-dimensional growth environment for a variety of cell types, enhance the simulation capability of the organ chip for complex physiological systems, and at the same time avoid the problem of culture failure caused by space competition of different cells.

[0078] The present application further proposes that the medium layer 300 is a porous membrane.

[0079] The porous membrane is a thin film material with through-hole structure, which can be made of polycarbonate or polyester material, and the pore diameter can be controlled within the range of 0.1 microns to 10 microns. This structure allows signal molecules and nutrients with a molecular weight below a certain threshold to diffuse through the pores, while blocking cell migration to achieve physical isolation.

[0080] Specifically, the porous membrane is arranged between the cell culture chambers of adjacent functional parts, and its through-hole structure forms a material exchange channel. When cells of different functional parts are cultured in their respective chambers, small molecule substances such as growth factors and metabolic products secreted by the cells can be transferred across layers through the pores, while the cells themselves are constrained in the independent chambers due to size limitations. This structure enables multiple cell types to interact under spatial isolation conditions, for example, hepatocytes and vascular endothelial cells can transfer albumin or cytokines through the porous membrane to simulate the interaction between organs.

[0081] Compared with related technologies, the conventional three-dimensional culture model usually adopts a solid isolation layer or a single fluid channel, and cannot simultaneously realize physical isolation and material exchange. For example, some devices use non-permeable polymer membranes to completely block the contact between cells, resulting in a loss of physiological relevance in the co-culture system. A porous membrane maintains an independent cell growth environment and retains the biological molecule exchange path through a selective permeation mechanism.

[0082] Through the above technical solutions, the application solves the problem that the conventional organ chip cannot realize spatial co-culture of multiple cell types. The controllable permeability provided by the porous membrane enables cells in different chambers to establish a biomimetic microenvironment. For example, in drug toxicity testing, liver cell metabolites can act on downstream myocardial cells through pore action, thereby completely simulating in vivo organ cascade reactions. This structure significantly improves the biological relevance of in vitro models and provides an effective platform for complex pathological mechanism research and multi-organ interaction analysis.

[0083] Finally, it should be noted that: other embodiments of the application will be readily apparent to those skilled in the art upon considering the description and practicing the application disclosed herein. The application is intended to cover any variations, uses, or adaptive changes of the application that follow the general principles of the application and include common knowledge or conventional techniques in the art that are not disclosed by the application, and is not limited to the precise structure described above and shown in the drawings, and various modifications and changes can be made without departing from the scope thereof. The scope of the application is only limited by the appended claims.

Claims

1. An organ-on-a-chip, characterized in that, include: At least two functional parts are stacked together, and each of the functional parts has a cell culture chamber; A medium layer is located between two adjacent functional units. The medium layer is used to isolate cell-to-cell contact between adjacent cell culture chambers and to allow interaction and exchange of substances between signal molecules between adjacent cell culture chambers.

2. An organ-on-a-chip according to claim 1, characterized in that: The functional part is provided with a microfluidic channel communicating with the interior of the cell culture chamber, and the microfluidic channel has an inlet hole and an outlet hole.

3. An organ-on-a-chip according to claim 2, characterized in that: The functional part consists of two parts, namely a first functional part and a second functional part. The first functional part has a first cell culture chamber, and the second functional part has a second cell culture chamber. The medium layer is located between the first cell culture chamber and the second cell culture chamber and forms a physical isolation.

4. An organ-on-a-chip according to claim 3, characterized in that: The microfluidic channel includes a first microfluidic channel disposed in the first functional part and communicating with the first cell culture chamber. The liquid inlet includes a first liquid inlet disposed in the first functional part. The liquid outlet includes a first liquid outlet disposed in the first functional part. The first liquid inlet and the first liquid outlet are communicating with the first microfluidic channel.

5. An organ-on-a-chip according to claim 4, characterized in that: The microfluidic channel includes a second microfluidic channel disposed in the second functional part, the second microfluidic channel being connected to the second cell culture chamber, the liquid inlet includes a second liquid inlet disposed in the first functional part, the liquid outlet includes a second liquid outlet disposed in the first functional part, and the second liquid inlet and the second liquid outlet are connected to the second microfluidic channel.

6. An organ-on-a-chip according to claim 5, characterized in that: The first and second microfluidic channels are projected in an X-shape along the height of the functional part.

7. An organ-on-a-chip according to any one of claims 1-6, characterized in that: Negative pressure chambers are provided on both sides of the cell culture chamber in the width direction. A negative pressure hole is provided on the functional part to communicate with the negative pressure chamber. When air is drawn from the negative pressure hole, the negative pressure chamber contracts and squeezes the cell culture chamber.

8. An organ-on-a-chip according to any one of claims 1-6, characterized in that: The height of the cell culture chamber is greater than or equal to 0.1 mm and less than or equal to 4 mm.

9. An organ-on-a-chip according to claim 8, characterized in that: The height of the first cell culture chamber is greater than the height of the second cell culture chamber.

10. An organ-on-a-chip according to any one of claims 1-6, characterized in that: The medium layer is a porous membrane.

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