Modularized intelligent breast cancer organ chip system and use method thereof

Abstract

The invention discloses a modular intelligent breast cancer organ chip system and a use method thereof, and aims to simulate the processes of breast cancer primary lesion invasion, lymphatic / hematogenous metastasis and far-end colonization and realize tumor metastasis mechanism research, microenvironment dynamic monitoring and related drug evaluation. All the modules (such as a tumor primary focus module, a bionic blood vessel module, a lymphatic network module and a far-end organ module) have independent cell culture, fluid perfusion control and integrated sensor arrays so as to realize micro-environment dynamic monitoring. The modules are connected and subjected to material exchange through standardized reconfigurable interfaces (including direct fluid butt joint and selective permeable membrane interfaces). The modular architecture endows the system with high flexibility and expandability, can quickly customize and combine specific transfer mechanism research, microenvironment parameter monitoring or drug efficiency evaluation, and ensures the quality control and function specificity of each unit. And a foundation is laid for developing an intelligent sensing and feedback regulation organ chip system for cancer research.

Description

Technical Field

[0001] The present invention belongs to the technical field of bionic organ chips, and specifically relates to a modular intelligent breast cancer organ chip system and a method for using the same. Background Art

[0002] Breast cancer is one of the most common malignancies in women worldwide, and its mortality rate is primarily attributed to tumor metastasis. Breast cancer cells can metastasize to distant organs such as the lungs, bones, and liver via the hematogenous and lymphatic pathways, forming secondary lesions. Despite advances in cancer treatment strategies in recent years, the biological mechanisms of tumor metastasis are complex and dynamic, and existing in vitro research models still struggle to accurately reproduce the spatial structure and microenvironmental changes throughout the metastatic process, limiting our in-depth understanding of its mechanisms and the efficiency of targeted drug development.

[0003] Traditional two-dimensional cell culture models struggle to simulate the three-dimensional structure, biophysical and mechanical environment, and intercellular interactions of tumor tissue. While animal models provide a physiological context to some extent, they suffer from significant interspecies variability, long experimental cycles, poor controllability, and high ethical costs, making them difficult to meet the demands of high-throughput, quantification, and mechanistic analysis. In recent years, organ-on-a-chip technology, a next-generation microphysiological system platform, has garnered widespread attention in tumor metastasis research due to its advantages, including controllable microscale structures, reproducible physiological environments, and multifactor integration.

[0004] However, most existing tumor-related organ-on-chips are custom-designed for a single scenario, with fixed structures, low integration of functional modules, and a lack of standardized interfaces. This hinders multi-module combination and multidimensional mechanism reconstruction. For example, Humayun et al. designed a PDMS-based chip platform as a human organotypic vascularized microfluidic model to study breast cancer cell egress. This platform allows for the study of interactions between cancer and blood vessels and the role of secreted factors in breast cancer cell egress (Humayun M, Ayuso JM, Brenneke RA, et al. Elucidating cancer-vascular paracrine signaling using a human organotypic breast cancer cell extravasation model [J]. Biomaterials, 2021, 270: 120640). This model has the potential to provide valuable insights into how cancer-vascular interactions enable cancer cell egress and lead to the evaluation of therapeutic approaches to prevent cancer cell egress. However, breast cancer metastasis involves the synergistic effects of both hematogenous and lymphatic pathways, and this model simulates a single metastatic pathway in isolation, thus incompletely understanding the metastatic cascade. At the same time, existing chip systems have limited capabilities in real-time monitoring and intelligent feedback regulation. It is difficult to dynamically capture the behavioral changes of cancer cells in different metastasis stages, and it is also difficult to simulate the complex mechanical disturbances, chemical gradient changes, and immune cell synergy in the tumor microenvironment and other multi-factor coupling processes.

[0005] Therefore, there is an urgent need to develop a breast cancer organ-on-chip system with standardized, modular, and intelligent characteristics. The present invention can flexibly construct metastatic microenvironments under various physiological and pathological conditions through a "Lego-style" modular assembly method, support multi-module integration and rapid replacement, and adapt to various sensor arrays to achieve online monitoring and closed-loop control of the microenvironment, thereby providing a more efficient and repeatable in vitro model platform for breast cancer metastasis mechanism research, personalized drug evaluation, and precision treatment. Summary of the Invention

[0006] The present invention aims to address the shortcomings of existing technologies in tumor metastasis mechanism research, in vitro biomimetic organ model construction, and intelligent response capabilities by providing a modular intelligent breast cancer organ-on-a-chip system and its construction method. By incorporating standardized microfluidic channel units, detachable interface components, functional partitioning modules, and sensor integration mechanisms, the present invention enables rapid assembly, flexible reconfiguration, and intelligent control of the organ-on-a-chip system. This system is suitable for modeling the multi-stage metastasis process of breast cancer, including primary lesion invasion, hematogenous / lymphatic transport, and distal colonization, as well as for testing and evaluating anti-metastatic drugs.

[0007] The object of the present invention is achieved through the following technical solutions:

[0008] A modular intelligent breast cancer organ-on-a-chip system, comprising: a standard module channel unit chip layer and a standard chip channel interface component, wherein the two can be freely combined, and different standard module channel unit chip layers, or a standard module channel unit chip layer and its mirror-symmetrical standard module channel unit chip layer, can be assembled in series or in parallel via the standard chip channel interface component to form the modular intelligent breast cancer organ-on-a-chip system;

[0009] The standard module channel unit chip layer is used to realize functions such as cell culture, fluid transport, and material exchange, and includes a microchannel inlet connection, a microchannel chamber, and a microchannel outlet connection; the microchannel inlet connection and the microchannel outlet connection are respectively connected to the two ends of the microchannel chamber;

[0010] The standard chip channel interface component is used to achieve fluid connection and rapid assembly between different standard module channel unit chip layers, including a transition microchannel and protruding microchannel socket metal connecting pipes located on both sides of the transition microchannel and perpendicular to the two ends of the plane where the transition microchannel is located.

[0011] Furthermore, the microchannel socket metal connecting tube is respectively inserted into the microchannel inlet connection and the microchannel outlet connection by interference fit; thereby achieving standardized docking and fluid penetration between different standard module channel unit chip layers, and is used to achieve interference fit connection with the channel socket metal connecting tube in the standard chip channel interface assembly.

[0012] Furthermore, when the standard module channel unit chip layer and its mirror-symmetrical standard module channel unit chip layer are closely combined in parallel with equal sides, the center distance between the microchannel inlet connection of the standard module channel unit chip layer and the microchannel outlet connection of the mirror-symmetrical standard module channel unit chip layer is equal to the center distance between the two microchannel socket metal connecting tubes;

[0013] When two identical standard module channel unit chip layers are relative to each other and bonded up and down, the distance between the centers of the microchannel inlet connections on the same side of the standard module channel unit chip layer is equal to the distance between the centers of the two microchannel socket metal connecting tubes, and the distance between the centers of the microchannel outlet connections on the same side is equal to the distance between the centers of the two microchannel socket metal connecting tubes.

[0014] Furthermore, the front and rear ends of the microchannel chamber are respectively connected to the microchannel inlet connection and the microchannel outlet connection through transition channels; the microchannel inlet connection and the microchannel outlet connection are standardized circular reserved structures; the cross-section of the transition microchannel is a regular rectangle.

[0015] Furthermore, a microsensor array is embedded in the standard module channel unit chip layer to realize the intelligent response capability of the modular intelligent breast cancer organ chip system; the microsensor array is one or more of a micro-pressure sensor, a flow rate sensor, a pH sensor, an oxygen concentration sensor, an electrical impedance sensor or a membrane displacement sensor; the microsensor array is arranged inside the microchannel chamber or on its surface, connected to the standard signal interface led out from the edge of the standard module channel unit chip layer, and connected to the external control system; through real-time data acquisition of the microsensor array, the system can sense the local fluid environment, cell metabolic state or membrane deformation in the standard module channel unit chip layer, and realize automatic adjustment of flow rate, pressure and chemical factor concentration parameters through linkage with a micropump, a pressure drive device or an environmental control module.

[0016] Furthermore, the standard module channel unit chip layer has a length of 6 cm, a width of 3 cm, and a thickness of 0.4 cm; the central microchannel chamber has a rectangular cross-sectional structure with a width of 2 mm, a length of 2 cm, and a height range of 50 to 200 μm; the radius of the standardized circular reserved structure at the microchannel inlet connection and the microchannel outlet connection is 500 μm; the transition channel width at the front and rear ends of the microchannel chamber is 500 μm.

[0017] Furthermore, the standard chip channel interface component has a length of 2.2 cm, a width of 0.5 cm, and a thickness of 0.4 cm; the cross-section of the transition microchannel is a regular rectangle with a side length of 500 μm; the microchannel socket metal connecting tubes are both metal conduits with an inner diameter of 1000 μm, and the center distance between the two microchannel socket metal connecting tubes is 1.5 cm.

[0018] A modular intelligent breast cancer organ-on-a-chip system, comprising a combination of the following:

[0019] Method 1: A fluidic direct-connection docking series model based on a combination of at least two different standard modular channel unit chip layers, comprising: standard modular channel unit chip layers connected in series; the microchannel outlet connections and / or microchannel inlet connections of the two standard modular channel unit chip layers are interference-fitted through the microchannel socket metal connecting tubes of the standard chip channel interface assembly; each of the standard modular channel unit chip layers can achieve independent cell culture, perfusion control, and functional loading, and can be flexibly configured into different functional modules such as a primary tumor lesion module, a vascular module, and a lymphatic module according to research needs;

[0020] Method 2: A linear chemotactic gradient induction model for substance concentration based on a combination of at least two different standard module channel unit chip layers, comprising: an upper standard module channel unit chip layer and a lower standard module channel unit chip layer; the microchannel inlet connection of the upper layer and the microchannel inlet connection of the lower layer respectively serve as injection ports for two solutions of different concentrations, and the injected fluids are brought into parallel flow contact at the front end through a parallel microchannel chamber of equal width and height; because the fluids in the microchannel chambers are in a low Reynolds number laminar flow state, the two solutions are gradually mixed by molecular diffusion at the interface of the microchannel chambers, forming a stable linear concentration gradient in the axial direction of the channel;

[0021] Method 3: A dynamic deformation pressure chamber model based on a combination of at least two different standard module channel unit chip layers, comprising: adding an intermediate flexible film layer on the basis of Method 2, wherein the intermediate flexible film layer is arranged between the upper standard module channel unit chip layer and the lower standard module channel unit chip layer, and the upper standard module channel unit chip layer and the lower standard module channel unit chip layer are sequentially bonded after plasma treatment to form a three-layer closed structure, wherein the microchannel chamber of the upper standard module channel unit chip layer and the microchannel chamber of the lower standard module channel unit chip layer are completely isolated after relative bonding; the upper standard module channel unit chip layer The microchannel chamber serves as a pressure chamber and is connected to an external micro air pump via a microfluidic pipeline. It can periodically apply positive or negative pressure according to a preset program, causing the intermediate flexible film layer to bulge or sink into the microchannel chamber of the lower standard module channel unit chip layer below. The intermediate flexible film layer vibrates back and forth to simulate the dynamic effects of vascular flow or lymphatic vessel rhythm on the luminal morphology and fluid shear force. The microchannel chamber of the lower standard module channel unit chip layer is used to culture cells or drain fluids, and the deformation of the intermediate flexible film layer and the fluid dynamics parameters in the channel are monitored in real time via micro displacement sensors and flow rate sensors integrated into the intermediate flexible film layer or the channel wall.

[0022] Method 4: Freely combine Method 1, Method 2 and Method 3.

[0023] A method for using a modular intelligent breast cancer organ-on-a-chip system, the specific process is as follows:

[0024] The tumor cell invasion-culture-transport model based on the combination of at least three different standard module channel unit chip layers includes: a standard module channel unit chip layer for simulating the primary tumor lesion is provided on one side, and a three-layer structure is formed by the lower blood vessel-co-culture module, the middle porous film layer and the upper distal organ bionic module stacked up and down on the other side; the lower blood vessel-co-culture module and the upper distal organ bionic module are both standard module channel unit chip layers; the porous film layer sandwiched in the middle is used to simulate the process of cancer cells crossing the epithelial barrier, which is an important interface connecting the two functional areas; the primary tumor lesion module is connected to the blood vessel-co-culture module through a microchannel socket metal connecting tube, forming a continuous fluid path for the release of front-end tumor cells to the middle blood vessel barrier path; the vascular-co-culture module and the distal organ module are directly bonded through the middle porous film layer to form a vertical migration system with upper and lower dual channels; breast cancer cells are seeded in the primary tumor lesion module to simulate their growth, spread or shedding process in situ; the microchannel chamber of the vascular-co-culture module can be paved with vascular endothelial cells, and immune cells can be introduced when necessary to construct a microenvironment barrier that is closer to the real pathological state; the middle porous film layer can use a porous polymer membrane or hydrogel membrane with a specific pore size and permeability to form a simulated interface with a physiological barrier function, which is used to restore the behavior of cancer cells crossing the endothelial structure during invasion; the distal organ bionic module is used to capture migrated cancer cells and reproduce their colonization and spread environment in distal tissues.

[0025] Furthermore, the blood vessel-co-culture module can be replaced with a lymphatic network module to simulate the invasion, entry and migration behavior of breast cancer cells in the lymphatic transport pathway, thereby realizing the reconstruction and dynamic monitoring of the tumor cell-lymphatic system metastasis mechanism; alternatively, the blood vessel-co-culture module is connected in series with a standard module channel unit chip layer for simulating the lymphatic pathway through a standard chip channel interface component, thereby sequentially including four different functional areas in one system: the tumor primary lesion module, the blood vessel module, the lymphatic network module and the distal organ module, constructing a multi-stage tumor metastasis pathway model covering both the blood and lymphatic pathways, and realizing dynamic monitoring and intervention research of the microenvironment under multi-channel, multi-interface and multi-factor coupling conditions.

[0026] The present invention provides at least the following beneficial effects:

[0027] Modular Design: This invention utilizes standardized channel units and unified interface components. Functional modules can be freely combined and quickly assembled as needed, enabling serial simulation of multiple physiological stages, including primary tumor sites, vascular / lymphatic transport, and distal colonization. Compared to traditional integrated chip structures, this significantly improves the adaptability of the model and the scalability of experiments. Furthermore, the modular structure is standardized and simple to manufacture, with quick assembly and disassembly, facilitating mass production and repeatable experiments. Standard interfaces ensure connection sealing and fluid continuity, effectively reducing experimental errors and improving system stability and data consistency.

[0028] Scalability: Suitable for a variety of breast cancer metastasis research scenarios. By replacing module combinations, various classic or complex metastasis research platforms can be quickly constructed, including vascular-lymphatic tandem models, mechanical deformation models, linear chemotactic gradient models, and immune cell collaborative invasion models, meeting the experimental needs of different research stages and targeting strategies.

[0029] Intelligent Design: This invention relies on intelligent sensor integration to achieve dynamic monitoring and feedback control. The system integrates a microsensor array for real-time monitoring of key microenvironmental parameters such as pressure, flow rate, pH, oxygen concentration, and cell impedance. It can also communicate with an external control system to automatically adjust perfusion rate, drug release, and mechanical stimulation, providing intelligent feedback capabilities for "self-sensing and self-regulation." BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Schematic diagram of the modular channel unit and standardized interface of a breast cancer organ chip in one embodiment of the present invention;

[0031] Figure 2 A fluid direct connection and docking series model based on a combination of at least two different modular channel units in one embodiment of the present invention;

[0032] Figure 3 A linear chemotaxis gradient induction model of substance concentration based on a combination of at least two different modular channel units in one embodiment of the present invention;

[0033] Figure 4 A dynamically deformable pressure chamber model based on a combination of at least two different modular channel units in one embodiment of the present invention;

[0034] Figure 5 A tumor cell invasion-culture-transport model based on a combination of at least three different modular channel units in one embodiment of the present invention;

[0035] In the figure: 1 standard module channel unit chip layer; 2 standard chip channel interface assembly; 1-1 microchannel inlet connection; 1-2 microchannel chamber; 1-3 microchannel outlet connection; 2-1 transition microchannel; 2-2 microchannel socket metal connection tube A; 2-3 microchannel socket metal connection tube B;

[0036] 3 standard module channel unit chip layer A; 3-1 microchannel inlet connection A; 3-2 microchannel chamber A; 3-3 microchannel outlet connection A;

[0037] 4 standard module channel unit chip layer B; 4-1 microchannel inlet connection B; 4-2 microchannel chamber B; 4-3 microchannel outlet connection B;

[0038] 5 standard module channel unit chip layer C; 5-1 microchannel inlet connection C; 5-2 microchannel chamber C; 5-3 microchannel outlet connection C;

[0039] 6 standard module channel unit chip layer D; 6-1 microchannel inlet connection D; 6-2 microchannel chamber D; 6-3 microchannel outlet connection D;

[0040] 7 standard module channel unit chip layer E; 7-1 microchannel inlet connection E; 7-2 microchannel chamber E; 7-3 microchannel outlet connection E;

[0041] 9 standard module channel unit chip layer F; 9-1 microchannel inlet connection F; 9-2 microchannel chamber F; 9-3 microchannel outlet connection F;

[0042] 10 standard module channel unit chip layer G; 10-1 microchannel inlet connection G; 10-2 microchannel chamber G; 10-3 microchannel outlet connection G;

[0043] 11 standard module channel unit chip layer H; 11-1 microchannel inlet connection H; 11-2 microchannel chamber H; 11-3 microchannel outlet connection H;

[0044] 13 standard module channel unit chip layer I; 13-1 microchannel inlet connection I; 13-2 microchannel chamber I; 13-3 microchannel outlet connection I;

[0045] 8. Middle flexible film layer;

[0046] 12Porous film structure. DETAILED DESCRIPTION

[0047] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0048] like Figure 1As shown, the modular intelligent breast cancer organ chip system provided by the present invention includes: a standard module channel unit chip layer 1 and a standard chip channel interface component 2, which can be freely combined. Different standard module channel unit chip layers 1 can be assembled in series or in parallel through the standard chip channel interface component 2 to form the modular intelligent breast cancer organ chip system. The standard module channel unit chip layer 1 is prepared using a standard soft lithography micromachining process, and can also be designed with mirror symmetry along the central channel axis. Its main structure includes a microchannel inlet connection 1-1, a microchannel chamber 1-2, and a microchannel outlet connection 1-3; the microchannel inlet connection 1-1, the microchannel chamber 1-2, and the microchannel outlet connection 1-3 are open on the upper surface of the standard module channel unit chip layer 1. The microchannel inlet connection 1-1 and the microchannel outlet connection 1-3 are respectively connected to the two ends of the microchannel chamber 1-2. The standard module channel unit chip layer 1 serves as the basic channel module of the present invention and is used to realize functions such as cell culture, fluid transport, and material exchange.

[0049] The standard chip channel interface assembly 2 is used to achieve fluid connection and rapid assembly between various standard module channel unit chip layers. It primarily comprises a transition microchannel 2-1 embedded within the standard chip channel interface assembly 2, and protruding microchannel socket metal connectors A2-2 and B2-3, located on either side of the transition microchannel 2-1 and perpendicular to the plane of the transition microchannel 2-1. These microchannel socket metal connectors A2-2 and B2-3, respectively, are inserted into the microchannel inlet connection 1-1 and microchannel outlet connection 1-3 through an interference fit, thereby achieving standardized docking and fluid communication between different standard module channel unit chip layers. This structure offers plug-and-play functionality and easy assembly and disassembly, facilitating rapid module replacement and functional reconfiguration.

[0050] The modular channel unit chip layer 1 may be embedded with a microsensor array (not shown in the figure) to realize the intelligent response capability of the modular intelligent breast cancer organ chip system. The microsensor array may include, but is not limited to, micro-pressure sensors, flow rate sensors, pH sensors, oxygen concentration sensors, electrical impedance sensors, membrane displacement sensors, etc. The above-mentioned microsensor array can be arranged inside or on the surface of the microchannel chamber 1-2, connected to the standard signal interface led out from the edge of the standard module channel unit chip layer, and connected to the external control system. Through real-time data acquisition of the microsensor array, the system can sense the local fluid environment, cell metabolic state or membrane deformation within the standard module channel unit chip layer, and realize automatic adjustment of parameters such as flow rate, pressure, and chemical factor concentration by linkage with a micropump, pressure drive device or environmental control module.

[0051] In one embodiment, the standard modular channel unit chip layer 1 is a microfluidic chip measuring 6 cm x 3 cm with a thickness of 0.4 cm. A through-hole microchannel structure is incorporated within the standard modular channel unit chip layer 1. The microchannel has a machinable height range of 50 to 200 μm and can be customized based on specific experimental requirements. The microchannel inlet connection 1-1 and microchannel outlet connection 1-3 are standardized circular pre-set structures with a radius of 500 μm, designed to achieve an interference fit connection with the microchannel socket metal connection tubes A2-2 and B2-3 in the standard chip channel interface assembly 2. The central microchannel chamber 1-2 has a rectangular cross-section with a width of 2 mm and a length of 2 cm. The front and rear ends of the microchannel chamber 1-2 are connected to the microchannel inlet connection 1-1 and microchannel outlet connection 1-3, respectively, via transition channels with a width of 500 μm.

[0052] In one embodiment, the standard chip channel interface assembly 2 is a microfluidic chip measuring 2.2 cm × 0.5 cm and 0.4 cm thick. A through-hole microchannel structure is provided within the standard chip channel interface assembly 2, with a rectangular cross-section of 500 μm. The microchannel socket metal connecting tube A2-2 and the microchannel socket metal connecting tube B2-3 are both metal conduits with an inner diameter of 1000 μm, and the center-to-center distance between the two interfaces is 1.5 cm.

[0053] In one embodiment, when the standard module channel unit chip layer 1 and its mirror image mold are closely assembled parallel to the long side of the chip, the center distance between two adjacent microchannel outlet connections and microchannel inlet connections is 1.5 cm. Therefore, the standard chip channel interface component 2 can be directly used for interference fit to achieve fluid series connection of the two standard module channel unit chip layers 1. Furthermore, when two identical standard module channel unit chip layers 1 are placed opposite each other and attached up and down, the center distance between the microchannel outlet connection and the microchannel inlet connection on the same side of the chip short side is also 1.5 cm. The standard interface component 2 can also be used to complete the fluid docking between the modules in the vertical direction. This structure not only ensures the flexibility of module combination, but also takes into account the consistency of interface dimensions under different assembly methods, realizing multi-level and multi-angle fluid interconnection and reconstruction.

[0054] The above design fully considers the characteristics of modularity and intelligence to adapt to the construction of bionic organ-on-a-chip models for different breast cancer research needs. The following are several specific implementation steps:

[0055] like Figure 2In one embodiment, the present invention provides a fluid direct connection and docking series model based on a combination of at least two different modular channel units, including: a standard module channel unit chip layer A3 and a standard module channel unit chip layer B4 connected in series. The microchannel outlet connection A3-3 of the standard module channel unit chip layer A3 and the microchannel inlet connection B4-1 of the standard module channel unit chip layer B4 are interference fit connected through the protruding metal connecting tube assembly of the interface assembly 2. Each of the standard module channel units can realize independent cell culture, perfusion control and functional loading, and can be flexibly configured into different functional modules such as tumor primary lesion module, blood vessel module, lymphatic module, etc. according to research needs. In this embodiment, the standard module channel unit chip layer A3 can be used to simulate the primary lesion or blood vessel port of breast cancer, and the standard module channel unit chip layer B4 can be used as a downstream lymphatic diversion module. The combination of the two is used to simulate the continuous metastasis process of cancer cells after escaping from the blood vessel end and entering the lymphatic system.

[0056] like Figure 3 In one embodiment, the present invention provides a substance concentration linear chemotaxis gradient induction model based on a combination of at least two different modular channel units, comprising: an upper standard modular channel unit chip layer C5 and a lower standard modular channel unit chip layer D6. The two chip layers are directly bonded by surface plasma treatment to form a closed microfluidic channel system that runs through from top to bottom. The microchannel inlet connection C5-1 of the upper channel and the microchannel inlet connection D6-1 of the lower channel serve as injection ports for two solutions of different concentrations, respectively. The injected fluids are brought into parallel contact at the front end through a parallel microchannel chamber C5-2 of the same width and height with the microchannel chamber D6-2. Since the fluid in the channel is in a low Reynolds number laminar flow state, the two solutions are gradually mixed by molecular diffusion at the channel interface, forming a stable linear concentration gradient in the axial direction of the channel. The parallel microchannel chamber C5-2 and the microchannel chamber D6-2 both have rectangular cross-sectional structures, and their width, length and height are the same as those of the standard modular channel unit.

[0057] like Figure 4In one embodiment, the present invention provides a dynamically deformable pressure chamber model based on a combination of at least two different modular channel units, comprising: an upper standard modular channel unit chip layer E7, an intermediate flexible film layer 8, and a lower standard modular channel unit chip layer F9. The standard modular channel unit chip layer E7, intermediate flexible film layer 8, and standard modular channel unit chip layer F9 are plasma-treated and sequentially bonded to form a three-layer closed structure. The upper channel microchannel chamber E7-2 is completely isolated from the lower channel microchannel chamber F9-2 after bonding, with the intermediate flexible film layer 8 positioned between them. The upper channel microchannel chamber E7-2 serves as a pressure chamber, connected to an external micro-air pump via microfluidic piping. It can periodically apply positive or negative pressure according to a preset program, causing the intermediate flexible film layer 8 to bulge or recess into the lower channel microchannel chamber F9-2. The middle flexible film layer 8 is preferably made of a biocompatible elastic material (such as PDMS or hydrogel) with a thickness of approximately 20–50 μm. It vibrates back and forth to simulate the dynamic effects of physiological heartbeats or lymphatic vascular rhythms on luminal morphology and fluid shear forces. The microchannel chamber F9-2 in the lower channel can be used for culturing cells or draining fluids. Micro-displacement sensors and flow rate sensors integrated into the middle flexible film layer 8 or the channel wall provide real-time monitoring of film deformation and fluid dynamics within the channel.

[0058] like Figure 5As shown, in one embodiment, the present invention provides a tumor cell invasion-culture-transport model based on a combination of at least three different modular channel units. The model includes: a standard module channel unit chip layer G10 on one side, serving as a primary tumor lesion module, and a three-layer structure on the other side, formed by stacking a standard module channel unit chip layer I13, a porous film structure 12, and a standard module channel unit chip layer H11. Specifically, the standard module channel unit chip layer I13 is located at the bottom, serving as a blood vessel-co-culture module; the standard module channel unit chip layer 11 is located at the top, serving as a distal organ biomimetic module; and the film layer 12 sandwiched in between is used to simulate the process of cancer cells crossing the epithelial barrier. The standard module channel unit chip layer G10 is connected to the standard module channel unit chip layer I13 via a standard chip channel interface component 2, forming a continuous fluid pathway from the primary lesion module to the co-culture module; the standard module channel unit chip layer I13 and the standard module channel unit chip layer H11 are directly bonded via the intermediate porous film layer 12, thereby forming a double-channel structure. Breast cancer cells can be seeded in the standard module channel unit chip layer G10 to simulate the growth and shedding process of cancer cells in the primary tumor lesion; the upper channel within the standard module channel unit chip layer I13 can be laid with vascular endothelial cells, or immune cells can be further introduced for co-culture with them to construct an interface structure with barrier function and immune microenvironment characteristics; the interlayer membrane 12 can be made of a porous polymer membrane or hydrogel membrane with a specific pore size and permeability to reconstruct the invasion process of cancer cells across the endothelial barrier; the standard module channel unit chip layer H11 serves as a distal organ bionic module to capture migrated cancer cells and simulate their colonization and diffusion behavior in the distal microenvironment. This embodiment can spatially simulate the continuous process of release from the primary tumor lesion, invasion of the vascular barrier, transmembrane migration, and distal colonization.

[0059] While the embodiments of the present invention have been described in detail above with reference to the accompanying drawings, the present invention is not limited to the embodiments described above. Various modifications may be made within the scope of knowledge possessed by a person skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features within the embodiments may be combined with one another unless there is a conflict.

Claims

1. A modular intelligent breast cancer organ-on-a-chip system, characterized in that: The system includes: a standard module channel unit chip layer and a standard chip channel interface component, which can be freely combined. Different standard module channel unit chip layers or a standard module channel unit chip layer and its mirror-symmetrical standard module channel unit chip layer can be assembled in series or in parallel through the standard chip channel interface component to form the modular intelligent breast cancer organ chip system. The standard module channel unit chip layer is used to realize functions such as cell culture, fluid transport, and material exchange, and includes a microchannel inlet connection, a microchannel chamber, and a microchannel outlet connection; the microchannel inlet connection and the microchannel outlet connection are respectively connected to the two ends of the microchannel chamber; The standard chip channel interface component is used to achieve fluid connection and rapid assembly between different standard module channel unit chip layers, including a transition microchannel and protruding microchannel socket metal connecting pipes located on both sides of the transition microchannel and perpendicular to the two ends of the plane where the transition microchannel is located.

2. A modular intelligent breast cancer organ chip system according to claim 1, characterized in that: The microchannel socket metal connecting tube is respectively inserted into the microchannel inlet connection and the microchannel outlet connection by interference fit; thereby achieving standardized docking and fluid penetration between different standard module channel unit chip layers, and is used to achieve interference fit connection with the channel socket metal connecting tube in the standard chip channel interface assembly.

3. The modular intelligent breast cancer organ chip system according to claim 1, characterized in that: When the standard module channel unit chip layer and its mirror-symmetrical standard module channel unit chip layer are closely combined in parallel with equal sides, the center distance between the microchannel inlet connection of the standard module channel unit chip layer and the microchannel outlet connection of the mirror-symmetrical standard module channel unit chip layer is equal to the center distance between the two microchannel socket metal connecting pipes; When two identical standard module channel unit chip layers are relative to each other and bonded up and down, the distance between the centers of the microchannel inlet connections on the same side of the standard module channel unit chip layer is equal to the distance between the centers of the two microchannel socket metal connecting tubes, and the distance between the centers of the microchannel outlet connections on the same side is equal to the distance between the centers of the two microchannel socket metal connecting tubes.

4. The modular intelligent breast cancer organ chip system according to claim 1, characterized in that: The front and rear ends of the microchannel chamber are respectively connected to the microchannel inlet connection and the microchannel outlet connection through transition channels; the microchannel inlet connection and the microchannel outlet connection are standardized circular reserved structures; the cross section of the transition microchannel is a regular rectangle.

5. The modular intelligent breast cancer organ chip system according to claim 1, characterized in that: A microsensor array is embedded in the standard module channel unit chip layer to realize the intelligent response capability of the modular intelligent breast cancer organ chip system; the microsensor array is one or more of a micropressure sensor, a flow rate sensor, a pH sensor, an oxygen concentration sensor, an electrical impedance sensor, or a membrane displacement sensor; the microsensor array is arranged inside or on the surface of the microchannel chamber, connected to the standard signal interface led out from the edge of the standard module channel unit chip layer, and connected to the external control system; through real-time data acquisition of the microsensor array, the system can sense the local fluid environment, cell metabolic state, or membrane deformation within the standard module channel unit chip layer, and realize automatic adjustment of flow rate, pressure, and chemical factor concentration parameters through linkage with a micropump, a pressure drive device, or an environmental control module.

6. The modular intelligent breast cancer organ chip system according to claim 4, characterized in that: The standard module channel unit chip layer has a length of 6 cm, a width of 3 cm, and a thickness of 0.4 cm; the central microchannel chamber has a rectangular cross-sectional structure with a width of 2 mm, a length of 2 cm, and a height range of 50 to 200 μm; the radius of the standardized circular reserved structures at the microchannel inlet connection and the microchannel outlet connection are both 500 μm; the transition channel width at the front and rear ends of the microchannel chamber is 500 μm.

7. The modular intelligent breast cancer organ chip system according to claim 4, characterized in that: The standard chip channel interface component is 2.2 cm long, 0.5 cm wide, and 0.4 cm thick; the cross-section of the transition microchannel is a regular rectangle with a side length of 500 μm; the microchannel socket metal connecting tubes are all metal conduits with an inner diameter of 1000 μm, and the center distance between the two microchannel socket metal connecting tubes is 1.5 cm.

8. The modular intelligent breast cancer organ chip system according to any one of claims 1 to 7, characterized in that: This includes a combination of the following: Method 1: A fluidic direct-connection docking series model based on a combination of at least two different standard modular channel unit chip layers, comprising: standard modular channel unit chip layers connected in series; the microchannel outlet connections and / or microchannel inlet connections of the two standard modular channel unit chip layers are interference-fitted through the microchannel socket metal connecting tubes of the standard chip channel interface assembly; each of the standard modular channel unit chip layers can achieve independent cell culture, perfusion control, and functional loading, and can be flexibly configured into different functional modules such as a primary tumor lesion module, a vascular module, and a lymphatic module according to research needs; Method 2: A linear chemotactic gradient induction model for substance concentration based on a combination of at least two different standard module channel unit chip layers, comprising: an upper standard module channel unit chip layer and a lower standard module channel unit chip layer; the microchannel inlet connection of the upper layer and the microchannel inlet connection of the lower layer respectively serve as injection ports for two solutions of different concentrations, and the injected fluids are brought into parallel flow contact at the front end through a parallel microchannel chamber of equal width and height; because the fluids in the microchannel chambers are in a low Reynolds number laminar flow state, the two solutions are gradually mixed by molecular diffusion at the interface of the microchannel chambers, forming a stable linear concentration gradient in the axial direction of the channel; Method 3: A dynamic deformation pressure chamber model based on a combination of at least two different standard module channel unit chip layers, comprising: adding an intermediate flexible film layer on the basis of Method 2, wherein the intermediate flexible film layer is arranged between the upper standard module channel unit chip layer and the lower standard module channel unit chip layer, and the upper standard module channel unit chip layer and the lower standard module channel unit chip layer are sequentially bonded after plasma treatment to form a three-layer closed structure, wherein the microchannel chamber of the upper standard module channel unit chip layer and the microchannel chamber of the lower standard module channel unit chip layer are completely isolated after relative bonding; the upper standard module channel unit chip layer The microchannel chamber serves as a pressure chamber and is connected to an external micro air pump via a microfluidic pipeline. It can periodically apply positive or negative pressure according to a preset program, causing the intermediate flexible film layer to bulge or sink into the microchannel chamber of the lower standard module channel unit chip layer below. The intermediate flexible film layer vibrates back and forth to simulate the dynamic effects of vascular flow or lymphatic vessel rhythm on the luminal morphology and fluid shear force. The microchannel chamber of the lower standard module channel unit chip layer is used to culture cells or drain fluids, and the deformation of the intermediate flexible film layer and the fluid dynamics parameters in the channel are monitored in real time via micro displacement sensors and flow rate sensors integrated into the intermediate flexible film layer or the channel wall. Method 4: Freely combine Method 1, Method 2 and Method 3.

9. The method of use according to claims 1 to 8, characterized in that: The specific process is: The tumor cell invasion-culture-transport model based on the combination of at least three different standard module channel unit chip layers includes: a standard module channel unit chip layer for simulating the primary tumor lesion is provided on one side, and a three-layer structure is formed by the lower blood vessel-co-culture module, the middle porous film layer and the upper distal organ bionic module stacked up and down on the other side; the lower blood vessel-co-culture module and the upper distal organ bionic module are both standard module channel unit chip layers; the porous film layer sandwiched in the middle is used to simulate the process of cancer cells crossing the epithelial barrier, which is an important interface connecting the two functional areas; the primary tumor lesion module is connected to the blood vessel-co-culture module through a microchannel socket metal connecting tube, forming a continuous fluid path for the release of front-end tumor cells to the middle blood vessel barrier path; the vascular-co-culture module and the distal organ module are directly bonded through the middle porous film layer to form a vertical migration system with upper and lower dual channels; breast cancer cells are seeded in the primary tumor lesion module to simulate their growth, spread or shedding process in situ; the microchannel chamber of the vascular-co-culture module can be paved with vascular endothelial cells, and immune cells can be introduced when necessary to construct a microenvironment barrier that is closer to the real pathological state; the middle porous film layer can use a porous polymer membrane or hydrogel membrane with a specific pore size and permeability to form a simulated interface with a physiological barrier function, which is used to restore the behavior of cancer cells crossing the endothelial structure during invasion; the distal organ bionic module is used to capture migrated cancer cells and reproduce their colonization and spread environment in distal tissues.

10. The method of use according to claim 9, characterized in that: The vascular-co-culture module can be replaced with a lymphatic network module to simulate the invasion, entry, and migration behavior of breast cancer cells in the lymphatic transport pathway, thereby reconstructing and dynamically monitoring the tumor cell-lymphatic system metastasis mechanism; alternatively, the vascular-co-culture module is connected in series with a standard module channel unit chip layer for simulating the lymphatic pathway through a standard chip channel interface component, thereby sequentially including four different functional areas in one system: the primary tumor lesion module, the vascular module, the lymphatic network module, and the distal organ module, constructing a multi-stage tumor metastasis pathway model covering both the blood and lymphatic pathways, and realizing dynamic monitoring and intervention research of the microenvironment under multi-channel, multi-interface, and multi-factor coupling conditions.

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