Extracorporeal circulation type organ 3D culture device based on tumor microenvironment

Abstract

The invention discloses an extracorporeal circulation organoid 3D culture device based on a tumor microenvironment. The extracorporeal circulation organoid 3D culture device comprises a circulation driving module, a sample adding module and a 3D cell tissue culture module. The circulation driving module is a key for constructing extracorporeal circulation and drives a culture solution to flow in the device so as to achieve the purpose of simulating continuous flow of blood in a human body; the sample adding module is used for helping a culture solution in the device to metabolize and ensuring that components of the culture solution are normal all the time so as to achieve the effect of simulating blood components in a human body; the 3D cell tissue culture module is used for providing a culture environment with a 3D framework for the growth of tumor cells so as to ensure that the cells are cultured in the culture environment with the 3D framework. The circulation driving module, the sample adding module and the 3D cell tissue culture module are communicated through pipelines, so that a complete in-vivo blood circulation system is formed, better 3D in-vitro cell culture can be carried out, and the influence of blood circulation on tumor cell culture is effectively simulated.

Description

Technical Field

[0001] The patent of this invention relates to the technical field of in vitro cell culture, and specifically to an in vitro circulation organoid 3D culture device based on tumor microenvironment. Background Art

[0002] The tumor cell microenvironment is an extremely complex and sophisticated system that includes not only the tumor cells themselves, but also a variety of closely related cell types and their various microenvironments, which are further subdivided into immune microenvironment, microbial microenvironment, microcirculatory microenvironment, and metabolic microenvironment. These components together shape a dynamic and balanced ecosystem that plays a decisive role in the growth, invasion, and metastasis of tumors.

[0003] In the basic research of tumor organoids, technicians in this field gradually realized that it is far from enough to focus only on tumor cells themselves. In fact, other types of cells closely related to tumors are equally numerous and crucial. These cells include organ-intrinsic cells, which constitute the basic framework of tumor organoids; mesenchymal cells, such as vascular endothelial cells and fibroblasts, which play a key role in tumor angiogenesis and matrix remodeling; and immune cells, such as T cells, B cells, and macrophages, which affect tumor progression through complex immune responses. These different types of cells jointly construct the microenvironment of tumor organoids through a complex network of interactions, thereby determining the biological behavior of the tumor.

[0004] However, current research methods are powerless when faced with such a complex system. Due to the limitations of experimental equipment and technology, most existing research is limited to 2D in vitro cell culture, where tumor cells are cultured on two-dimensional surfaces such as flat culture dishes. The interaction between cells is relatively limited, and due to the limitations of culture conditions, only the relationship between two cells at most can be studied, or it can be simplified to the analysis of the interaction between one cell and another. This simplified research model obviously cannot truly reflect the complexity of the tumor organoid microenvironment, so the conclusions drawn are often one-sided and may mislead our understanding of tumor biological mechanisms.

[0005] Another issue worth noting is that the current in vitro cell culture system has significant deficiencies in simulating the in vivo blood circulation system. Blood circulation is an important part of the tumor organoid microenvironment. Blood provides cells with necessary nutrients and oxygen, and is also responsible for the excretion of metabolic waste. In real tumor tissue, blood circulation has a great impact on it, ensuring that tumor cells obtain sufficient nutrients and oxygen to support their rapid growth. However, in in vitro cell culture, due to the lack of an effective blood circulation simulation system, cells are often in a state of malnutrition and accumulation of metabolic waste, which seriously affects the growth state of cells and the accuracy of experimental results.

[0006] Therefore, there is a need to provide an in vitro circulation organoid 3D culture device based on the tumor microenvironment that can perform 3D in vitro cell culture and simulate the effect of blood circulation on tumor cells. Summary of the invention

[0007] In order to solve the problems raised in the above background technology, the present invention provides a 3D culture device of in vitro circulation organoids based on tumor microenvironment, which solves the problem that it is difficult to carry out 3D in vitro cell culture in the current technical field of in vitro cell culture and it is impossible to simulate the effect of blood circulation on tumor cells.

[0008] To achieve the above-mentioned purpose, the present invention provides the following technical solution: an in vitro circulation organoid 3D culture device based on tumor microenvironment, comprising:

[0009] A circulation drive module, used for providing circulation power for the culture liquid in the culture device;

[0010] A sample loading module, including a sample loading opening, for adding a cell suspension and / or an intervention reagent into the culture device, and adjusting the composition of the culture solution to meet the needs of the physiological processes of growth, differentiation and migration of cells in the culture device;

[0011] 3D cell tissue culture modules, including spheroid culture dishes, are used to create a 3D culture environment and provide a place for the physiological processes of cell growth, differentiation and migration within the culture device;

[0012] The circulation drive module, the sample loading module and the 3D cell tissue culture module are connected by a pipeline, and each includes a liquid inlet and a liquid outlet, wherein the liquid outlet of the sample loading module is connected to the liquid inlet of the circulation drive module, the liquid outlet of the circulation drive module is connected to the liquid inlet of the 3D cell tissue culture module, and the liquid outlet of the 3D cell tissue culture module is connected to the liquid inlet of the sample loading module.

[0013] Based on the above-mentioned in vitro circulation organoid 3D culture device based on tumor microenvironment, in one feasible design, the circulation drive module includes a circulation machine and a drive unit;

[0014] The driving unit is arranged outside the pipeline, and the driving end of the driving unit is fixedly connected to the rotating shaft of the circulator, wherein the circulator is arranged in the pipeline and is used to rotate and drive the culture fluid in the pipeline to circulate.

[0015] Based on the above-mentioned extracorporeal circulation organoid 3D culture device based on tumor microenvironment, in a feasible design, the sample loading opening is arranged on the surface of the pipeline, and an opening cover is movably connected to the sample loading opening, and the opening cover covers the sample loading opening when closed.

[0016] Based on the above-mentioned in vitro circulation organoid 3D culture device based on tumor microenvironment, in an achievable design, the liquid inlet and liquid outlet of the 3D cell tissue culture module are arranged at corresponding positions of the spherical culture dish, at least one spherical culture dish is provided, each spherical culture dish is connected by a pipeline, and an isolation membrane is provided at the position of the liquid inlet and liquid outlet of any spherical culture dish;

[0017] The isolation membrane covers the liquid inlet and the liquid outlet of the spherical culture dish, and is used to construct a non-contact co-culture space for the tumor cells in the spherical culture dish.

[0018] Based on the above-mentioned extracorporeal circulation organoid 3D culture device based on tumor microenvironment, in a feasible design, the isolation membrane adopts a polycarbonate porous membrane, and the pore size of the polycarbonate porous membrane is set to 0um-3um.

[0019] Based on the above-mentioned in vitro circulation organoid 3D culture device based on tumor microenvironment, in a feasible design, the 3D cell tissue culture module also includes a culture dish slot and a timed rotation buckle;

[0020] The spherical culture dish is snapped into the culture dish slot via the timing rotation snap-in, the timing rotation snap-in is arranged on the outer surface of the spherical culture dish, and the timing rotation snap-in is used for timing control of the rotation of the spherical culture dish.

[0021] Based on the above-mentioned extracorporeal circulation organoid 3D culture device based on tumor microenvironment, in an achievable design, it is characterized in that it also includes a pipeline sealing valve, which is used to ensure the circulation stability of the culture solution in the culture device and the sterility of the culture environment;

[0022] The pipeline sealing valves include at least three, which are respectively arranged in the pipeline between the liquid outlet of the sample adding module and the liquid inlet of the circulation driving module, in the pipeline between the liquid outlet of the circulation driving module and the liquid inlet of the 3D cell tissue culture module, and in the pipeline between the liquid outlet of the 3D cell tissue culture module and the liquid inlet of the sample adding module.

[0023] Based on the above-mentioned extracorporeal circulation organoid 3D culture device based on tumor microenvironment, in a feasible design, it is characterized in that it also includes a main control module, which is used to integrate the control and coordinate the work of the circulation drive module, the sampling module and the 3D cell tissue culture module, wherein the control end of the main control module is electrically connected to the controlled ends of the circulation drive module, the sampling module and the 3D cell tissue culture module, respectively.

[0024] Based on the above-mentioned extracorporeal circulation organoid 3D culture device based on tumor microenvironment, in a feasible design, it is characterized in that it also includes a display interaction module, which is electrically connected to the main control module, and is used to monitor the culture environment parameters in the culture device in real time and display them visually, and to issue instructions to the main control module during human-computer interaction, so that the main control module outputs a control signal to the circulation drive module, the sample loading module and the 3D cell tissue culture module to adjust the culture environment parameters in the culture device, wherein the culture environment parameters include the circulation flow rate of the culture fluid.

[0025] Based on the above-mentioned in vitro circulation organoid 3D culture device based on tumor microenvironment, in an achievable design, the display interaction module includes a sampling sensor unit, a display unit and an interaction unit, wherein the sampling sensor unit is arranged in the pipeline;

[0026] The feedback signal output end of the sampling sensor unit is electrically connected to the feedback signal input end of the main control module, the controlled end of the display unit is electrically connected to the control end of the main control module, and is used to visually display the culture environment parameters, and the command signal output end of the interactive unit is electrically connected to the command signal input end of the main control module.

[0027] Beneficial effects:

[0028] The in vitro circulation organoid 3D culture device based on tumor microenvironment disclosed by the present invention includes a circulation drive module, a sample loading module and a 3D cell tissue culture module. The circulation drive module is the key to constructing the in vitro circulation, which drives the culture fluid to flow in the device to achieve the purpose of simulating the continuous flow of blood in the human body; the sample loading module is to help the culture fluid in the device to metabolize, and always ensure that the composition of the culture fluid is normal, so as to achieve the effect of simulating the blood components in the human body; the 3D cell tissue culture module is used to provide a 3D structured culture environment for the growth of tumor cells, so as to ensure that the cells are cultured in a 3D structured culture environment. The circulation drive module, the sample loading module and the 3D cell tissue culture module are connected by pipelines, which forms a complete in vivo blood circulation system, which can perform better 3D in vitro cell culture, and effectively simulate the influence of blood circulation on tumor cell culture, helping scientific researchers to better explore the interaction between tumor cells and the growth progress of tumor cells in the actual body. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a functional structural block diagram of the in vitro circulation organoid 3D culture device based on tumor microenvironment in Example 1 of the present invention;

[0030] Figure 2 This is a functional structural block diagram of the in vitro circulation organoid 3D culture device based on tumor microenvironment in Example 2 of the present invention;

[0031] Figure 3 This is a schematic diagram of the overall structure of the in vitro circulation organoid 3D culture device based on the tumor microenvironment in Example 2 of the present invention;

[0032] Figure 4 This is a schematic diagram of the structure of the circulation driving module of the in vitro circulation organoid 3D culture device based on the tumor microenvironment in Example 2 of the present invention;

[0033] Figure 5 This is a schematic diagram of a sample loading module of the in vitro circulation organoid 3D culture device based on tumor microenvironment in Example 2 of the present invention;

[0034] Figure 6 This is a schematic diagram of the structure of a 3D cell tissue culture module of an in vitro circulation organoid 3D culture device based on a tumor microenvironment in Example 2 of the present invention;

[0035] Figure 7 This is a structural schematic diagram of the display interaction module of the in vitro circulation organoid 3D culture device based on tumor microenvironment in Example 2 of the present invention.

[0036] In the figure, 1. circulation drive module; 2. sample loading module; 3. 3D cell tissue culture module; 4. display interaction module; 5. sample loading opening; 6. spherical culture dish; 7. circulation machine; 8. drive unit; 9. culture dish slot; 10. pipeline; 11. opening cover. DETAILED DESCRIPTION

[0037] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the present invention will be briefly introduced below in combination with the drawings and the description of the embodiments or the prior art. Obviously, the following description of the structure of the drawings is only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention.

[0038] It should be understood that although the terms first, second, etc. may be used herein to describe various units, these units should not be limited by these terms. These terms are only used to distinguish one unit from another unit. For example, a first unit can be referred to as a second unit, and similarly, a second unit can be referred to as a first unit without departing from the scope of the exemplary embodiments of the present invention.

[0039] It should be understood that the term "and / or" that may appear in this article is merely a description of the association relationship between associated objects, indicating that three relationships may exist. For example, A and / or B can represent three situations: A exists alone, B exists alone, and A and B exist at the same time. The term " / and" that may appear in this article describes another type of association object relationship, indicating that two relationships may exist. For example, A / and B can represent two situations: A exists alone, and A and B exist alone. In addition, the character " / " that may appear in this article generally indicates that the previous and next associated objects are in an "or" relationship.

[0040] Embodiment 1:

[0041] like Figure 1As shown, the present embodiment provides an extracorporeal circulation organoid 3D culture device based on tumor microenvironment, which adopts a circulation drive module 1 as the construction basis of the circulation in the whole device, and uses the circulation drive module 1 to drive the culture fluid in the device to circulate, so that the culture fluid keeps flowing to simulate the real blood circulation environment in the body, so that researchers can explore the influence of blood circulation on tumor cell growth; the spherical culture dish 6 in the 3D cell tissue culture module 3 is used to replace the flat culture dish in the 2D culture to build a 3D culture environment, which will more realistically simulate the growth environment in the body and better present the real environment of cell growth, so that researchers can more accurately explore the cell growth situation; the sample addition module 2 is used to help form a real in vivo blood circulation environment, ensure that each cycle can take away the metabolites in the culture fluid, and add new culture fluid intervention reagents, so that the culture fluid in the next cycle can still keep the composition unchanged, and realistically simulate the blood environment in the body to meet the needs of the physiological process of cell growth, differentiation and migration in the culture device.

[0042] The present embodiment provides an in vitro circulation organoid 3D culture device based on tumor microenvironment, comprising a circulation drive module 1, a sample loading module 2 and a 3D cell tissue culture module 3. Among them, the circulation drive module 1, the sample loading module 2 and the 3D cell tissue culture module 3 are connected through a pipeline 10 to form a complete circulation structure; the circulation drive module 1, the sample loading module 2 and the 3D cell tissue culture module 3 all include a liquid inlet and a liquid outlet, and each module is connected in turn through the liquid outlet and the liquid inlet, specifically: the liquid outlet of the sample loading module 2 is connected to the liquid inlet of the circulation drive module 1, the liquid outlet of the circulation drive module 1 is connected to the liquid inlet of the 3D cell tissue culture module 3, and the liquid outlet of the 3D cell tissue culture module 3 is connected to the liquid inlet of the sample loading module 2.

[0043] Embodiment 2:

[0044] like Figure 2-7 As shown, the present embodiment provides an in vitro circulation organoid 3D culture device based on tumor microenvironment. As a preferred embodiment, the circulation drive module 1 includes a circulation machine 7 and a drive unit; the drive unit is arranged outside the pipeline 10, and the driving end of the drive unit is fixedly connected to the rotating shaft of the circulation machine 7, wherein the circulation machine 7 is arranged in the pipeline 10, and is used to rotate and drive the culture fluid in the pipeline 10 to circulate. The driving unit can use a servo motor with a controllable preferred speed, and its driving shaft is the driving end. When it is started, the driving shaft drives the rotating shaft of the circulation machine 7 to rotate, and the circulation machine 7 can push the culture fluid to flow in a fixed direction to form a cycle; and the circulation machine 7 can choose a rotating mechanism with blades, etc., which is not limited in the present embodiment, and it only needs to achieve the purpose of rotating and driving the liquid flow in the pipeline 10.

[0045] In the present embodiment, a 3D culture device of in vitro circulation organoids based on tumor microenvironment is provided. As a preferred embodiment, the sample loading opening 5 is arranged on the surface of the pipeline 10, and an opening cover 11 is movably connected to the sample loading opening 5. When the opening cover 11 is closed, it covers the sample loading opening 5. This is to ensure the airtightness of the whole device during normal circulation. The opening cover 11 needs to be opened only when loading or discharging metabolites in the original culture solution, so as to help better realize the construction of a real blood environment.

[0046] The present embodiment provides an extracorporeal circulation organoid 3D culture device based on tumor microenvironment. As a preferred embodiment, the liquid inlet and liquid outlet of the 3D cell tissue culture module 3 are arranged at the corresponding positions of the spherical culture dish 6. The spherical culture dish 6 is provided with at least one, and each spherical culture dish 6 is connected by a pipe 10, and an isolation membrane is arranged at the position of the liquid inlet and liquid outlet of any spherical culture dish 6; the isolation membrane covers the liquid inlet and liquid outlet of the spherical culture dish 6, and is used to construct a non-contact co-culture space for tumor cells in the spherical culture dish 6. The isolation membrane adopts a polycarbonate porous membrane, and the pore size of the polycarbonate porous membrane is set to 0um-3um. The setting of this isolation membrane forms a non-contact co-culture environment between each spherical culture dish 6, so that researchers can explore the mechanism of interaction between multiple tumor cell microenvironments, and provide technical support for single-cell sequencing of tumor cell microenvironments and tumor metastasis research. The isolation membrane is a polycarbonate porous membrane, and the pore size of the polycarbonate porous membrane is set to 0um-3um to ensure that cells cannot penetrate the isolation membrane between each spherical culture dish 6, but the culture solution can penetrate to form an overall culture area. The pore size is set to a range of 0um-3um. During specific implementation, specific selection can be made according to needs to simulate interactions between different types of cells or drug intervention.

[0047] The present embodiment provides an extracorporeal circulation organoid 3D culture device based on tumor microenvironment. As a preferred embodiment, the 3D cell tissue culture module 3 also includes a culture dish slot 9 and a timed rotation buckle; the spherical culture dish 6 is connected to the culture dish slot 9 through the timed rotation buckle, and the timed rotation buckle is set on the outer surface of the spherical culture dish 6. The timed rotation buckle is used to control the rotation of the spherical culture dish 6. Between the double-sided hole molds of the spherical culture dish 6, this timed rotation buckle is set to drive the spherical culture dish 6 to rotate in a clockwise direction at a fixed time. In specific use, it can be set to rotate once every 10 minutes, and rotate once in a clockwise direction by rotation. This is to maintain the uniform distribution of cells in the spherical culture dish 6, and if there are multiple spherical culture dishes 6, it can help to achieve the circulation and communication of the culture fluid between each spherical culture dish 6.

[0048] The present embodiment provides an extracorporeal circulation organoid 3D culture device based on tumor microenvironment, as a preferred embodiment, further comprising a pipeline 10 sealing valve, which is used to ensure the circulation stability of the culture solution in the culture device and the sterility of the culture environment; the pipeline 10 sealing valve includes at least three, and is respectively arranged in the pipeline 10 between the liquid outlet of the sample loading module 2 and the liquid inlet of the circulation driving module 1, in the pipeline 10 between the liquid outlet of the circulation driving module 1 and the liquid inlet of the 3D cell tissue culture module 3, and in the pipeline 10 between the liquid outlet of the 3D cell tissue culture module 3 and the liquid inlet of the sample loading module 2. The modules are connected by a sealing interface to ensure the stability and sterility of the culture environment.

[0049] The present embodiment provides an in vitro circulation organoid 3D culture device based on tumor microenvironment, as a preferred embodiment, further comprising a main control module, which is used for integrated control and coordination of the circulation drive module 1, the sample loading module 2 and the 3D cell tissue culture module 3, wherein the control end of the main control module is electrically connected to the controlled end of the circulation drive module 1, the sample loading module 2 and the 3D cell tissue culture module 3. The main control module can make it possible to complete the electrical control between the modules, and form a coordinated whole to better cooperate with each other, for example: when controlling the sample loading module 2 to load or remove the working process of metabolites, the circulation drive module is controlled to stop working at the same time.

[0050] It also includes a display interaction module 4, which is electrically connected to the main control module, and is used to monitor the culture environment parameters in the culture device in real time and display them visually, and to issue instructions to the main control module during human-computer interaction, so that the main control module outputs control signals to the circulation drive module 1, the sample loading module 2 and the 3D cell tissue culture module 3 to adjust the culture environment parameters in the culture device, wherein the culture environment parameters include the circulation flow rate of the culture solution. The display interaction module 4 provides a window for human-computer interaction, and can control the operation of each module through instructions during human-computer interaction, for example: issuing an opening instruction for the sample loading opening 5 or a closing instruction for the sample loading opening 5 through the display interaction module 4, issuing a start instruction for the circulation drive module 1 or a pause instruction for the circulation drive module 1 through the display interaction module 4, issuing a rotation instruction for the spherical culture dish 6 through the display interaction module 4, etc.

[0051] The display interaction module 4 includes a sampling sensor unit, a display unit and an interaction unit, wherein the sampling sensor unit is arranged in the pipeline 10; the feedback signal output end of the sampling sensor unit is electrically connected to the feedback signal input end of the main control module, the controlled end of the display unit is electrically connected to the control end of the main control module, and is used to visualize the culture environment parameters, and the command signal output end of the interaction unit is electrically connected to the command signal input end of the main control module. The display unit can intuitively display the culture environment parameters to the researchers, the most important of which is the circulation flow rate of the culture solution, in addition to which basic parameters such as the number of culture solution circulations can also be included, in order to facilitate the researchers to observe and control the working status of each module through the main control module accordingly.

[0052] Finally, it should be noted that the above are only preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. An in vitro circulation organoid 3D culture device based on tumor microenvironment, characterized in that: include: A circulation drive module, used for providing circulation power for the culture liquid in the culture device; A sample loading module, including a sample loading opening, for adding a culture solution intervention reagent into the culture device, and for discharging metabolites in the culture solution to adjust the composition of the culture solution; 3D cell tissue culture modules, including spheroid culture dishes, are used to create a 3D culture environment and provide a place for the physiological processes of cell growth, differentiation and migration within the culture device; The circulation drive module, the sample loading module and the 3D cell tissue culture module are connected by a pipeline, and each includes a liquid inlet and a liquid outlet, wherein the liquid outlet of the sample loading module is connected to the liquid inlet of the circulation drive module, the liquid outlet of the circulation drive module is connected to the liquid inlet of the 3D cell tissue culture module, and the liquid outlet of the 3D cell tissue culture module is connected to the liquid inlet of the sample loading module.

2. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 1, characterized in that: The circulation drive module includes a circulation machine and a drive unit; The driving unit is arranged outside the pipeline, and the driving end of the driving unit is fixedly connected to the rotating shaft of the circulator, wherein the circulator is arranged in the pipeline and is used to rotate and drive the culture fluid in the pipeline to circulate.

3. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 1, characterized in that: The sample adding opening is arranged on the surface of the pipeline, and an opening cover is movably connected to the sample adding opening. When the opening cover is closed, it covers the sample adding opening.

4. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 1, characterized in that: The liquid inlet and the liquid outlet of the 3D cell tissue culture module are arranged at corresponding positions of the spherical culture dish, at least one spherical culture dish is provided, each spherical culture dish is connected by a pipeline, and an isolation membrane is provided at the position of the liquid inlet and the liquid outlet of any spherical culture dish; The isolation membrane covers the liquid inlet and the liquid outlet of the spherical culture dish, and is used to construct a non-contact co-culture space for the tumor cells in the spherical culture dish.

5. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 4, characterized in that: The isolation membrane is a polycarbonate porous membrane, and the pore size of the polycarbonate porous membrane is set to 0um-3um.

6. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 1, characterized in that: The 3D cell tissue culture module also includes a culture dish slot and a timed rotation buckle; The spherical culture dish is snapped into the culture dish slot via the timing rotation snap-in, the timing rotation snap-in is arranged on the outer surface of the spherical culture dish, and the timing rotation snap-in is used for timing control of the rotation of the spherical culture dish.

7. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 1, characterized in that: It also includes a pipeline sealing valve, which is used to ensure the circulation stability of the culture solution in the culture device and the sterility of the culture environment; The pipeline sealing valves include at least three, which are respectively arranged in the pipeline between the liquid outlet of the sample adding module and the liquid inlet of the circulation driving module, in the pipeline between the liquid outlet of the circulation driving module and the liquid inlet of the 3D cell tissue culture module, and in the pipeline between the liquid outlet of the 3D cell tissue culture module and the liquid inlet of the sample adding module.

8. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 1, characterized in that: It also includes a main control module, which is used to integrate control and coordinate the work of the circulation drive module, the sample loading module and the 3D cell tissue culture module, wherein the control end of the main control module is electrically connected to the controlled ends of the circulation drive module, the sample loading module and the 3D cell tissue culture module respectively.

9. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 8, characterized in that: It also includes a display interaction module, which is electrically connected to the main control module and is used to monitor the culture environment parameters in the culture device in real time and display them visually, and to issue instructions to the main control module during human-computer interaction, so that the main control module outputs control signals to the circulation drive module, the sample loading module and the 3D cell tissue culture module to adjust the culture environment parameters in the culture device, wherein the culture environment parameters include the circulation flow rate of the culture solution.

10. The in vitro circulation organoid 3D culture device based on tumor microenvironment according to claim 8, characterized in that: The display interaction module comprises a sampling sensor unit, a display unit and an interaction unit, wherein the sampling sensor unit is arranged in the pipeline; The feedback signal output end of the sampling sensor unit is electrically connected to the feedback signal input end of the main control module, the controlled end of the display unit is electrically connected to the control end of the main control module, and is used to visually display the culture environment parameters, and the command signal output end of the interactive unit is electrically connected to the command signal input end of the main control module.

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