Tumor cell culture device and culture method

Abstract

The tumor cell culture device comprises a dish bottom and a dish cover, a culture cavity is formed between the dish bottom and the dish cover, an output pipe is arranged at the dish bottom, and an adjusting assembly used for adjusting the circulation state of the output pipe is arranged on the output pipe; a conveying piece communicated with the container and the culture cavity is arranged between the container and the culture cavity, and the conveying piece is in a closed state when the liquid level of the culture solution in the culture cavity is higher than the communicating position of the conveying piece and the culture cavity; when the liquid level of the culture solution in the culture cavity is lower than the communicating part of the conveying part and the culture cavity, the conveying part is in an open state. When the output pipe gradually discharges the culture solution in the culture cavity, the liquid level height of the culture solution in the culture cavity is reduced, and when the liquid level height of the culture solution is reduced to not exceed the height of the communication position of the conveying part and the culture cavity, the conveying part conveys the culture solution to the culture cavity, and automatic culture cavity liquid supplementing is formed by combining output of the output pipe and input of the conveying part. In the process, manual operation is remarkably reduced, so that the probability of pollution caused by manual uncovering and liquid changing operation is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and in particular to a tumor cell culture device and a culture method. Background Art

[0002] Tumor cells occupy a central position in tissue culture and are an extremely important means of studying the mechanisms of carcinogenesis, anticancer drug testing, and cancer molecular biology. They play an immeasurable role in clarifying and solving cancer.

[0003] Among them, the culture dish is a laboratory dish used for microbial or cell culture, consisting of a dish bottom and a dish cover. Culture medium and tumor cells are placed in the dish bottom, and the dish cover is covered on the dish bottom and then left to culture.

[0004] During the culture process, due to cell metabolism, the operator needs to change the liquid (replace the culture medium) regularly. During the liquid change process, since the lid of the dish needs to be opened to change the liquid, the bottom of the dish will be exposed to the external environment. In order to reduce the impact of the external environment on the cells cultured in the bottom of the dish, the operator needs to perform a series of treatments on the external environment (such as disinfection) before changing the liquid.

[0005] However, changing the liquid in this way requires frequently opening the dish cover, which increases the risk of contamination of the environment at the bottom of the dish. At the same time, frequent operations increase the operator's time consumption and rely on proficiency. Summary of the Invention

[0006] In view of this, the purpose of the present invention is to provide a tumor cell culture device and culture method to solve the problem that the existing culture device needs to frequently open the dish cover when changing the liquid, which increases the risk of contamination of the environment at the bottom of the dish. At the same time, the frequent operation increases the operator's time consumption and depends on the operator's proficiency.

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

[0008] A tumor cell culture device comprises a dish bottom and a dish cover disposed on the dish bottom, forming a culture chamber between the two. The dish bottom is provided with an output tube having one end connected to the culture chamber, and the output tube is provided with a regulating component for regulating the flow state thereof.

[0009] It also includes a container for storing culture fluid, and a conveying member connected to the container and the culture chamber is provided between the container and the culture chamber. When the liquid level of the culture fluid in the culture chamber is higher than the connection point between the conveying member and the culture chamber, the conveying member is in a closed state; when the liquid level of the culture fluid in the culture chamber is lower than the connection point between the conveying member and the culture chamber, the conveying member is in an open state.

[0010] It is further defined that the conveying member includes a first tube body and a second tube body, both ends of the first tube body and the second tube body are connected to the container and the culture chamber respectively, and the distance between the first tube body and the bottom of the dish is greater than the distance between the second tube body and the bottom of the dish.

[0011] It is further defined that an extension tube is provided on a side of the first tube body facing the bottom of the dish, and the extension tube is a frustum-shaped tube body with the tip facing upward.

[0012] It is further defined that the regulating assembly includes a solenoid valve and a liquid level trigger, the upper end of the output tube extends vertically into the culture chamber, the upper end area of ​​the output tube is used to install the liquid level trigger, and the lower end area of ​​the output tube is used to install the solenoid valve, the liquid level trigger is used to monitor the liquid level of the culture medium in the culture chamber and trigger the solenoid valve to enter a flow state when the liquid level reaches a certain threshold;

[0013] A connecting port located between the electromagnetic valve and the liquid level triggering component is opened on the side wall of the output pipe.

[0014] It is further defined that the liquid level trigger includes a movable block and a pressure sensor, the movable block is located in the output pipe and can slide axially along the output pipe, the pressure sensor is located in the output pipe, directly above the movable block, the pressure sensor is located on the sliding track of the movable block, and the pressure sensor is electrically connected to the solenoid valve.

[0015] It is further defined that a limiting ring with an outer diameter smaller than the diameter of the output pipe is provided in the output pipe, the limiting ring is located between the connecting port and the pressure sensor, and the limiting ring is located below the movable block.

[0016] It is further defined that the movable block is a floating body.

[0017] It is further defined that a first magnet is provided on the top surface of the movable block, and a second magnet is provided on the top surface of the dish cover, which is opposite to the first magnet in upper and lower directions, and the polarity of the opposite sides of the first magnet and the second magnet are the same.

[0018] It is further defined that the dish cover is also provided with a rotating rod connected to the second magnet, and one end of the rotating rod away from the second magnet is rotatably connected to the dish cover for adjusting the horizontal position of the second magnet relative to the first magnet.

[0019] A cell culture method comprising the following steps:

[0020] S1: Use two tumor cell culture devices, and set up a glioma cell culture area and a tumor stromal cell culture area in the two tumor cell culture devices respectively;

[0021] S2: Glioma cells are seeded in the glioma cell culture area, and astrocytes and / or oligodendrocytes are seeded in the tumor stromal cell culture area;

[0022] S3: Serum-containing culture medium is injected into the container, and then the culture medium is continuously transported to the culture chamber through the conveying member until the liquid level reaches above the connection point between the conveying member and the culture chamber, forming a static liquid level environment;

[0023] S4: Collect some of the migratory bodies produced by astrocytes and / or oligodendrocytes and place them in the glioma cell culture area;

[0024] S5: After culturing for 48-72 hours, the culture medium is discharged through the output tube, and Matrigel is added to the glioma cell culture area to detect the area of ​​invasion of the glioma cells into the Matrigel.

[0025] The beneficial effects of the present invention are:

[0026] As the output tube gradually discharges the culture fluid in the culture chamber, the liquid level of the culture fluid in the culture chamber decreases accordingly. When the liquid level of the culture fluid drops to a level that does not exceed the height of the connection point between the conveying member and the culture chamber, the conveying member conveys the culture fluid to the culture chamber. The output through the output tube and the input of the conveying member combine to form automatic fluid replenishment of the culture chamber. In this process, manual operation is significantly reduced, thereby reducing the probability of contamination introduced by manual opening of the cover for fluid replacement.

[0027] Other advantages, objects, and features of the present invention will be described in part in the following description and, in part, will be apparent to those skilled in the art upon examination of the following description or may be learned from practice of the present invention. The objects and other advantages of the present invention may be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Schematic diagram of the three-dimensional structure of the present invention (first viewing angle);

[0029] Figure 2 Schematic diagram of the three-dimensional structure of the present invention (second viewing angle);

[0030] Figure 3 A top view of the present invention;

[0031] Figure 4 for Figure 3 Cross-sectional view of AA;

[0032] Figure 5 for Figure 4 The enlarged schematic diagram of point a in the middle;

[0033] Figure 6 Schematic diagram of the internal structure of the output tube of the present invention;

[0034] Figure 7 Schematic diagram of the cell culture method of the present invention.

[0035] In the picture:

[0036] 1. Dish bottom; 101. Output tube; 1011. Connecting port; 2. Dish cover; 3. Container; 4. First tube body; 401. Extension tube; 5. Second tube body; 6. Solenoid valve; 7. Movable block; 8. Pressure sensor; 9. Limiting ring; 10. First magnet; 11. Second magnet; 12. Rotating rod. DETAILED DESCRIPTION

[0037] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Generally, the components of the embodiments of the present invention described and shown in the drawings herein can be arranged and designed in various different configurations.

[0038] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention as claimed, but rather merely represents selected embodiments of the present invention. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without creative effort shall fall within the scope of protection of the present invention.

[0039] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, it does not require further definition or explanation in subsequent drawings.

[0040] In the above description of the present invention, it should be noted that the terms "one side," "the other side," and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, or the orientations or positional relationships in which the inventive product is typically placed when in use. These terms are intended solely to facilitate the description of the present invention and simplify the description, and are not intended to indicate or imply that the devices or components referred to must have, be constructed, or operate in a specific orientation. Therefore, they should not be construed as limitations on the present invention. Furthermore, the terms "first," "second," and the like are used solely for distinction and should not be construed as indicating or implying relative importance.

[0041] Furthermore, the term "identical" and similar terms do not necessarily require that the components be absolutely identical; slight variations are permitted. The term "perpendicular" simply refers to the positional relationship between components being more perpendicular than "parallel," not that the structure must be perfectly vertical; rather, it can be slightly tilted.

[0042] See also Figure 1-7The present invention provides a technical solution: a tumor cell culture device, comprising a dish bottom and a dish cover disposed on the dish bottom, forming a culture chamber between the two, the dish bottom being provided with an output tube having one end connected to the culture chamber, the output tube being provided with a regulating component for regulating its flow state;

[0043] It also includes a container for storing culture fluid, and a conveying member connected to the container and the culture chamber is provided between the container and the culture chamber. When the liquid level of the culture fluid in the culture chamber is higher than the connection point between the conveying member and the culture chamber, the conveying member is in a closed state; when the liquid level of the culture fluid in the culture chamber is lower than the connection point between the conveying member and the culture chamber, the conveying member is in an open state.

[0044] The dish cover is a breathable cover, and a breathable membrane can be provided on the dish cover to enable gas exchange between the culture chamber and the outside through the breathable membrane, thereby providing air for the cells to be cultured.

[0045] In this solution, the conveyor is designed to be linked to the culture fluid level. When the culture fluid exceeds the height of the connection point between the conveyor and the culture chamber, the conveyor stops delivering culture fluid to the culture chamber. Conversely, when the culture fluid does not exceed the height of the connection point between the conveyor and the culture chamber, the conveyor continues to deliver culture fluid to the culture chamber. This mechanism can reduce the risk of excessive addition of culture fluid leading to hypoxia of the cultured cells or accumulation of metabolic waste.

[0046] The output tube cooperates with the regulating member, and the flow state of the output tube can be adjusted by the regulating member (ie, the culture chamber can / cannot discharge the culture medium in the culture chamber through the output tube).

[0047] As the output tube gradually discharges the culture fluid in the culture chamber, the liquid level of the culture fluid in the culture chamber decreases accordingly. When the liquid level of the culture fluid drops to a level that does not exceed the height of the connection point between the conveying member and the culture chamber, the conveying member conveys the culture fluid to the culture chamber. The output through the output tube and the input of the conveying member combine to form automatic fluid replenishment of the culture chamber. In this process, manual operation is significantly reduced, thereby reducing the probability of contamination introduced by manual opening of the cover for fluid replacement.

[0048] Through the combination of conveying parts, adjusting parts and output pipe, it can be used in two ways:

[0049] Usage method 1: Regularly open the output tube through the adjustment part to discharge the culture fluid in the culture chamber, and automatically replenish the fluid in the culture chamber through the conveying part. The output tube can be continuously opened and closed by the adjustment part to promote the accumulation of a certain amount of culture fluid in the culture chamber and then discharge it again, thereby repeatedly achieving a certain cleaning effect;

[0050] Usage method 2: The output tube is continuously opened through the adjustment part, and the culture fluid in the culture chamber is continuously discharged. The culture fluid automatically replenished through the conveying part is also discharged through the output tube. This method forms a dynamic culture environment in the culture chamber. The continuous flow of culture fluid can drive the efficiency of gas exchange in the culture chamber, so that the stimulation to the cells is improved through the flowing culture fluid and increased gas exchange (this premise is based on the cells being cultured for a period of time and the cells adhering to the inner wall of the bottom of the dish).

[0051] In this embodiment, the conveying member includes a first tube body and a second tube body, both ends of which are connected to the container and the culture chamber respectively, and the distance between the first tube body and the bottom of the dish is greater than the distance between the second tube body and the bottom of the dish.

[0052] In this solution, the first tube body and the second tube body are both "L"-shaped tubes, with two horizontal sections connected to the container and two vertical sections connected to the culture chamber. The horizontal height of the end of the first tube body connected to the culture chamber (referred to as port A) is lower than the horizontal height of the end of the second tube body connected to the culture chamber (referred to as port B). In this way, the first tube body serves as a high-level tube for transporting air, and the second tube body serves as a low-level tube for transporting culture fluid. The container is a sealed container and is connected to the outside world only through the first tube body and the second tube body.

[0053] Since the level of port B is lower than that of port A, when the culture liquid level exceeds port A, port B will inevitably be submerged by the culture liquid in the culture chamber. In this state, according to the principle of communicating vessels, the culture liquid in the container cannot be transported to the culture chamber through the second tube.

[0054] As the output tube discharges the old culture fluid in the culture chamber, the liquid level in the culture chamber gradually drops until port A is exposed above the culture fluid surface. The first tube body is connected to the outside air. Under the action of atmospheric pressure, the culture fluid in the container can be normally transported to the culture chamber through the second tube body, thereby forming automatic fluid replenishment.

[0055] Specifically, the first tube (high-level tube) serves as an air channel, while the second tube (low-level tube) serves as a fluid infusion channel. This system utilizes the principle of a communicating vessel to automatically replenish culture fluid without the need for external power. Furthermore, the high-level tube ensures that fluid infusion only occurs when the liquid level drops below the low-level tube port, preventing backflow of culture fluid and contamination of the container.

[0056] In this embodiment, an extension tube is provided on a side of the first tube body facing the bottom of the dish, and the extension tube is a frustum-shaped tube body with the tip facing upward.

[0057] In this solution, the first tube body has an opening expanded by the outwardly expanding extension tube to reduce blockage. Furthermore, if bubbles are generated by collision during the transportation of the culture fluid, the bubbles can be decomposed by the non-uniform force on the conical surface.

[0058] In this embodiment, the regulating assembly includes a solenoid valve and a liquid level trigger. The upper end of the output tube extends vertically into the culture chamber. The upper end area of ​​the output tube is used to install the liquid level trigger, and the lower end area is used to install the solenoid valve. The liquid level trigger is used to monitor the liquid level of the culture medium in the culture chamber and trigger the solenoid valve to enter a flow state when the liquid level reaches a certain threshold.

[0059] A connecting port located between the electromagnetic valve and the liquid level triggering component is opened on the side wall of the output pipe.

[0060] In this solution, the upper end of the output tube extends into the culture chamber, creating a height difference with the inner bottom surface of the dish. This height difference also creates a triggering condition for the liquid-level trigger. The output tube and the culture chamber are connected via a connector, allowing culture fluid in the culture chamber to enter the output tube through the connector and be discharged through a solenoid valve. When the solenoid valve is closed, culture fluid enters the output tube and accumulates, causing the culture fluid level in the output tube to rise. This rise, when it reaches a certain height, triggers the liquid-level trigger to activate the solenoid valve to open.

[0061] In this embodiment, the liquid level trigger includes a movable block and a pressure sensor. The movable block is located in the output pipe and can slide axially along the output pipe. The pressure sensor is located in the output pipe and directly above the movable block. The pressure sensor is located on the sliding track of the movable block. The pressure sensor is electrically connected to the solenoid valve.

[0062] In this solution, the movable block has a diameter similar to that of the output tube and is located within it. The two slide together, enabling the movable block to slide axially along the tube. The inner wall of the tube limits the movable block, enhancing its stability during movement. A pressure sensor is located along the movable block's sliding path, and mechanical triggering occurs when the movable block slides to a specific position.

[0063] In this embodiment, a limiting ring with an outer diameter smaller than the diameter of the output tube is provided in the output tube. The limiting ring is located between the connecting port and the pressure sensor, and the limiting ring is located below the movable block.

[0064] In this solution, because the diameter of the movable block is similar to that of the output tube, a stop ring with a smaller outer diameter is installed inside the output tube to limit the movable block's sliding range. The stop ring is located above the connection port, so the stop ring restricts the movable block's sliding range to just above the connection port. This restrictive action prevents the movable block from sliding into the connection port and potentially blocking the output tube.

[0065] In this embodiment, the movable block is a floating body (eg a lightweight shell with a hollow interior).

[0066] In this solution, the output tube is connected to the culture chamber via a connection port. Therefore, when the culture fluid level in the culture chamber rises, the culture fluid level in the output tube rises synchronously. When it reaches a certain height, the traditional limit ring contacts the bottom surface of the movable block. Because the movable block has the characteristics of a float, as the culture fluid level continues to rise, it pushes the movable block upward, thereby causing the movable block to slide upward in the output tube. When it slides to a certain height, it contacts the pressure sensor, which triggers the solenoid valve to open and discharge the culture fluid. As the culture fluid is discharged, the culture fluid level in the output tube drops, and the movable block, no longer supported by the culture fluid, also drops synchronously, causing the movable block to slide downward in the output tube. As the culture fluid level in the culture chamber drops, the first tube body is exposed, causing the container to be affected by atmospheric pressure, and the second tube body once again transports culture fluid into the culture chamber.

[0067] A relay unit connected to the pressure sensor and solenoid valve can be added. When the pressure sensor is squeezed by the movable block, it sends an activation signal to the relay. The relay receives the activation signal and, after a delay, transmits a start signal to the solenoid valve, which then opens to discharge the culture fluid. This discharge process involves a delay, allowing the new culture fluid delivered through the second tube to remain within the culture chamber for a period of time. During this delay, the new culture fluid can submerge itself within the first tube, rather than being discharged directly, facilitating cleaning of the chamber.

[0068] In this embodiment, a first magnet is provided on the top surface of the movable block, and a second magnet is provided on the top surface of the dish cover, which is opposite to the first magnet in vertical direction. The polarity of the opposite sides of the first magnet and the second magnet are the same.

[0069] In this solution, since the polarity of the first magnet and the second magnet on one side relative to each other is the same, they can generate a downward thrust on the movable block through the repulsion between the same poles. Under the action of the thrust, the movable block overcomes the supporting force brought by the rising culture fluid, preventing the movable block from contacting the pressure sensor, thereby ensuring that the solenoid valve will not be opened and the culture fluid in the culture chamber will not be discharged.

[0070] In this embodiment, a rotating rod connected to the second magnet is further provided on the dish cover. One end of the rotating rod away from the second magnet is rotatably connected to the dish cover for adjusting the horizontal position of the second magnet relative to the first magnet.

[0071] In this solution, a rotating lever is rotatably connected to the lid. The operator manually moves the lever to move the second magnet toward or away from the first magnet. Once the second magnet moves away from the first magnet, the first magnet loses its thrust. Driven by buoyancy and the culture fluid level, the movable block slides upward again and contacts the pressure sensor, triggering the solenoid valve to open and drain the culture fluid.

[0072] A cell culture method comprising the following steps:

[0073] S1: Use two tumor cell culture devices, and set up a glioma cell culture area and a tumor stromal cell culture area in the two tumor cell culture devices respectively;

[0074] S2: Glioma cells are inoculated into the glioma cell culture area.

[0075] Astrocytes and / or oligodendrocytes are seeded into the tumor stromal cell culture area, and astrocytes and / or oligodendrocytes are fluorescently labeled with TSPAN3 / 14 / 15 to facilitate migratory body tracking;

[0076] S3: DMEM / F12 culture medium containing 10% FBS is injected into the container. The culture medium is then continuously delivered to the culture chamber via the delivery device until the liquid level reaches above the connection point between the delivery device and the culture chamber, forming a static liquid surface environment. The interstitial cell region is pre-treated with a cell migration inducer (e.g., EGF) for 48 hours to promote the formation of migrasomes (migrasome formation mechanism).

[0077] S4: Collect conditioned medium from the interstitial cell culture area, separate migrasomes by differential centrifugation plus density gradient centrifugation, and place the collected migrasomes in the glioma cell culture area;

[0078] S5: After 48-72 hours of culture, drain the culture medium through the outlet tube and add Matrigel to the lower glioma cell area with a uniform thickness (1 mm). Continue culturing for 24 hours and use fluorescence microscopy (if the cells have been labeled) or Calcein-AM live cell staining to calculate the depth (μm) of glioma cell invasion into the Matrigel and measure the percentage of invasion area.

[0079] This method first proves that astrocytes / oligodendrocytes drive glioma invasion through migrasomes (non-traditional EVs) through the astrocyte / oligodendrocyte-migrasome-glioma invasion mechanism, providing new evidence for the "tumor-microenvironment interaction" theory; and locks in TSPAN3 / 14 / 15 as migrasome-specific markers, providing a precise target for blocking migrasome generation; elucidating the core reason for the unclear boundaries of gliomas (occult invasion mediated by migrasomes), breaking through the limitations of traditional imaging boundary determination.

[0080] That is, by standardizing the isolation and invasion quantification of migratoria, a reliable experimental model is provided for subsequent therapeutic strategies targeting migratoria (such as blocking TSPAN proteins).

[0081] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A tumor cell culture device comprising a dish bottom (1) and a dish cover (2) disposed on the dish bottom (1), forming a culture chamber therebetween, characterized in that: The bottom of the dish (1) is provided with an output tube (101) having one end in communication with the culture chamber, and the output tube (101) is provided with a regulating component for regulating its flow state; The apparatus further comprises a container (3) storing a culture fluid, wherein a conveying member communicating with the container (3) and the culture chamber is provided between the container (3) and the culture chamber, wherein the conveying member is in a closed state when the level of the culture fluid in the culture chamber is higher than the connection point between the conveying member and the culture chamber; and the conveying member is in an open state when the level of the culture fluid in the culture chamber is lower than the connection point between the conveying member and the culture chamber.

2. The tumor cell culture device according to claim 1, characterized in that: The conveying member comprises a first tube body (4) and a second tube body (5), the two ends of the first tube body (4) and the second tube body (5) are respectively connected to the container (3) and the culture chamber, and the distance between the first tube body (4) and the bottom of the dish (1) is greater than the distance between the second tube body (5) and the bottom of the dish (1).

3. The tumor cell culture device according to claim 2, characterized in that: An extension tube (401) is provided on the side of the first tube body (4) facing the dish bottom (1), and the extension tube (401) is a frustum-shaped tube body with the tip facing upward.

4. The tumor cell culture device according to claim 1, wherein: The regulating assembly comprises a solenoid valve (6) and a liquid level trigger. The upper end of the output tube (101) extends vertically into the culture chamber. The upper end region of the output tube (101) is used to install the liquid level trigger, and the lower end region is used to install the solenoid valve (6). The liquid level trigger is used to monitor the liquid level of the culture medium in the culture chamber and trigger the solenoid valve (6) to enter a flow state when the liquid level reaches a certain threshold. A connecting port (1011) located between the electromagnetic valve (6) and the liquid level triggering component is provided on the side wall of the output pipe (101).

5. The tumor cell culture device according to claim 4, characterized in that: The liquid level triggering component comprises a movable block (7) and a pressure sensor (8); the movable block (7) is located in the output pipe (101) and can slide axially along the output pipe (101); the pressure sensor (8) is located in the output pipe (101) and directly above the movable block (7); the pressure sensor (8) is located on the sliding track of the movable block (7); and the pressure sensor (8) is electrically connected to the solenoid valve (6).

6. The tumor cell culture device according to claim 4 or 5, characterized in that: A limiting ring (9) having an outer diameter smaller than the diameter of the output tube (101) is provided in the output tube (101), the limiting ring (9) being located between the connecting port (1011) and the pressure sensor (8), and the limiting ring (9) being located below the movable block (7).

7. The tumor cell culture device according to claim 6, characterized in that: The movable block (7) is a floating body.

8. The tumor cell culture device according to claim 6, characterized in that: The top surface of the movable block (7) is provided with a first magnet (10), and the top surface of the dish cover (2) is provided with a second magnet (11) which is opposite to the first magnet (10) in the upper and lower directions. The polarities of the opposite sides of the first magnet (10) and the second magnet (11) are the same.

9. The tumor cell culture device according to claim 8, characterized in that: The dish cover (2) is also provided with a rotating rod (12) connected to the second magnet (11); one end of the rotating rod (12) away from the second magnet (11) is rotatably connected to the dish cover (2) for adjusting the horizontal position of the second magnet (11) relative to the first magnet (10).

10. A cell culture method using the tumor cell culture device according to any one of claims 1 to 9, comprising the following steps: S1: Use two tumor cell culture devices, and set up a glioma cell culture area and a tumor stromal cell culture area in the two tumor cell culture devices respectively; S2: Glioma cells are seeded in the glioma cell culture area, and astrocytes and / or oligodendrocytes are seeded in the tumor stromal cell culture area; S3: injecting serum-containing culture medium into the container (3), and then continuously delivering the culture medium to the culture chamber through the conveying member until the liquid level reaches above the connection point between the conveying member and the culture chamber, forming a static liquid level environment; S4: Collect some of the migratory bodies produced by astrocytes and / or oligodendrocytes and place them in the glioma cell culture area; S5: After culturing for 48-72 hours, the culture medium is discharged through the output tube (101), and Matrigel is added to the glioma cell culture area to detect the invasion area of ​​the glioma cells into the Matrigel.

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