Cancer cell migration and angiogenesis-based anti-cancer drug screening chip and method

Abstract

The invention belongs to the technical field of microfluidic organ chips, and particularly relates to an anti-cancer drug screening chip and method based on cancer cell migration and angiogenesis, the organ chip comprises at least one monitoring culture module, and the monitoring culture module comprises a vascular endothelial cell culture layer, a PET porous membrane layer and a tumor cell culture layer which are fixedly connected in sequence; the vascular endothelial cell culture layer comprises a vascular endothelial cell culture chamber, and two sides of the vascular endothelial cell culture chamber are respectively communicated with a liquid inlet I and a liquid outlet I; the tumor cell culture layer comprises a tumor cell culture chamber, one side of the tumor cell culture chamber is communicated with a liquid inlet II, the tumor cell culture chamber and the vascular endothelial cell culture chamber are correspondingly arranged and communicated, and the PET porous membrane layer is arranged between the vascular endothelial cell culture chamber and the tumor cell culture chamber; one side of the migration chamber is communicated with the other side of the tumor cell culture chamber, and the other side of the migration chamber is communicated with a liquid outlet II.

Description

Technical Field

[0001] The present invention belongs to the technical field of microfluidic organ chips, and in particular relates to an anticancer drug screening chip and method based on cancer cell migration and angiogenesis. Background Art

[0002] Cancer cell migration and tumor angiogenesis are two crucial biological processes in the development and metastasis of cancer. These two processes not only play a key role in the growth and spread of tumors, but are also important targets for anti-cancer therapeutic strategies. Traditional research methods, including cell experiments under two-dimensional and three-dimensional conditions and in vivo experiments based on experimental animals, still have significant limitations.

[0003] Specifically, two-dimensional cell culture technology cannot reproduce the complex in vivo tumor microenvironment, especially the cytokine and signal transduction processes related to tumor development. Although three-dimensional cell culture, such as the common tumor spheroid culture, is closer to physiological conditions, it still cannot fully simulate the shear stress, signal transduction between multiple cell types, and cell-to-cell interactions under physiological conditions. Although animal experimental models provide a microenvironment that is closer to the physiological state, due to species differences, the results cannot be perfectly extrapolated to the human body.

[0004] In response to the above problems, microfluidic organ chip technology came into being, which can simulate and study the tumor microenvironment at the micrometer scale. Some advanced studies have more accurately restored the tumor-vascular interface through three-dimensional cell culture in microfluidic organ chips, thereby better simulating the tumor microenvironment in the human body. However, current technology has not been able to achieve simultaneous monitoring of the two key functions of tumor angiogenesis and tumor cell migration on a single microfluidic chip, which limits the ability to simultaneously evaluate changes in tumor angiogenesis and cell migration functions under the influence of drugs in related drug screening studies. Therefore, there is an urgent need for anticancer drug screening chips and methods based on cancer cell migration and angiogenesis to solve this problem. Summary of the invention

[0005] The purpose of the present invention is to provide a chip and method for screening anticancer drugs based on cancer cell migration and angiogenesis to solve the above problems.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] A microfluidic organ chip for anticancer drug screening comprises at least one monitoring and culture module, wherein the monitoring and culture module comprises:

[0008] A vascular endothelial cell culture layer, a PET porous membrane layer and a tumor cell culture layer fixed in sequence;

[0009] The vascular endothelial cell culture layer comprises:

[0010] A vascular endothelial cell culture chamber, wherein two sides of the vascular endothelial cell culture chamber are respectively connected with a liquid inlet 1 and a liquid outlet 1;

[0011] The tumor cell culture layer comprises:

[0012] A tumor cell culture chamber, one side of which is connected to a second liquid inlet, the tumor cell culture chamber is arranged correspondingly to and connected to the vascular endothelial cell culture chamber, and the PET porous membrane layer is arranged between the vascular endothelial cell culture chamber and the tumor cell culture chamber;

[0013] One side of the migration chamber is connected to the other side of the tumor cell culture chamber, and the other side of the migration chamber is connected to a second liquid outlet.

[0014] Optionally, a fluid channel one is provided between the vascular endothelial cell culture chamber and the liquid inlet one, one end of the fluid channel one is connected to the vascular endothelial cell culture chamber, and the other end of the fluid channel one is connected to the liquid inlet one.

[0015] Optionally, a fluid channel 2 is provided between the vascular endothelial cell culture chamber and the liquid outlet 1, one end of the fluid channel 2 is connected to the vascular endothelial cell culture chamber, and the other end of the fluid channel 2 is connected to the liquid outlet 1.

[0016] Optionally, a fluid channel three is provided between the tumor cell culture chamber and the liquid inlet two, one end of the fluid channel three is connected to the tumor cell culture chamber, and the other end of the fluid channel three is connected to the liquid inlet two.

[0017] Optionally, a fluid channel four is provided between the tumor cell culture chamber and the migration chamber, one end of the fluid channel four is connected to the tumor cell culture chamber, and the other end of the fluid channel four is connected to the migration chamber.

[0018] Optionally, a fluid channel four is provided between the tumor cell culture chamber and the migration chamber, one end of the fluid channel four is connected to the tumor cell culture chamber, and the other end of the fluid channel four is connected to the migration chamber.

[0019] Optionally, a piston for controlling the opening and closing of the fluid channel 4 is provided in the middle of the fluid channel 4, and the piston is slidably connected with a hole, and the hole penetrates the vascular endothelial cell culture layer.

[0020] Optionally, a fluid channel five is provided between the migration chamber and the second liquid outlet, one end of the fluid channel five is connected to the migration chamber, and the other end of the fluid channel five is connected to the second liquid outlet.

[0021] Optionally, the liquid inlet end of the second liquid inlet is arranged to penetrate the vascular endothelial cell culture layer.

[0022] Optionally, the liquid outlet end of the second liquid outlet is arranged to penetrate the vascular endothelial cell culture layer.

[0023] A method for screening anticancer drugs, using the above anticancer drug to screen a microfluidic organ chip, comprises the following steps:

[0024] disinfect;

[0025] Inoculating different types of cells in the vascular endothelial cell culture layer and the tumor cell culture layer;

[0026] Cell culture;

[0027] Pharmacological interventions;

[0028] Observe and analyze the results of drug intervention.

[0029] Compared with the prior art, the present invention has the following advantages and technical effects:

[0030] When in use, the microfluidic organ chip is first disinfected, and different types of cells are inoculated in the vascular endothelial cell culture layer and the tumor cell culture layer, and the cells are cultured and intervened with drugs. Since the vascular endothelial cells and tumor cells are located in different chambers, the two types of cells are separated by a PET porous membrane layer with a specific pore size, and the PET porous membrane layer allows the two types of cells to communicate information under simulated physiological conditions, providing a highly bionic in vivo environment simulation platform for drug screening. In traditional models, tumor cells must pass through a barrier with a larger pore size and invade the vascular endothelial cell layer to determine their metastatic ability. This chip sets a special migration chamber in the same plane as the tumor cell culture layer, which can directly observe the migration behavior of tumor cells without crossing the membrane structure. This design not only simplifies the evaluation of tumor cell migration ability, but also allows tumor cells to migrate without direct contact with the vascular endothelial layer, avoiding the direct impact of tumor cells on the vascular endothelial cell layer. In addition, the chip can also realize the simultaneous monitoring of tumor cell migration and angiogenesis, providing independent observation windows for research. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative labor:

[0032] Figure 1 It is an exploded view of the structure of the present invention;

[0033] Figure 2 A bottom view of the vascular endothelial cell culture layer of the present invention;

[0034] Figure 3 A top view of the tumor cell culture layer of the present invention;

[0035] Figure 4 It is a top perspective view of the structure of the present invention;

[0036] Among them, 1. vascular endothelial cell culture layer; 2. PET porous membrane layer; 3. tumor cell culture layer; 4. liquid inlet one; 5. vascular endothelial cell culture chamber; 6. liquid outlet one; 7. fluid channel one; 8. fluid channel two; 9. punching; 10. liquid inlet two; 11. tumor cell culture chamber; 12. migration chamber; 13. liquid outlet two; 14. fluid channel three; 15. fluid channel four; 16. fluid channel five. DETAILED DESCRIPTION

[0037] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in 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. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0039] Reference Figures 1 to 4 The present invention discloses a microfluidic organ chip for anticancer drug screening, comprising at least one monitoring and culture module, wherein the monitoring and culture module comprises:

[0040] A vascular endothelial cell culture layer 1, a PET porous membrane layer 2 and a tumor cell culture layer 3 fixed in sequence;

[0041] The vascular endothelial cell culture layer 1 comprises:

[0042] The vascular endothelial cell culture chamber 5 has a liquid inlet 4 and a liquid outlet 6 connected to the two sides of the vascular endothelial cell culture chamber 5;

[0043] Tumor cell culture layer 3 includes:

[0044] A tumor cell culture chamber 11, one side of which is connected to a liquid inlet 2 10, the tumor cell culture chamber 11 is arranged correspondingly and connected to the vascular endothelial cell culture chamber 5, and the PET porous membrane layer 2 is arranged between the vascular endothelial cell culture chamber 5 and the tumor cell culture chamber 11;

[0045] One side of the migration chamber 12 is connected to the other side of the tumor cell culture chamber 11 , and the other side of the migration chamber 12 is connected to a second liquid outlet 13 .

[0046] When in use, the microfluidic organ chip is first disinfected, and different types of cells are inoculated in the vascular endothelial cell culture layer 1 and the tumor cell culture layer 3, respectively, and the cells are cultured and intervened with drugs. Since the vascular endothelial cells and tumor cells are located in different chambers, the two types of cells are isolated by a PET porous membrane layer 2 with a specific pore size, and the PET porous membrane layer 2 allows the two types of cells to communicate information under simulated physiological conditions, providing a highly bionic in vivo environment simulation platform for drug screening. In traditional models, tumor cells must pass through a barrier with a larger pore size and invade the vascular endothelial cell layer to determine their metastatic ability. This chip sets a special migration chamber 12 in the same plane of the tumor cell culture layer, which can directly observe the migration behavior of tumor cells without crossing the membrane structure. Such a design not only simplifies the evaluation of tumor cell migration ability, but also allows tumor cells to migrate without direct contact with the vascular endothelial layer, avoiding the direct impact of tumor cells on the vascular endothelial cell layer. In addition, the chip can also realize the simultaneous monitoring of tumor cell migration and angiogenesis, providing independent observation windows for research.

[0047] As an optional embodiment, a fluid channel 7 is provided between the vascular endothelial cell culture chamber 5 and the liquid inlet 4, one end of the fluid channel 7 is connected to the vascular endothelial cell culture chamber 5, and the other end of the fluid channel 7 is connected to the liquid inlet 4.

[0048] As an optional embodiment, a fluid channel 2 8 is provided between the vascular endothelial cell culture chamber 5 and the liquid outlet 1 6 , one end of the fluid channel 2 8 is connected to the vascular endothelial cell culture chamber 5 , and the other end of the fluid channel 2 8 is connected to the liquid outlet 1 6 .

[0049] As an optional embodiment, a fluid channel three 14 is provided between the tumor cell culture chamber 11 and the liquid inlet two 10 , one end of the fluid channel three 14 is connected to the tumor cell culture chamber 11 , and the other end of the fluid channel three 14 is connected to the liquid inlet two 10 .

[0050] As an optional embodiment, a fluid channel 415 is provided between the tumor cell culture chamber 11 and the migration chamber 12, one end of the fluid channel 415 is connected to the tumor cell culture chamber 11, and the other end of the fluid channel 415 is connected to the migration chamber 12;

[0051] A piston for controlling the opening and closing of the fluid channel 4 15 is provided in the middle of the fluid channel 4 15 , and the piston is slidably connected with a perforation 9 , which is arranged through the vascular endothelial cell culture layer 1 .

[0052] As an optional embodiment, a fluid channel five 16 is provided between the migration chamber 12 and the second liquid outlet 13 , one end of the fluid channel five 16 is connected to the migration chamber 12 , and the other end of the fluid channel five 16 is connected to the second liquid outlet 13 .

[0053] As an optional implementation, the liquid inlet end of the second liquid inlet 10 is arranged to penetrate the vascular endothelial cell culture layer 1 .

[0054] As an optional implementation, the liquid outlet end of the second liquid outlet 13 is arranged to penetrate the vascular endothelial cell culture layer 1 .

[0055] This embodiment provides a microfluidic organ chip for screening anticancer drugs based on cancer cell migration and angiogenesis. The microfluidic organ chip provided in this embodiment has four identical monitoring and culture modules, which are arranged in mirror symmetry with a central axis to ensure the uniformity of the design and the consistency of the experiment.

[0056] The monitoring culture module consists of three main structures: a vascular endothelial cell culture layer 1, a PET porous membrane layer 2, and a tumor cell culture layer 3.

[0057] The vascular endothelial cell culture layer 1 is located at the top layer, the PET porous membrane layer 2 is located in the middle layer, and the tumor cell culture layer 3 is located at the bottom layer. The three are combined through precise alignment and bonding technology to form an overall three-layer composite structure. The final size of the four monitoring culture modules is 75mm×25mm.

[0058] In the vascular endothelial cell culture layer 1, liquid culture medium can be injected through the liquid inlet 1 4, flow through the vascular endothelial cell culture chamber 5, and finally flow out through the liquid outlet 1 6. Each part is connected by the corresponding fluid channel 1 7 and fluid channel 2 8.

[0059] In the design of the vascular endothelial cell culture layer 1, the vascular endothelial cell culture chamber 5 has a high degree of design flexibility. It can be used as a top-closed chamber, where liquid is injected and discharged only through the fluid channel; or a small hole can be opened at the top of the chamber to directly inject liquid, which is suitable for three-dimensional culture media such as hydrogels; in addition, the chamber can also be designed to be completely open, which is convenient for organoid culture.

[0060] The vascular endothelial cell culture chamber 5 can be designed as a closed chamber similar to the tumor cell culture chamber 11, that is, the chamber is only connected to the fluid channel 1 7 and the fluid channel 2 8, and the top of the chamber is not exposed to the air;

[0061] Opening a small hole means making a small hole in the center of a closed chamber. It can be simply understood as adding an inlet / outlet to the top of the chamber, which plays the same role. This design is only for the convenience of experimental operation. Therefore, it can be flexibly selected according to the specific experimental plan.

[0062] Regardless of the structure, there is only a PET porous membrane between the vascular endothelial cell culture chamber 5 and the tumor cell culture chamber 11, allowing liquid exchange.

[0063] This design provides the possibility for a variety of experimental environments and enhances the application range of the chip. In this example, the vascular endothelial cell culture chamber 5 is a circular chamber with a diameter of 8 mm, which is close to the size of a standard 96-well plate, ensuring that there is enough sample volume for subsequent analysis. The diameter of the chamber can be flexibly modified according to experimental requirements. The length, width and depth of fluid channel 1 7 and fluid channel 2 8 can be flexibly adjusted according to experimental requirements and processing conditions to provide suitable liquid flow conditions.

[0064] In the tumor cell culture layer 3, the liquid culture medium can be injected through the second liquid inlet 10, and flow through the tumor cell culture chamber 11 and the migration chamber 12 in sequence, and finally flow out through the second liquid outlet 13. The size of the tumor cell culture chamber 11 is consistent with the diameter of the vascular endothelial cell culture chamber 5 in the vascular endothelial cell culture layer 1, both of which are 8mm, and are aligned between the upper and lower layers to achieve effective cell signal communication in the multilayer structure. The specific size of the migration chamber 12 is 10mm×2mm, and a smooth rectangular edge is used in the design. This design helps to reduce turbulence and dead corners generated during the liquid flow process, making the flow of the fluid in the migration chamber 12 more uniform and stable.

[0065] The tumor cell culture chamber 11 and the migration chamber 12 are connected through a fluid channel 4 15. A switch is provided in the middle of the fluid channel 4 15. The switch can be controlled by: a hole 9 is provided at a corresponding position in the vascular endothelial cell culture layer 1, and a hard cylindrical piston of a suitable size is embedded at the position. When the piston is fully inserted into the fluid channel 4 15, the fluid channel 4 15 is closed; when the plug is pulled out, the fluid channel 4 15 is opened.

[0066] The vascular endothelial cell culture layer 1 and the tumor cell culture layer 3 in the chip are both provided with independent liquid inlets and outlets, so that suitable culture media can be selected according to different cell types, providing the best culture environment for the upper and lower layers. In addition, the operator can carry out different types of cell culture according to experimental needs.

[0067] The material and specification of the PET porous membrane layer 2 are adjustable. In this example, a PET porous membrane with a pore size of 4 microns and a thickness of 12 microns is selected to meet the requirements of cancer cell migration and angiogenesis experiments for material exchange.

[0068] As an additional implementation, the method for manufacturing the microfluidic organ chip comprises the following steps:

[0069] Step 1: Design and carve the mold. Use computer-aided design (CAD) software to design the structural layout of the endothelial cell culture chamber 5 and the tumor cell culture chamber 11, and carve the corresponding chambers and fluid channels through soft lithography technology. You can also choose to output to micromachining equipment for mold processing. The purpose of this step is to ensure that each component of the microfluidic organ chip can be accurately matched.

[0070] Step 2: Prepare the PDMS layer. Mix the polydimethylsiloxane (PDMS) prepolymer and the curing agent in a certain proportion, and remove the bubbles in the mixture. Pour the mixed PDMS into the mold obtained in step 1 and solidify it to form a PDMS layer with a specific depth of fluid channels and chambers.

[0071] Furthermore, the ratio of polydimethylsiloxane (PDMS) prepolymer to curing agent is 10:1.

[0072] Step 3: Prepare PET porous membrane. According to the experimental requirements, select a polyethylene terephthalate (PET) membrane with appropriate pore size and thickness for microporous processing to form pores that allow cytokines and small molecules to pass through. The purpose of this step is to achieve material exchange between vascular endothelial cells and tumor cells while maintaining physical isolation between the two cells.

[0073] Step 4: Assemble the chip. The assembly of the microfluidic organ chip is completed by aligning and bonding the PDMS layer and the porous PET membrane in a standardized process. This step requires ensuring the sealing between the layers to prevent the non-specific passage of culture medium or cells between different culture chambers.

[0074] A method for screening anticancer drugs, using the above anticancer drug to screen a microfluidic organ chip, comprises the following steps:

[0075] disinfect;

[0076] Different types of cells are inoculated on the vascular endothelial cell culture layer 1 and the tumor cell culture layer 3;

[0077] Cell culture;

[0078] Pharmacological interventions;

[0079] Observe and analyze the results of drug intervention.

[0080] The specific implementation steps of this example for anticancer drug screening for cancer cell migration and angiogenesis are as follows:

[0081] Step 1: Chip pretreatment: Before the experiment, the chip was placed in a clean bench and exposed to ultraviolet light for 30 minutes, and each chamber and channel was rinsed with phosphate buffer solution (PBS) under sterile conditions.

[0082] Step 2: Cell inoculation. Based on the structure of the chip, different types of cells can be cultured in the endothelial cell culture layer 1 and the tumor cell culture layer 3. In order to promote cell adhesion and growth, 0.01-0.1 mg / mL rat tail collagen or 0.01-0.1% poly-lysine can be used in the chamber for modification in advance. The specific coating time should be flexibly adjusted according to the experimental requirements, ranging from about 10 minutes to several hours.

[0083] For cell inoculation of endothelial cell culture layer 1, first place the matrix gel in ice and melt at 4°C. Use a pre-cooled pipette or pipette tip to mix until uniform, then add DMEM culture medium to dilute the matrix gel to the required concentration. Add the diluted matrix gel and then incubate at 37°C for 1 hour to solidify the matrix gel. Digest, resuspend and count human umbilical vein endothelial cells (HUVECs) with a confluence of 70-80%, and adjust the cell density to 1×10 5 / mL, inject the hydrogel solution into the vascular endothelial cell culture chamber on ice for 5.

[0084] For the cell inoculation of the tumor cell culture layer 3, the fluid channel 4 15 was first closed, and the closing method was as described above. Then, human cervical squamous cell carcinoma cells with a confluence of 70-80% were digested, resuspended and counted, and the cell density was adjusted to 1×10 5 / mL, and inject the cell suspension through the inlet port = 10. After inoculating the cells, the chip was placed in an incubator at 37°C with 5% carbon dioxide to allow the SiHa cells to adhere to the wall.

[0085] Step 3: Cell culture and drug intervention. During cell culture, use a syringe pump to inject liquid culture medium into the liquid inlet 14 and the liquid inlet 2 10, and connect the corresponding pipelines to the liquid outlet 16 and the liquid outlet 2 13 for collecting the effluent. For the semi-open and open vascular endothelial cell culture chambers 5, a sealing film can be used to seal them during the culture process to prevent the volatilization of the liquid. The chip is then placed in an incubator at 37°C and containing 5% carbon dioxide for culture. When performing drug intervention, the drug is diluted in the liquid culture medium to prepare a working solution of a specific concentration, and intervention is performed in the same manner as cell culture.

[0086] Step 4: Observe the effect of drug intervention on tumor angiogenesis. After 24 hours of drug intervention, remove the existing culture medium, stain with DMEM culture medium containing live cell dye calcein AM, take photos under a microscope, and observe the formation of vascular network.

[0087] Step 5: Observe the effect of drug intervention on the migration function of cancer cells. 24 hours after drug intervention, observe the number of cells in the migration chamber and the distance of cell migration under the bright field of a microscope to detect the effect of drug intervention on the migration function of cancer cells.

[0088] Step 6: Collect the effluent for analysis of angiogenesis indicators. During step 3, collect the effluent under cell culture and drug intervention for quantitative detection of cytokines. A commercial antibody chip can be selected to analyze the level of angiogenesis factors in the effluent to determine changes in angiogenesis-related functions.

[0089] Step 7: Cell digestion and Western blot experiment. After the culture is completed, PBS buffer is injected into the inlet 14 and the inlet 210 for washing 2-3 times. After washing, the buffer is removed and trypsin is added for digestion. After the digestion is terminated with the corresponding complete culture medium, the cell suspension is collected for subsequent Western blot experiments.

[0090] Compared with the traditional technology, the present invention has the following advantages:

[0091] (1) Multifunctional cell culture and interface simulation: The chip has different chambers for culturing endothelial cells and tumor cells. The two types of cells are separated by a porous membrane with a specific pore size. The membrane structure design allows the two cells to communicate information under simulated physiological conditions, providing a highly bionic in vivo environment simulation platform for drug screening.

[0092] (2) Independent migration chamber design: The chip includes a migration chamber, which is connected to the tumor cell culture chamber through a fluid channel with a controllable switch. It is different from the traditional experimental design that evaluates the tumor metastasis ability through the transmembrane invasion behavior of tumor cells. In the traditional model, tumor cells must pass through a barrier with a larger pore size and invade the vascular endothelial cell layer to determine their metastasis ability. This chip sets a special migration chamber in the same plane as the tumor cell culture layer to directly observe the migration behavior of tumor cells without crossing the membrane structure. This design not only simplifies the evaluation of tumor cell migration ability, but also allows tumor cells to migrate without direct contact with the vascular endothelial layer, avoiding the direct impact of tumor cells on the vascular endothelial cell layer. In addition, the chip can also realize the simultaneous monitoring of tumor cell migration and angiogenesis, providing independent observation windows for research.

[0093] (3) Optimized operating interface and multi-dimensional culture: The chamber design of the present invention is flexible. A closed or top-open chamber structure can be used according to experimental requirements to simplify actual operations. This design facilitates the use of scaffold materials such as hydrogels in the chip to culture vascular endothelial cells and observe phenomena such as the sprouting of new blood vessels in real time. It also facilitates the implantation and transfer of organoids, enhancing the application potential of the chip in drug testing. The dual-channel design of the chip is widely applicable to cell culture of various types and different stages.

[0094] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside" and "outside" etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0095] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. An anticancer drug screening chip based on cancer cell migration and angiogenesis, characterized in that: It includes at least one monitoring and cultivation module, and the monitoring and cultivation module includes: A vascular endothelial cell culture layer (1), a PET porous membrane layer (2) and a tumor cell culture layer (3) fixed in sequence; The vascular endothelial cell culture layer (1) comprises: A vascular endothelial cell culture chamber (5), wherein two sides of the vascular endothelial cell culture chamber (5) are respectively connected with a liquid inlet (4) and a liquid outlet (6); The tumor cell culture layer (3) comprises: A tumor cell culture chamber (11), one side of the tumor cell culture chamber (11) being connected to a second liquid inlet (10), the tumor cell culture chamber (11) being arranged correspondingly to and connected to the vascular endothelial cell culture chamber (5), and the PET porous membrane layer (2) being arranged between the vascular endothelial cell culture chamber (5) and the tumor cell culture chamber (11); One side of the migration chamber (12) is connected to the other side of the tumor cell culture chamber (11), and the other side of the migration chamber (12) is connected to a second liquid outlet (13).

2. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 1, characterized in that: A fluid channel (7) is provided between the vascular endothelial cell culture chamber (5) and the liquid inlet (4), one end of the fluid channel (7) is connected to the vascular endothelial cell culture chamber (5), and the other end of the fluid channel (7) is connected to the liquid inlet (4).

3. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 1, characterized in that: A fluid channel 2 (8) is provided between the vascular endothelial cell culture chamber (5) and the liquid outlet 1 (6), one end of the fluid channel 2 (8) is connected to the vascular endothelial cell culture chamber (5), and the other end of the fluid channel 2 (8) is connected to the liquid outlet 1 (6).

4. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 1, characterized in that: A fluid channel three (14) is provided between the tumor cell culture chamber (11) and the second liquid inlet (10), one end of the fluid channel three (14) is connected to the tumor cell culture chamber (11), and the other end of the fluid channel three (14) is connected to the second liquid inlet (10).

5. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 1, characterized in that: A fluid channel four (15) is provided between the tumor cell culture chamber (11) and the migration chamber (12), one end of the fluid channel four (15) is connected to the tumor cell culture chamber (11), and the other end of the fluid channel four (15) is connected to the migration chamber (12).

6. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 5, characterized in that: A piston for controlling the opening and closing of the fluid channel four (15) is provided in the middle of the fluid channel four (15), and the piston is slidably connected to a perforation (9), and the perforation (9) is arranged through the vascular endothelial cell culture layer (1).

7. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 1, characterized in that: A fluid channel five (16) is provided between the migration chamber (12) and the second liquid outlet (13), one end of the fluid channel five (16) is connected to the migration chamber (12), and the other end of the fluid channel five (16) is connected to the second liquid outlet (13).

8. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 1, characterized in that: The liquid inlet end of the second liquid inlet (10) is arranged to penetrate the vascular endothelial cell culture layer (1).

9. The anticancer drug screening chip based on cancer cell migration and angiogenesis according to claim 8, characterized in that: The liquid outlet end of the second liquid outlet (13) is arranged to penetrate the vascular endothelial cell culture layer (1).

10. A method for screening anticancer drugs, using the anticancer drug screening chip based on cancer cell migration and angiogenesis according to any one of claims 1 to 9, characterized in that: The steps include: disinfect; Inoculating different types of cells in the vascular endothelial cell culture layer (1) and the tumor cell culture layer (3); Cell culture; Pharmacological interventions; Observe and analyze the results of drug intervention.

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