Tumor tissue model establishment method based on organ-like chip

Abstract

The invention discloses a tumor tissue model establishment method based on an organ-like chip, and relates to the technical field of tumor tissue models. The method comprises the following steps: S1, selecting a proper cell line or tissue source: firstly, selecting a proper tumor cell line or a tumor tissue sample obtained from the body of a patient; s2, designing an organ-like chip: designing a structure of the organ-like chip based on existing tumor biological knowledge and an engineering principle; and S3, preparing a biological material on the chip: culturing and implanting the selected tumor cells or tissues into corresponding positions of the organ-like chip. According to the invention, a proper tumor cell line or a tumor tissue sample obtained from the body of a patient is selected, and the structure of an organ-like chip is designed based on existing tumor biological knowledge and an engineering principle, so that the stability and repeatability of an experiment are ensured, and the experiment efficiency is improved. This contributes to more authentically reflecting individual differences and patient-specific biological characteristics.

Description

Technical Field

[0001] The present invention belongs to the technical field of tumor tissue models, and in particular relates to a method for establishing a tumor tissue model based on an organoid chip. Background Art

[0002] Patient-derived organoids (PDO) are a new tool for studying tumors. In addition to retaining the original biological characteristics of the tumor, they can also be stably propagated. Therefore, they have unique advantages in the development and screening of tumor drugs. In addition, organoids were once named one of the top ten technologies of the year by Science, becoming a powerful tool in the fields of new drug development, precision treatment, and regenerative medicine. In addition to active drug screening and cytotoxicity testing, patient-derived tumor organoids can also provide reliable in vitro tumor models for precision medicine. More and more evidence confirms that the phenotype and genotype between the original tumor tissue and the generated tumor organoids are consistent. At the same time, clinical studies have shown that tumor organoids have a high success rate in predicting individual patients' clinical responses to colorectal cancer and gastroesophageal cancer treatment. However, drug testing based on tumor organoids still takes weeks or even months to provide results to patients.

[0003] Organoids-on-Chips are an organ physiological microsystem built on a chip. With microfluidic chips as the core, they can simulate and construct in vitro tissue and organ microenvironments containing a variety of living cells, functional tissue interfaces, biofluids, mechanical force stimulation and other complex factors, reflecting the main structural and functional characteristics of human tissues and organs. Organoids-on-Chips can be widely used in drug screening, disease simulation, personalized medicine, toxicity testing and other fields, and have a very broad market prospect.

[0004] People are eager to obtain a method for establishing a tumor tissue model based on organoid chips, which can not only solve the problem of long drug testing cycle of traditional tumor organoids, but also simulate the real microenvironment of the tumor in vitro and improve the accuracy of drug sensitivity experiments. Summary of the invention

[0005] The purpose of the present invention is to provide a method for establishing a tumor tissue model based on an organoid chip, which uses a microfluidic chip as a platform, designs the structure of the organoid chip, and constructs a simulated tumor microenvironment under near-physiological conditions in vitro to ensure the stability and repeatability of the experiment, which helps to more realistically reflect individual differences and patient-specific biological characteristics and solve existing problems.

[0006] In order to solve the above technical problems, the present invention is achieved through the following technical solutions:

[0007] The present invention provides a method for establishing a tumor tissue model based on an organoid chip, comprising the following steps:

[0008] S1: Select appropriate cell lines or tissue sources: First, you need to select appropriate tumor cell lines or tumor tissue samples obtained from patients;

[0009] S2: Design organoid chips: Based on existing knowledge of tumor biology and engineering principles, design tumor organoid chips that mimic the tumor microenvironment;

[0010] The structure of the organoid chip is as follows:

[0011] It comprises a lower fluid channel chip, a middle porous membrane layer, an upper organoid culture chip and a cover of the upper chip, wherein the lower fluid channel chip comprises an anti-collapse column structure, and the lower fluid channel chip and the upper organoid culture chip are connected via a porous membrane; tumor organoids can be inoculated at the bottom of the upper organoid culture chip; culture medium is perfused into the lower fluid channel to simulate the real microenvironment of the tumor in vivo;

[0012] The lower fluid channel chip is provided with an anti-collapse column, the anti-collapse column structure is an array micro-column, the diameter of the array micro-column is 1-10mm, the height is 1-10mm, the distance between the array micro-columns is 0.1-10mm, and the height of the array micro-columns is consistent with the height of the lower fluid channel;

[0013] The lower fluid channel chip and the upper organoid culture chip of the microfluidic chip are irreversibly sealed, and the material of the microfluidic chip can be one of polystyrene, polydimethylsiloxane, polymethyl methacrylate, glass and the like;

[0014] The material of the middle porous membrane can be one of polycarbonate, nitrocellulose membrane and the like, and the pore size is 0.4-10 μm;

[0015] The channel inlet of the lower fluid channel chip is connected to a syringe and a syringe pump, the flow rate is set, and the chip is placed in an incubator for perfusion culture. The perfusion culture time is 2-10 days.

[0016] S3: Preparation of biomaterials on the chip: culture the selected tumor cells or tissues and implant them into the corresponding positions of the organoid chip;

[0017] S4: Provide an appropriate physiological environment: In order to simulate the real biological tissue environment, it is necessary to provide appropriate physiological conditions in the organoid chip;

[0018] S5: Monitor and analyze tumor growth and behavior: Use appropriate bioimaging techniques, cell analysis techniques, and biological assays;

[0019] S6: Optimize the model and experimental conditions: According to the experimental results, optimize the model and experimental conditions to improve the authenticity and repeatability of the model;

[0020] S7: Application in research and drug screening: The established tumor tissue models based on organoid chips can be used to study tumor biology, drug toxicity and drug screening.

[0021] Preferably, the S1 is further expressed as:

[0022] S1.1: Tumor type and characteristics: First, the type of tumor to be studied must be clearly defined, as different tumors vary significantly in growth, metastasis, and response to treatment.

[0023] S1.2: Cell line stability and culturability: It is important to select stable, sustainably culturable cell lines to ensure experimental stability and reproducibility;

[0024] S1.3: Patient-derived tumor samples: Use of primary tumor samples from patients can help to more realistically reflect individual differences and patient-specific biological characteristics;

[0025] S1.4: Cell labeling and gene expression: Consider selecting cell lines that are labeled or express specific genes, which can help researchers track cell growth, differentiation, and behavior;

[0026] S1.5: Cellular infiltration and invasiveness: To study the invasion and metastasis characteristics of tumors, it is important to select cell lines or samples with corresponding characteristics;

[0027] S1.6: Selection of control group: In order to establish comparison and control experimental conditions, it is necessary to select some non-tumor cells or normal tissue samples as the control group;

[0028] S1.7: Screening plan: Obtain the patient's tumor tissue during surgery and establish a tumor organoid model in vitro.

[0029] Preferably, S2 is further expressed as:

[0030] S2.1: Chip size and shape: Determine the size and shape of the organoid chip, which should be consistent with the actual tumor tissue being simulated;

[0031] S2.2: Chip materials: Select appropriate materials to construct organoid chips, and these materials should be biocompatible;

[0032] S2.3: Design of the microstructure: The design of the internal microstructure is a critical step because it directly affects the physiology of the model;

[0033] S2.4: Fluid dynamics: Fluid dynamics should be considered during the design process to ensure that media, oxygen, and nutrients can flow efficiently throughout the chip;

[0034] S2.5: Support for multiple cell types: Considering that tumor tissues usually contain multiple cell types, the design of the chip needs to consider supporting the growth and interaction of multiple cell types;

[0035] S2.6: Integrated sensors and monitoring systems: Integrated sensors and monitoring systems to monitor cell growth, metabolic activity, and cell death biological parameters in real time;

[0036] S2.7: Simulation and Optimization: Computer simulations are performed to evaluate the performance of the chip before the chip is actually manufactured;

[0037] S2.8: Personalized treatment: Screening of drug sensitivity of tumor organoid chips can provide clinicians with more effective medication plans.

[0038] Preferably, S3 is further expressed as:

[0039] S3.1: Cell culture: If you choose to use a tumor cell line, you first need to culture the cells, which includes extracting the cells and culturing them in culture medium to ensure stable growth and expansion of the cells;

[0040] S3.2: Cell labeling: In some cases, tumor cells can be labeled using cell labeling techniques, fluorescent labeling or other labeling methods;

[0041] S3.3: Tissue processing: The choice of using patient-derived tumor tissue samples requires appropriate tissue processing;

[0042] S3.4: Cell and tissue implantation: Once the cells or tissues are cultured or processed, the next step is to implant them into the corresponding positions of the organoid chip;

[0043] S3.5: Cell tissue distribution control: During implantation, the distribution of cells or tissues on the chip needs to be considered;

[0044] S3.6: Regulation of culture conditions: Once cells or tissues are implanted on the chip, the culture conditions need to be regulated to provide an appropriate physiological environment;

[0045] S3.7: Stability and bioactivity testing of biomaterials: After implanting cells or tissues on the chip, stability and bioactivity testing are required.

[0046] Preferably, the S4 is further expressed as:

[0047] S4.1: Cell culture medium selection: Selecting an appropriate culture medium is a critical step. The culture medium should contain essential nutrients, such as amino acids, sugars, vitamins, and minerals, to meet the growth and functional requirements of cells.

[0048] S4.2: Temperature control: Maintaining proper temperature is a key factor in maintaining normal physiological functions of cells;

[0049] S4.3: Gas concentration and atmosphere control: In vivo, cells are usually exposed to specific gas concentrations and atmospheres. Mammalian cells are usually cultured in an atmosphere of 5% carbon dioxide.

[0050] S4.4: Humidity control: Proper humidity is essential to maintain water balance in the culture environment;

[0051] S4.5: Microenvironment regulation: Organoid chips should be designed to regulate microenvironmental parameters, such as flow rate, pressure, and oxygen concentration gradient, to better simulate the complex microenvironment of in vivo tissues;

[0052] S4.6: Nutrient supply: Providing appropriate nutrients is necessary to maintain cell survival and function;

[0053] S4.7: pH control: Maintaining proper acid-base balance is essential for normal cell function;

[0054] S4.8: Resuspend cells: Add the cell suspension dropwise to the chip and resuspend the cells with the matrix. The cell resuspended solution can be spotted into the microwells of the chip and placed in a cell culture incubator for 5-10 minutes to allow the matrix to solidify.

[0055] Preferably, the S5 is further expressed as:

[0056] S5.1: Bioimaging technology: Fluorescence microscopy: fluorescent markers are used to label tumor cells or related molecules, and the growth and metastasis process is observed and recorded using a fluorescence microscope;

[0057] S5.2: Cell analysis techniques: Cell counting and viability analysis: Use automated cell counters and viability assays to monitor changes in tumor cell numbers and their biological activity;

[0058] S5.3: Molecular biology detection methods: Gene expression analysis: Use real-time fluorescence quantitative PCR or RNA sequencing technology to study the expression levels of tumor-related genes under different conditions;

[0059] S5.4: Microenvironment simulation and assessment of therapeutic response: Simulating the in vivo microenvironment: Modeling the tumor microenvironment in organoid chips, including cellular interactions, extracellular matrix, and vascular supply, to more realistically replicate the conditions in which tumors grow;

[0060] S5.5: Real-time monitoring and automation: Real-time monitoring system: Integrate sensors and real-time monitoring systems to ensure real-time tracking of tumor growth and behavior;

[0061] S5.6: Genetic testing: For patients with positive genetic test results, gene editing technology is used for in vitro targeted verification.

[0062] Preferably, the S6 is further expressed as:

[0063] S6.1: Optimization of cell culture conditions: Medium composition: Evaluate the concentration of various components in the cell culture medium to ensure that appropriate nutrients and growth factors are provided to promote normal growth of tumor cells on the chip;

[0064] S6.2: Optimization of chip design: cell layout and arrangement: Adjust the layout and arrangement of cells on the chip to better mimic the structure of tumor tissue in vivo, such as simulating the heterogeneity of the tumor edge and center;

[0065] S6.3: Improvement of experimental operation procedures: Standardized experimental procedures: Establish standard experimental operation procedures to ensure that each experiment is performed in a similar manner to improve repeatability; Automated experimental operation: Consider introducing an automated system to reduce human errors and improve the accuracy and consistency of the experiment;

[0066] S6.4: Improvements in data analysis and interpretation: Standardized data collection: Use uniform data collection formats and standards to make it easier to compare and analyze results from different experiments;

[0067] S6.5: Validation and reproducibility of experimental results: Validation of experimental results: Conduct validation experiments to confirm previous results and ensure the reliability of experimental results;

[0068] S6.6: Clinical trials: Clinicians recommend other patients for tumor drug sensitivity testing, auxiliary precision treatment, preoperative neoadjuvant, postoperative treatment, and tumor patients who cannot undergo surgery.

[0069] Preferably, the S7 is further expressed as:

[0070] S7.1: Tumor biology research: Tumor microenvironment simulation: Organoid chips can simulate the tumor microenvironment, including the interaction between tumor cells and surrounding cells and matrix, which is helpful for studying the development, growth and metastasis of tumors;

[0071] S7.2: Drug toxicity assessment: Toxicity screening: Drug toxicity assessment using organoid chip models can more accurately simulate the physiological responses of the human body, thereby improving the accuracy of predicting toxicity;

[0072] S7.3: Drug screening and development: Pharmacodynamic studies: Utilize organoid chip models to conduct pharmacodynamic studies to evaluate the efficacy and dose-response relationship of candidate drugs and accelerate the drug screening process;

[0073] S7.4: Mechanism analysis and new drug discovery: Disease mechanism research: Organoid chip models can be used to deeply explore the pathogenesis of tumors and discover new therapeutic targets and drugs;

[0074] S7.5: Data integration and multi-omics analysis: Multi-omics data integration: combining experimental data from organoid chip models;

[0075] S7.6: Accelerate the process: Accelerate the process of personalized medicine, reduce medical resources, and reduce the family burden of patients.

[0076] The present invention has the following beneficial effects:

[0077] 1. The present invention selects suitable tumor cell lines or tumor tissue samples obtained from patients, and designs the structure of the organoid chip based on existing tumor biology knowledge and engineering principles to ensure the stability and repeatability of the experiment, which helps to more truly reflect individual differences and patient-specific biological characteristics.

[0078] 2. The present invention simulates the real biological tissue environment by culturing and implanting the selected tumor cells or tissues into the corresponding positions of the organoid chip to ensure the stable growth and expansion of the cells. During the implantation process, the distribution of cells or tissues on the chip needs to be considered to provide an appropriate physiological environment.

[0079] 3. The present invention optimizes the model and experimental conditions according to the experimental results by using appropriate bio-imaging technology, cell analysis technology and biological detection methods to improve the authenticity and repeatability of the model, so as to more realistically replicate the conditions for tumor growth and ensure real-time tracking of tumor growth and behavior.

[0080] Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for describing the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0082] Figure 1 is a flowchart of the present invention;

[0083] Figure 2The chip design diagram of the present invention, a is the lower fluid channel chip, b is the middle porous membrane layer, and c is the upper organoid culture chip;

[0084] Figure 3 It is a physical diagram of the present invention;

[0085] Figure 4 This is a bright field image of cells on the chip. DETAILED DESCRIPTION

[0086] 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.

[0087] See also Figure 1-4 The present invention provides a method for establishing a tumor tissue model based on an organoid chip, comprising the following steps:

[0088] S1: cells or tissues are used to build models on organoid chips;

[0089] S2: To determine the size, shape, material, and internal microstructure layout of the chip;

[0090] S3: It is convenient to culture tumor cells or tissues and implant them into the corresponding positions of the organoid chip;

[0091] S4: convenient for cell culture;

[0092] S5: To monitor and analyze the growth, metastasis, and treatment response of tumor tissues simulated on organoid chips;

[0093] S6: To improve the experimental operation process and other aspects;

[0094] S7: It is convenient for studying areas such as tumor biology, drug toxicity and drug screening.

[0095] S1 is further expressed as:

[0096] S1.1: To ensure the biological relevance of the model;

[0097] S1.2: To ensure the stability and repeatability of the experiment;

[0098] S1.3: Helps to more realistically reflect individual differences and patient-specific biological characteristics;

[0099] S1.4: Helps researchers track cell growth, differentiation, and behavior;

[0100] S1.5: Helps to better simulate the behavior of actual tumors in vivo;

[0101] S1.6: This helps identify tumor-specific changes;

[0102] S1.7: Facilitates rapid screening of the best tumor treatment recommendations.

[0103] Select appropriate cell lines or tissue sources: The above steps involve the selection of cell lines or tissue samples, which can ensure that the established tumor tissue model based on the organoid chip is biologically accurate, reliable and practical.

[0104] S2 is further expressed as:

[0105] S2.1: Helps determine the size and shape of the organoid chip;

[0106] S2.2: Facilitates the simulation of the physical and chemical properties of tumor tissue;

[0107] S2.3: In order to simulate the complex structure of tumor tissue;

[0108] S2.4: helps provide more realistic physiological conditions;

[0109] S2.5: Facilitates the growth and interaction of multiple cell types;

[0110] S2.6: It helps researchers obtain detailed experimental data;

[0111] S2.7: Ensure that the chip can provide appropriate physiological conditions and experimental effects;

[0112] S2.8: In order to provide personalized treatment for cancer patients.

[0113] Designing the organoid chip: It is critical to work closely with professionals in biology and engineering to ensure that the chip design not only meets the requirements of tumor biology but is also robust and feasible in experiments.

[0114] S3 is further expressed as:

[0115] S3.1: To ensure the biological activity and stability of cells;

[0116] S3.2: helps track the location, growth, and behavior of cells in the model;

[0117] S3.3: to extract single cells or cell clusters;

[0118] S3.4: It is convenient to implant them into the corresponding positions of the organoid chip;

[0119] S3.5: Ensure uniform distribution or specific arrangement of cells or tissues on the chip;

[0120] S3.6: Facilitate the optimization of culture medium formulation, temperature, humidity, atmosphere and other parameters;

[0121] S3.7: To ensure the survival and functionality of cells or tissues on the chip.

[0122] Preparation of biomaterials on chip: The above steps should be performed aseptically and in accordance with laboratory safety regulations to ensure the accuracy and reliability of the experimental results. In addition, each step in the preparation process should be recorded and documented for subsequent analysis and optimization.

[0123] S4 is further expressed as:

[0124] S4.1: To simulate its in vivo growth environment;

[0125] S4.2: To ensure that cells are grown at a temperature similar to the in vivo environment;

[0126] S4.3: To simulate in vivo conditions;

[0127] S4.4: Helps prevent water loss in cells and tissues and maintains the stability of the culture environment;

[0128] S4.5: To better simulate the complex microenvironment of in vivo tissues;

[0129] S4.6: To simulate the nutritional conditions required by cells in vivo;

[0130] S4.7: Ensure that the acid-base balance of the culture medium is within the ideal range;

[0131] S4.8: To prevent excessive evaporation of the culture medium.

[0132] Providing an appropriate physiological environment: The above steps can better simulate the physiological environment in vivo by carefully regulating these factors, providing a more realistic and reliable model for organoid chips, which is helpful for biological research and drug testing.

[0133] S5 is further expressed as:

[0134] S5.1: To obtain more comprehensive information about the tumor structure;

[0135] S5.2: To understand its growth and treatment response;

[0136] S5.3: helps understand its growth and treatment response;

[0137] S5.4: To more realistically replicate the conditions under which tumors grow;

[0138] S5.5: Facilitates the extraction of key information about the biological behavior of tumors;

[0139] S5.6: It is convenient to assist in formulating appropriate targeted treatment plans.

[0140] Monitoring and analyzing tumor growth and behavior: By combining these technologies and methods, the above steps can provide a more comprehensive and accurate understanding of the growth, metastasis and treatment response of tumor tissues simulated on organoid chips, providing key information for tumor biology and drug development.

[0141] S6 is further expressed as:

[0142] S6.1: To better simulate the in vivo situation;

[0143] S6.2: To better simulate the tumor microenvironment;

[0144] S6.3: Ensure the accuracy and reliability of experimental results;

[0145] S6.4: to extract deeper information and discover potential connections and patterns;

[0146] S6.5: To facilitate the replication and verification of experimental results;

[0147] S6.6: Facilitates the screening of clinical drug regimens across cancer types.

[0148] Optimize the model and experimental conditions: Through these optimization steps, the above steps can improve the authenticity and repeatability of the model, thereby enhancing the scientific reliability of the research and the feasibility of practical application.

[0149] S7 is further expressed as:

[0150] S7.1: To better understand tumor heterogeneity and its impact on treatment response;

[0151] S7.2: Facilitate the assessment of drug toxicity in different organs and predict the extent of systemic toxicity;

[0152] S7.3: Facilitate personalized drug testing and guide the formulation of clinical treatment plans;

[0153] S7.4: Contribute to the discovery of new drugs or drug combinations with potential therapeutic effects;

[0154] S7.5: It is helpful for comprehensive analysis to reveal the combined effects of drug mechanism of action and treatment response;

[0155] S7.6: To assist in formulating subsequent treatment plans.

[0156] Tumor organoid culture

[0157] Place the chip in a clean bench and sterilize it with ultraviolet light. First, use a pipette to add tumor organoid culture medium to the fluid channel of the lower culture chip. Then, place 5 microliters of Matrigel resuspended with tumor organoids in the upper culture chamber of the chip. Then, place the chip in a 37-degree Celsius incubator for 10 minutes to allow Matrigel to solidify. After 10 minutes, fill the upper culture chamber of the chip with culture medium. Finally, connect the perfusion tube and syringe to the inlet side of the lower culture channel, use a perfusion pump to perfuse the fluid, and place it in a 37-degree Celsius incubator for culture. Figure 3 shown.

[0158] Application in research and drug screening: By further refining the above steps in these aspects, the organoid chip model will provide more accurate and efficient tools and platforms for tumor treatment and drug development, which is expected to promote progress in the field of tumor research and accelerate the discovery and clinical application of new drugs.

[0159] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0160] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A method for establishing a tumor tissue model based on an organoid chip, characterized in that: The following steps are involved: S1: Select appropriate cell lines or tissue sources: First, you need to select appropriate tumor cell lines or tumor tissue samples obtained from patients; S2: Design organoid chips: Based on existing knowledge of tumor biology and engineering principles, design tumor organoid chips that mimic the tumor microenvironment; S3: Preparation of biomaterials on the chip: culture the selected tumor cells or tissues and implant them into the corresponding positions of the organoid chip; S4: Provide an appropriate physiological environment: In order to simulate the real biological tissue environment, it is necessary to provide appropriate physiological conditions in the organoid chip; S5: Monitor and analyze tumor growth and behavior: Use appropriate bioimaging techniques, cell analysis techniques, and biological assays; S6: Optimize the model and experimental conditions: According to the experimental results, optimize the model and experimental conditions to improve the authenticity and repeatability of the model; S7: Application in research and drug screening: The established tumor tissue models based on organoid chips can be used to study tumor biology, drug toxicity and drug screening.

2. The method for establishing a tumor tissue model based on an organoid chip according to claim 1, characterized in that: The S1 is further expressed as: S1.1: Tumor type and characteristics: First, the type of tumor to be studied must be clearly defined, as different tumors vary significantly in growth, metastasis, and response to treatment. S1.2: Cell line stability and culturability: It is important to select stable, sustainably culturable cell lines to ensure experimental stability and reproducibility; S1.3: Patient-derived tumor samples: Use of primary tumor samples from patients can help to more realistically reflect individual differences and patient-specific biological characteristics; S1.4: Cell labeling and gene expression: Consider selecting cell lines that are labeled or express specific genes, which can help researchers track cell growth, differentiation, and behavior; S1.5: Cellular infiltration and invasiveness: To study the invasion and metastasis characteristics of tumors, it is important to select cell lines or samples with corresponding characteristics; S1.6: Selection of control group: In order to establish comparison and control experimental conditions, it is necessary to select some non-tumor cells or normal tissue samples as the control group; S1.7: Screening plan: Obtain the patient's tumor tissue during surgery and establish a tumor organoid model in vitro.

3. The method for establishing a tumor tissue model based on an organoid chip according to claim 2, characterized in that: The S2 is further expressed as: S2.1: Chip size and shape: Determine the size and shape of the organoid chip, which should be consistent with the actual tumor tissue being simulated; S2.2: Chip materials: Select appropriate materials to construct organoid chips, and these materials should be biocompatible; S2.3: Design of the microstructure: The design of the internal microstructure is a critical step because it directly affects the physiology of the model; S2.4: Fluid dynamics: Fluid dynamics should be considered during the design process to ensure that media, oxygen, and nutrients can flow efficiently throughout the chip; S2.5: Support for multiple cell types: Considering that tumor tissues usually contain multiple cell types, the design of the chip needs to consider supporting the growth and interaction of multiple cell types; S2.6: Integrated sensors and monitoring systems: Integrated sensors and monitoring systems to monitor cell growth, metabolic activity, and cell death biological parameters in real time; S2.7: Simulation and Optimization: Computer simulations are performed to evaluate the performance of the chip before the chip is actually manufactured; S2.8: Personalized treatment: Screening of drug sensitivity of tumor organoid chips can provide clinicians with more effective medication plans.

4. The method for establishing a tumor tissue model based on an organoid chip according to claim 3, characterized in that: The S3 is further expressed as: S3.1: Cell culture: If you choose to use a tumor cell line, you first need to culture the cells, which includes extracting the cells and culturing them in culture medium to ensure stable growth and expansion of the cells; S3.2: Cell labeling: In some cases, tumor cells can be labeled using cell labeling techniques, fluorescent labeling or other labeling methods; S3.3: Tissue processing: The choice of using patient-derived tumor tissue samples requires appropriate tissue processing; S3.4: Cell and tissue implantation: Once the cells or tissues are cultured or processed, the next step is to implant them into the corresponding positions of the organoid chip; S3.5: Cell tissue distribution control: During implantation, the distribution of cells or tissues on the chip needs to be considered; S3.6: Regulation of culture conditions: Once cells or tissues are implanted on the chip, the culture conditions need to be regulated to provide an appropriate physiological environment; S3.7: Stability and bioactivity testing of biomaterials: After implanting cells or tissues on the chip, stability and bioactivity testing are performed.

5. The method for establishing a tumor tissue model based on an organoid chip according to claim 4, characterized in that: The S4 is further expressed as: S4.1: Cell culture medium selection: Selecting an appropriate culture medium is a critical step. The culture medium should contain essential nutrients, such as amino acids, sugars, vitamins, and minerals, to meet the growth and functional requirements of cells. S4.2: Temperature control: Maintaining proper temperature is a key factor in maintaining normal physiological functions of cells; S4.3: Gas concentration and atmosphere control: In vivo, cells are usually exposed to specific gas concentrations and atmospheres. Mammalian cells are usually cultured in an atmosphere of 5% carbon dioxide. S4.4: Humidity control: Proper humidity is essential to maintain water balance in the culture environment; S4.5: Microenvironment regulation: Organoid chips should be designed to regulate microenvironmental parameters, such as flow rate, pressure, and oxygen concentration gradient, to better simulate the complex microenvironment of in vivo tissues; S4.6: Nutrient supply: Providing appropriate nutrients is necessary to maintain cell survival and function; S4.7: pH control: Maintaining proper acid-base balance is essential for normal cell function; S4.8: Resuspend cells: Add the cell suspension dropwise to the chip and resuspend the cells with the matrix. The cell resuspended solution can be spotted into the microwells of the chip and placed in a cell culture incubator for 5-10 minutes to allow the matrix to solidify.

6. The method for establishing a tumor tissue model based on an organoid chip according to claim 5, characterized in that: The S5 is further expressed as: S5.1: Bioimaging technology: Fluorescence microscopy: fluorescent markers are used to label tumor cells or related molecules, and the growth and metastasis process is observed and recorded using a fluorescence microscope; S5.2: Cell analysis techniques: Cell counting and viability analysis: Use automated cell counters and viability assays to monitor changes in tumor cell numbers and their biological activity; S5.3: Molecular biology detection methods: Gene expression analysis: Use real-time fluorescence quantitative PCR or RNA sequencing technology to study the expression levels of tumor-related genes under different conditions; S5.4: Microenvironment simulation and assessment of therapeutic response: Simulating the in vivo microenvironment: Modeling the tumor microenvironment in organoid chips, including cellular interactions, extracellular matrix, and vascular supply, to more realistically replicate the conditions in which tumors grow; S5.5: Real-time monitoring and automation: Real-time monitoring system: Integrate sensors and real-time monitoring systems to ensure real-time tracking of tumor growth and behavior; S5.6: Genetic testing: For patients with positive genetic test results, gene editing technology is used for in vitro targeted verification.

7. The method for establishing a tumor tissue model based on an organoid chip according to claim 6, characterized in that: The S6 is further expressed as: S6.1: Optimization of cell culture conditions: Medium composition: Evaluate the concentration of various components in the cell culture medium to ensure that appropriate nutrients and growth factors are provided to promote normal growth of tumor cells on the chip; S6.2: Optimization of chip design: cell layout and arrangement: Adjust the layout and arrangement of cells on the chip to better mimic the structure of tumor tissue in vivo, such as simulating the heterogeneity of the tumor edge and center; S6.3: Improvement of experimental operation procedures: Standardized experimental procedures: Establish standard experimental operation procedures to ensure that each experiment is performed in a similar manner to improve repeatability; Automated experimental operation: Consider introducing an automated system to reduce human errors and improve the accuracy and consistency of the experiment; S6.4: Improvements in data analysis and interpretation: Standardized data collection: Use uniform data collection formats and standards to make it easier to compare and analyze results from different experiments; S6.5: Validation and reproducibility of experimental results: Validation of experimental results: Conduct validation experiments to confirm previous results and ensure the reliability of experimental results; S6.6: Clinical trials: Clinicians recommend other patients for tumor drug sensitivity testing, auxiliary precision treatment, preoperative neoadjuvant, postoperative treatment, and tumor patients who cannot undergo surgery.

8. The method for establishing a tumor tissue model based on an organoid chip according to claim 7, characterized in that: The S7 is further expressed as: S7.1: Tumor biology research: Tumor microenvironment simulation: Organoid chips can simulate the tumor microenvironment, including the interaction between tumor cells and surrounding cells and matrix, which is helpful for studying the development, growth and metastasis of tumors; S7.2: Drug toxicity assessment: Toxicity screening: Drug toxicity assessment using organoid chip models can more accurately simulate the physiological responses of the human body, thereby improving the accuracy of predicting toxicity; S7.3: Drug screening and development: Pharmacodynamic studies: Utilize organoid chip models to conduct pharmacodynamic studies to evaluate the efficacy and dose-response relationship of candidate drugs and accelerate the drug screening process; S7.4: Mechanism analysis and new drug discovery: Disease mechanism research: Organoid chip models can be used to deeply explore the pathogenesis of tumors and discover new therapeutic targets and drugs; S7.5: Data integration and multi-omics analysis: Multi-omics data integration: combining experimental data from organoid chip models; S7.6: Accelerate the process: Accelerate the process of personalized medicine, reduce medical resources, and reduce the family burden of patients.

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