Tumor organ chip and application thereof in tumor organ culture

Abstract

The invention discloses a tumor organoid chip and application thereof in tumor organoid culture, the tumor organoid chip comprises a plurality of microfluidic units, the microfluidic units are used for simulating different tumor microenvironments, each microfluidic unit comprises a plurality of microchannels, an inlet and outlet channel and a microfluidic system, the conveying device is used for accurately conveying culture media, drugs, cell factors and growth factors; the 3D culture module adopts a nano-scale porous material and a biodegradable bracket to simulate a three-dimensional structure of a tumor tissue, and due to the design of the 3D culture module, the tumor-like organs can be self-assembled into a multi-layer tissue structure similar to a human tumor. By introducing a multi-cell co-culture system and a microfluidic technology, the tumor organ-like chip can simulate complex in-vivo microenvironments, including immune escape, angiogenesis, inter-tumor heterogeneity and the like, of tumors. Interaction among different cell types can be accurately controlled in the chip, so that tumor organs are closer to the physiological state of human tumors.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and specifically to a tumor organoid chip and its application in tumor organoid culture. Background Art

[0002] Tumor organoid chips are an advanced platform that integrates microfluidics technology and an organoid culture system. They can simulate the tumor microenvironment in vitro and reproduce the complex structure and function of tumor tissue. By precisely controlling cell growth and regulating the microenvironment, this technology cultivates three-dimensional tumor organoids with cancer characteristics, which researchers can use for tumor biology research, drug screening, and treatment response evaluation. Tumor organoid chips not only provide a model that is closer to real human tumors, but also can evaluate the efficacy and toxicity of drugs in a shorter period of time, providing important support for personalized treatment and precision medicine. The application of this technology is expected to improve the efficiency of tumor research, accelerate the development of new drugs, and promote innovation in cancer treatment.

[0003] In existing technologies, tumor organoid chips and their applications in tumor organoid culture have many advantages, but they also have some disadvantages and challenges, mainly including the following aspects:

[0004] Model complexity and reproducibility issues:

[0005] The culturing process of tumor organoids is complex and requires precise microenvironmental control. Different cell sources, culture conditions, and chip designs can lead to differences in tumor organoid morphology and function, thus affecting the reproducibility of experimental results. This means that standardization of tumor organoid models remains a challenge.

[0006] Technical limitations of the culture system:

[0007] While microfluidics can effectively simulate the tumor microenvironment, existing tumor organoid chips often have technical limitations. For example, they cannot perfectly replicate the complex physiological processes in the human body, such as immune responses and blood flow. In addition, tumor organoids are small in size and may not fully reflect the characteristics of large-scale tumors.

[0008] Lack of intertissue heterogeneity:

[0009] Tumors are inherently highly heterogeneous, comprising multiple cell types and their interactions. Current tumor organoid chips primarily focus on a single tumor type or a limited number of cell types, potentially failing to fully simulate the heterogeneity between or within tumors. This poses challenges for their application in precision medicine and personalized treatment.

[0010] High cost and cumbersome operation:

[0011] The fabrication, cultivation, and operation of tumor organoid chips require advanced equipment and specialized techniques, increasing experimental costs and operational complexity. Chip design and material selection also directly impact performance, and reducing costs while improving usability remains a pressing challenge.

[0012] The model has limited physiological relevance:

[0013] Although tumor organoid chips can reproduce the biological characteristics of tumors to a certain extent, their complete similarity to human tumors is still insufficient, especially in long-term culture and the reproduction of complex disease models (such as metastasis and immune response). This limits their accuracy and effectiveness in clinical applications.

[0014] Limitations of drug screening:

[0015] While tumor organoid chips can be used for preliminary drug screening, their ability to effectively predict clinical treatment responses is limited. Different tumor types and individual differences can lead to differences in drug response, so further optimization of the model and drug screening process is needed.

[0016] To this end, we propose a tumor organoid chip and its application in tumor organoid culture. Summary of the Invention

[0017] To achieve the above objectives, the present invention provides the following technical solution: a tumor organ-on-a-chip, comprising:

[0018] Multiple microfluidic units, each of which is used to simulate different tumor microenvironments. The microfluidic units include multiple microchannels, inlet and outlet channels, and a microfluidic system for accurately delivering culture media, drugs, cytokines, and growth factors.

[0019] A 3D culture module, which uses nanoporous materials and biodegradable scaffolds to simulate the three-dimensional structure of tumor tissue. The design of the 3D culture module enables tumor organoids to self-assemble into a multi-layered tissue structure similar to that of human tumors;

[0020] A multi-cell co-culture system comprising tumor cells, immune cells, fibroblasts, and other related cells, for reproducing tumor heterogeneity;

[0021] A controllable environmental monitoring and feedback system includes a temperature sensor, a pH sensor, an oxygen concentration sensor, a cell metabolism monitor, etc., which is used to monitor environmental changes during the culture process in real time and automatically adjust parameters in the microchannel.

[0022] Preferably, the microfluidic unit adjusts the flow rate and direction of the fluid through microfluidic technology to achieve precise control of the tumor microenvironment, thereby simulating different tumor development stages, different treatment plans and different immune responses.

[0023] Preferably, the nanoscale porous material in the 3D culture module is a biocompatible material, such as polydimethylsiloxane (PDMS), polyurethane (PU), etc., which has high cell adhesion and biodegradability and supports the growth and differentiation of tumor cells.

[0024] Preferably, the multi-cell co-culture system supports the co-culture of tumor cells with immune cells, fibroblasts, endothelial cells and other cells to simulate complex biological processes such as immune escape and angiogenesis in the tumor microenvironment.

[0025] Preferably, the controllable environment monitoring and feedback system automatically adjusts the fluid flow rate, temperature, pH value and oxygen concentration in the microfluidic system based on sensor detection data to maintain the growth of tumor organoids under optimal culture conditions.

[0026] A method for applying a tumor organoid chip comprises the following steps:

[0027] S1 adds patient-derived tumor cells, immune cells, fibroblasts and other cells into the multi-cell co-culture system of the chip in a predetermined ratio;

[0028] S2 precisely controls the delivery of culture medium, drugs, and cytokines through microfluidic units within the chip to simulate tumor growth and immune response;

[0029] S3 uses a controlled environment monitoring and feedback system to monitor parameters such as temperature, pH, and oxygen concentration during the culture process in real time and adjust the culture conditions;

[0030] S4 conducts drug screening and efficacy evaluation, analyzing the effects of different drugs on the growth, metastasis and immune escape of tumor organoids.

[0031] Compared with the prior art, the present invention provides a tumor organoid chip and its application in tumor organoid culture, which has the following beneficial effects:

[0032] 1. This tumor organoid chip and its application in tumor organoid culture, through the introduction of a multi-cell co-culture system and microfluidics technology, can simulate the complex microenvironment of tumors in the body, including immune escape, angiogenesis, and inter-tumor heterogeneity. The chip can precisely control the interactions between different cell types, making the tumor organoids more similar to the physiological state of human tumors.

[0033] 2. The tumor organoid chip and its application in tumor organoid culture. The chip's modular design and precise environmental control make the culture process more controllable. Culture conditions such as temperature, pH, and oxygen concentration can be monitored and automatically adjusted in real time, ensuring experimental reproducibility and standardization. The integration of multiple biomarkers and sensors also achieves batch-to-batch consistency, reducing experimental errors.

[0034] 3. This tumor organoid chip and its application in tumor organoid culture, through a high-throughput drug screening system, allows researchers to quickly test the efficacy of multiple drugs and assess their effects on tumor organoid growth, metastasis, and immune response. Compared to traditional two-dimensional culture or animal models, the chip of this invention can greatly improve the efficiency of drug screening.

[0035] 4. The tumor organ-on-chip and its application in tumor organoid culture are different from single cell culture systems. The multi-cell co-culture system of the present invention can simultaneously simulate the interaction between tumor cells and immune cells, fibroblasts, etc., providing a more realistic immune microenvironment and effectively evaluating the effects of immunotherapy or immune escape mechanisms on tumors.

[0036] 5. This tumor organoid chip and its application in tumor organoid culture can use patient-derived tumor cells for personalized drug screening, supporting precision medicine. By accurately simulating patient tumor models, it can provide more targeted treatment options for clinical treatment, thereby improving treatment efficacy and reducing side effects. DETAILED DESCRIPTION

[0037] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0038] Example

[0039] An embodiment of a tumor organoid chip and its application in tumor organoid culture

[0040] A tumor organoid chip, comprising:

[0041] Multiple microfluidic units, each of which is used to simulate different tumor microenvironments. The microfluidic units include multiple microchannels, inlet and outlet channels, and a microfluidic system for accurately delivering culture media, drugs, cytokines, and growth factors.

[0042] A 3D culture module, which uses nanoporous materials and biodegradable scaffolds to simulate the three-dimensional structure of tumor tissue. The design of the 3D culture module enables tumor organoids to self-assemble into a multi-layered tissue structure similar to that of human tumors;

[0043] A multi-cell co-culture system comprising tumor cells, immune cells, fibroblasts, and other related cells, for reproducing tumor heterogeneity;

[0044] A controllable environmental monitoring and feedback system includes a temperature sensor, a pH sensor, an oxygen concentration sensor, a cell metabolism monitor, etc., which is used to monitor environmental changes during the culture process in real time and automatically adjust parameters in the microchannel.

[0045] Specifically, the microfluidic unit uses microfluidic technology to regulate the flow rate and direction of the fluid to achieve precise control of the tumor microenvironment, thereby simulating different tumor development stages, different treatment plans and different immune responses.

[0046] Specifically, the nanoscale porous materials in the 3D culture module are biocompatible materials, such as polydimethylsiloxane (PDMS), polyurethane (PU), etc., which have high cell adhesion and biodegradability, and support the growth and differentiation of tumor cells.

[0047] Specifically, the multi-cell co-culture system supports the co-culture of tumor cells with immune cells, fibroblasts, endothelial cells and other cells to simulate complex biological processes such as immune escape and angiogenesis in the tumor microenvironment.

[0048] Specifically, the controllable environment monitoring and feedback system automatically adjusts the fluid flow rate, temperature, pH value and oxygen concentration in the microfluidic system based on sensor detection data to maintain the growth of tumor organoids under optimal culture conditions.

[0049] A method for applying a tumor organoid chip comprises the following steps:

[0050] S1 adds patient-derived tumor cells, immune cells, fibroblasts and other cells into the multi-cell co-culture system of the chip in a predetermined ratio;

[0051] S2 precisely controls the delivery of culture medium, drugs, and cytokines through microfluidic units within the chip to simulate tumor growth and immune response;

[0052] S3 uses a controlled environment monitoring and feedback system to monitor parameters such as temperature, pH, and oxygen concentration during the culture process in real time and adjust the culture conditions;

[0053] S4 conducts drug screening and efficacy evaluation, analyzing the effects of different drugs on the growth, metastasis and immune escape of tumor organoids.

[0054] Through the above technical solutions, the present invention, by incorporating a multicellular co-culture system and microfluidics, enables tumor organoids to simulate the complex in vivo tumor microenvironment, including immune evasion, angiogenesis, and inter-tumor heterogeneity. The chip precisely controls the interactions between different cell types, making tumor organoids more closely resemble the physiological state of human tumors. The chip's modular design and precise environmental control make the culture process more controllable. Culture conditions such as temperature, pH, and oxygen concentration can be monitored and automatically adjusted in real time, ensuring experimental reproducibility and standardization. The integration of multiple biomarkers and sensors also achieves batch-to-batch consistency, reducing experimental error. Through a high-throughput drug screening system, researchers can rapidly test the efficacy of multiple drugs and evaluate their effects on tumor organoid growth, metastasis, and immune responses. Compared to traditional two-dimensional culture or animal models, the chip of the present invention significantly improves the efficiency of drug screening. Unlike single-cell culture systems, the multicellular co-culture system of the present invention can simultaneously simulate the interactions between tumor cells and immune cells, such as fibroblasts, providing a more realistic immune microenvironment and effectively evaluating the effects of immunotherapy or immune escape mechanisms on tumors. This chip can be used to screen personalized drugs using patient-derived tumor cells, supporting precision medicine. By accurately simulating patient tumor models, it can provide more targeted treatment options for clinical treatment, thereby improving treatment efficacy and reducing side effects.

[0055] Chip design and manufacturing:

[0056] Microfluidic Unit: The chip's microfluidic unit consists of multiple microchannels, each with micron-scale width and height. These channels control the flow rate of culture media, drugs, cytokines, and other substances. By controlling the flow of fluids, these channels simulate blood flow and drug delivery during tumor growth. Current sensors, pH sensors, and oxygen sensors are built into the channels for real-time monitoring and adjustment of environmental conditions.

[0057] 3D Culture Module: Utilizing biodegradable nanoporous materials (such as polyurethane and polydimethylsiloxane) as scaffolds, the scaffold's pore size and structural design mimic the cell growth environment found in tumor tissue. The nanomaterials promote cell attachment and support the three-dimensional growth of tumor organoids. The scaffold's degradability ensures that cell proliferation is not affected during the culture process.

[0058] Multi-cell co-culture module: The chip is equipped with multiple culture chambers, capable of culturing different cell types separately. During the culture process, tumor cells coexist with immune cells, fibroblasts, endothelial cells, etc. in the same microenvironment, simulating a real tumor microenvironment and immune response.

[0059] Cell treatment and culture:

[0060] Cell source: Tumor cells, immune cells (such as T cells, macrophages), fibroblasts, endothelial cells, etc. are extracted from the patient's body and processed through tissue culture or single-cell separation technology to obtain high-quality cells.

[0061] Cell co-culture: Different cell types are added proportionally to the co-culture chamber in the chip. Tumor cells interact with immune cells, fibroblasts, etc. to form physiologically relevant tumor organoids within the chip.

[0062] Environmental control and monitoring:

[0063] Temperature and pH control: Use temperature and pH sensors to monitor the culture environment and automatically adjust the temperature and pH of the culture medium to ensure that cells grow under optimal conditions.

[0064] Oxygen concentration regulation: The oxygen concentration is detected in real time through an oxygen sensor, and the oxygen concentration in the flow channel is automatically adjusted through a microfluidic unit to simulate the hypoxic environment of the tumor.

[0065] Real-time data monitoring and feedback: All sensor data is fed back to the central control system in real time, and the culture conditions are automatically adjusted through software algorithms to achieve precise culture environment control.

[0066] Drug screening and evaluation:

[0067] Drug input: Through the microfluidic system, drugs of different concentrations (such as chemotherapy drugs, targeted drugs, immune drugs, etc.) are accurately input into the culture chamber to simulate different drug treatment plans.

[0068] Efficacy evaluation: Evaluate the effects of drugs on tumor organoids through cell growth curves, morphological observations, immune labeling, and other means. Use sensors to monitor biological processes such as cell metabolism, proliferation, and migration to assess drug efficacy and toxicity. While the embodiments of the present invention have been shown and described, those skilled in the art will appreciate that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention. The scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A tumor organ-on-a-chip, characterized in that: include: Multiple microfluidic units, each of which is used to simulate different tumor microenvironments. The microfluidic units include multiple microchannels, inlet and outlet channels, and a microfluidic system for accurately delivering culture media, drugs, cytokines, and growth factors. A 3D culture module, which uses nanoporous materials and biodegradable scaffolds to simulate the three-dimensional structure of tumor tissue. The design of the 3D culture module enables tumor organoids to self-assemble into a multi-layered tissue structure similar to that of human tumors; A multi-cell co-culture system comprising tumor cells, immune cells, fibroblasts, and other related cells, for reproducing tumor heterogeneity; A controllable environmental monitoring and feedback system includes a temperature sensor, a pH sensor, an oxygen concentration sensor, a cell metabolism monitor, etc., which is used to monitor environmental changes during the culture process in real time and automatically adjust parameters in the microfluidic channel.

2. The tumor organ-on-a-chip according to claim 1, wherein: The microfluidic unit adjusts the flow rate and direction of the fluid through microfluidic technology to achieve precise control of the tumor microenvironment, thereby simulating different tumor development stages, different treatment plans and different immune responses.

3. The tumor organ-on-a-chip according to claim 1, wherein: The nanoscale porous material in the 3D culture module is a biocompatible material, such as polydimethylsiloxane (PDMS), polyurethane (PU), etc., which has high cell adhesion and biodegradability and supports the growth and differentiation of tumor cells.

4. The tumor organ-on-a-chip according to claim 1, wherein: The multi-cell co-culture system supports the co-culture of tumor cells with immune cells, fibroblasts, endothelial cells and other cells to simulate complex biological processes such as immune escape and angiogenesis in the tumor microenvironment.

5. The tumor organ-on-a-chip according to claim 1, wherein: The controllable environment monitoring and feedback system automatically adjusts the fluid flow rate, temperature, pH value and oxygen concentration in the microfluidic system based on sensor detection data to maintain the growth of tumor organoids under optimal culture conditions.

6. A method for applying a tumor organoid chip, characterized in that: The following steps are involved: S1 adds patient-derived tumor cells, immune cells, fibroblasts and other cells into the multi-cell co-culture system of the chip in a predetermined ratio; S2 precisely controls the delivery of culture medium, drugs, and cytokines through microfluidic units within the chip to simulate tumor growth and immune response; S3 uses a controlled environment monitoring and feedback system to monitor parameters such as temperature, pH, and oxygen concentration during the culture process in real time and adjust the culture conditions; S4 conducts drug screening and efficacy evaluation, analyzing the effects of different drugs on the growth, metastasis and immune escape of tumor organoids.