A colorectal cancer organoid chip

Abstract

The utility model relates to tumor in vitro model technical field discloses a kind of colorectal cancer organoid chip, including chip substrate and composite function layer;Composite function layer is set on chip substrate and is connected with microfluidic regulation and control system and detection module, it is successively support layer, vascularization layer, tumor microenvironment simulation layer and epithelial barrier layer from below to above, each layer is connected by nanofiber bridging structure connection.The utility model restores colorectal cancer tissue hierarchical characteristics by layered structure design, realizes interlayer material exchange and signal conduction using nanofiber bridging structure, in combination with the optical advantage of glass substrate and the cooperation with regulation and control, detection system, colorectal cancer microenvironment and multisystem interaction can be accurately simulated, the problem that existing chip simulation is not comprehensive is solved, research reliability and conversion value are improved, suitable for colorectal cancer basic research and drug development.

Description

Technical Field

[0001] This utility model relates to the field of in vitro tumor model technology, and more specifically, to an organoid chip for colorectal cancer. Background Technology

[0002] Colorectal cancer, a prevalent malignant tumor worldwide, has long maintained a high incidence and mortality rate, posing a significant public health threat to human health. In the process of basic research and drug development in colorectal cancer, the authenticity and reliability of in vitro models directly determine the translational value of research results. Traditional in vitro cell culture models cannot replicate the complex physiological microenvironment of tumors, leading to significant discrepancies between research conclusions and actual in vivo conditions; animal models, on the other hand, are affected by interspecies biological differences, making it difficult to accurately predict drug responses in human patients.

[0003] Organoid-on-a-chip technology, by combining the core advantages of organoid and microfluidic chip technologies, provides a breakthrough solution for constructing high-fidelity in vitro tumor models. Currently developed organoid chips for colorectal cancer primarily consist of three parts: a porous membrane carrier, a microfluidic control system, and a detection module. The microfluidic control system, with its multi-channel inlet design, enables precise delivery of culture medium, drugs, and growth factors. Combined with a flow control unit to regulate fluid shear force, it simulates the mechanical stimulation generated by intestinal peristalsis. Simultaneously, it can regulate microenvironmental parameters through multi-dimensional external interventions such as temperature, light, and electric fields, simulating the dynamic changes of the tumor microenvironment and meeting the personalized needs of different research scenarios. The detection module can monitor key indicators in the microenvironment in real time, such as pH, oxygen partial pressure, and nutrient concentration, achieving precise monitoring throughout the experimental process and significantly improving the reliability and reproducibility of research results.

[0004] Although such structures can reproduce the characteristics of the tumor microenvironment to a certain extent and be applied to drug activity testing, there are still obvious limitations. They cannot fully simulate the complex interactions between colorectal cancer tissue and multiple systems of the body, such as the immune system and nervous system. This has become a key bottleneck restricting its further promotion and application. Utility Model Content

[0005] The purpose of this invention is to propose an organoid chip for colorectal cancer to solve the problems mentioned in the background art.

[0006] To achieve the above objectives, the technical solution adopted by this utility model is as follows: a colorectal cancer organoid chip, comprising a chip substrate, on which a composite functional layer connected to a microfluidic control system and a detection module is provided; the composite functional layer consists of a support layer, a vascularization layer, a tumor microenvironment simulation layer and an epithelial barrier layer from bottom to top, to respectively simulate different structural levels of colorectal cancer tissue and the surrounding microenvironment, and the layers are connected by a nanofiber bridging structure to realize interlayer material exchange and signal transduction.

[0007] Preferably, the chip substrate is made of glass, which has good optical transparency and stability, making it easy to observe and operate.

[0008] Preferably, the support layer is made of PDMS material and has a reinforcing rib structure inside to improve the overall structural stability without affecting the permeability of materials.

[0009] Preferably, the vascularization layer includes a hydrogel matrix for inoculating vascular endothelial cells and perivascular cells, wherein a vascular network channel is embedded within the hydrogel matrix.

[0010] Preferably, the vascular network channels are distributed in a hierarchical branching pattern, and their surfaces are modified with vascular endothelial growth factor to promote the adhesion and growth of vascular endothelial cells and construct a functional vascular network.

[0011] Preferably, the tumor microenvironment simulation layer includes a hydrogel scaffold, and the hydrogel scaffold has multiple circular organoid culture cavities for inoculating colorectal cancer cells.

[0012] Preferably, the hydrogel scaffold is loaded with fibronectin and laminin.

[0013] Preferably, the cavity wall of the organoid culture chamber is provided with a micro-protrusion structure to enhance cell adhesion and provide a suitable space for organoid growth.

[0014] Preferably, the epithelial barrier layer comprises a nanofiber porous membrane, the surface of which is modified with cell adhesion molecules to mimic the function of the intestinal epithelial barrier.

[0015] Preferably, the nanofiber porous membrane is made of polylactic acid-glycolic acid copolymer.

[0016] Beneficial effects: Compared with existing technologies, this invention restores the hierarchical anatomical features of colorectal cancer tissue through a layered design of a composite functional layer, including a support layer, a vascularization layer, a tumor microenvironment simulation layer, and an epithelial barrier layer. The nanofiber bridging structure between each layer realizes physiological interlayer material exchange and signal transduction, helping to simulate the interaction between the tumor and multiple systems such as the immune system. In conjunction with the microfluidic control system and detection module, the overall design significantly improves the realism of the microenvironment simulation, the reliability of the research results, and the translational value, effectively solving the bottleneck problem of incomplete simulation in existing chips. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the structure of this utility model; Figure 2 for Figure 1 A magnified view of a portion of the image; Figure 3 This is a schematic diagram of the reinforcing rib structure in this utility model; Figure 4 This is a schematic diagram of the vascularization layer in this utility model; Figure 5 This is a schematic diagram of the tumor microenvironment simulation layer in this invention; Figure 6 This is a schematic diagram of the epithelial barrier layer in this utility model.

[0018] Figure labels: 1. Chip substrate; 2. Composite functional layer; 21. Support layer; 211. Reinforcing rib structure; 22. Vascularization layer; 221. Hydrogel matrix; 222. Vascular network channel; 223. Vascular endothelial growth factor; 23. Tumor microenvironment simulation layer; 231. Hydrogel scaffold; 232. Organoid culture cavity; 233. Microprotrusion structure; 24. Epithelial barrier layer; 241. Nanofiber porous membrane; 242. Cell adhesion molecule; 25. Nanofiber bridging structure. Detailed Implementation

[0019] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0020] See attached document Figure 1As shown, this invention proposes an organoid microarray for colorectal cancer, comprising a chip substrate 1. The chip substrate 1 is made of 1.5mm thick glass, possessing good optical transparency and stability, facilitating observation and operation. A composite functional layer 2 is provided on the chip substrate 1, connected to a microfluidic control system and a detection module, to achieve precise simulation and research of the colorectal cancer microenvironment.

[0021] See attached document Figure 2 As shown, the composite functional layer 2 consists of a support layer 21, a vascularization layer 22, a tumor microenvironment simulation layer 23, and an epithelial barrier layer 24 from bottom to top. Each layer simulates different structural levels and surrounding microenvironment of colorectal cancer tissue. The layers are connected by nanofiber bridging structures 25 to achieve interlayer material exchange and signal transduction, which highly restores the hierarchical structure and microenvironment complexity in colorectal cancer tissue and solves the problem of incomplete simulation in existing chips.

[0022] When in use, the chip is pre-treated and sterilized before being layered and inoculated with corresponding cells for culture. By working in conjunction with the microfluidic control system and detection module, it simulates the intestinal fluid environment, restores the in vivo microenvironment, and then conducts experiments such as drug screening. The results are then analyzed by combining real-time monitoring and optical observation.

[0023] See attached document Figure 3 As shown, the support layer 21 is made of PDMS material with a thickness of 400μm and has a reinforcing rib structure 211 inside to improve the overall structural stability without affecting the material permeability.

[0024] See attached document Figure 4 As shown, the vascularized layer 22 has a thickness of 250 μm and includes a hydrogel matrix 221. The hydrogel matrix 221 is used to seed vascular endothelial cells and perivascular cells, and vascular network channels 222 are embedded within the hydrogel matrix 221. The vascular network channels 222 are distributed in a hierarchical branching manner, with the main trunk having a diameter of 300 μm and the branch channels having a diameter of 80 μm. The surface of the vascular network channels 222 is modified with vascular endothelial growth factor 223 to promote the adhesion and growth of vascular endothelial cells and construct a functional vascular network.

[0025] See attached document Figure 5 As shown, the tumor microenvironment simulation layer 23 has a thickness of 350 μm and includes a hydrogel scaffold 231 loaded with fibronectin and laminin. The hydrogel scaffold 231 contains multiple circular organoid culture cavities 232 with a diameter of 800 μm for inoculating colorectal cancer cells. The cavity walls of the organoid culture cavities 232 have micro-protrusion structures 233 with a height of 30 μm to enhance cell adhesion and provide a suitable space for organoid growth.

[0026] See attached document Figure 6As shown, the epithelial barrier layer 24 has a thickness of 80 μm and includes a nanofiber porous membrane 241. The nanofiber porous membrane 241 is made of polylactic acid-glycolic acid copolymer, with a pore diameter of 100 nm and a porosity of 70%. The surface of the nanofiber porous membrane 241 is modified with cell adhesion molecules 242 to simulate the function of the intestinal epithelial barrier.

[0027] The foregoing has shown and described the basic principles, main features, and advantages of this utility model. Those skilled in the art should understand that this utility model is not limited to the above embodiments. The descriptions of the above embodiments and specifications are merely illustrative of the principles of this utility model. Various changes and modifications can be made to this utility model without departing from its spirit and scope, and all such changes and modifications fall within the scope of protection claimed by this utility model. The scope of protection of this utility model is defined by the appended claims and their equivalents.

Claims

1. An organoid chip for colorectal cancer, comprising a chip substrate (1), wherein a composite functional layer (2) connected to a microfluidic control system and a detection module is provided on the chip substrate (1); characterized in that: The composite functional layer (2) consists of a support layer (21), a vascularization layer (22), a tumor microenvironment simulation layer (23), and an epithelial barrier layer (24) from bottom to top, respectively simulating different structural levels and surrounding microenvironment of colorectal cancer tissue. The layers are connected by a nanofiber bridging structure (25) to achieve interlayer material exchange and signal transduction.

2. The colorectal cancer organoid microarray according to claim 1, characterized in that: The support layer (21) is made of PDMS material, and the support layer (21) has a reinforcing rib structure (211) inside.

3. The colorectal cancer organoid microarray according to claim 2, characterized in that: The vascularized layer (22) includes a hydrogel matrix (221) for seeding vascular endothelial cells and perivascular cells, and the hydrogel matrix (221) contains embedded vascular network channels (222).

4. The colorectal cancer organoid microarray according to claim 3, characterized in that: The vascular network channels (222) are distributed in a hierarchical branching pattern, and their surfaces are modified with vascular endothelial growth factor (223).

5. The colorectal cancer organoid microarray according to claim 4, characterized in that: The tumor microenvironment simulation layer (23) includes a hydrogel scaffold (231), and the hydrogel scaffold (231) contains multiple circular organoid culture cavities (232) for inoculating colorectal cancer cells.

6. The colorectal cancer organoid microarray according to claim 5, characterized in that: The hydrogel scaffold (231) is loaded with fibronectin and laminin.

7. The colorectal cancer organoid microarray according to claim 6, characterized in that: The organoid culture cavity (232) has a micro-protrusion structure (233) on its cavity wall.

8. The colorectal cancer organoid microarray according to claim 7, characterized in that: The epithelial barrier layer (24) includes a nanofiber porous membrane (241) with cell adhesion molecules (242) modified on its surface.

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