Multi-tissue co-culture organoid chip

Abstract

The utility model relates to the technical field of biological chips, in particular to a multi-tissue co-culture organoid chip which comprises a chip main body, and a plurality of culture hole groups are formed in the chip main body; the culture hole group comprises a first cell culture hole and at least one second cell culture hole communicated with the first cell culture hole, and a porous membrane for cells to pass through is arranged at the communicated part of the first cell culture hole and each second cell culture hole; the mode that the two cell culture holes are communicated and separated through the porous membrane is adopted, organoid co-culture is achieved, meanwhile, the pathological and physiological dynamic microenvironment of tumor tissue can be simulated, the operation process of medicine screening is simplified, and the medicine screening efficiency is greatly improved.

Description

Technical field:

[0001] The utility model relates to the technical field of biochips, and in particular to a multi-tissue co-culture organoid chip. Background technology:

[0002] Organoids are miniature three-dimensional structures that are cultured in vitro and can self-assemble. This structure is highly similar to the structure of tissues and organs in the body, contains various cell types, and can self-renew, simulating some of the functions of tissues and organs in the body. Various studies have shown that organoids can reproduce the structure and physiological functions of human organs in detail, and can be used to elucidate pathology and drug development. In order to comply with the trend of shortening clinical testing cycles and replacing animal experiments, research on organoids is prevalent. Research based on biological research combined with engineering technology called organ-on-a-chip for cultivation and analysis is prevalent, collectively referred to as biomimetic systems (microphysiological systems: MPS). As a new biological model, organoids can be used for drug screening and disease modeling due to their unique simulation properties. They have great potential in disease mechanism research, drug screening, regenerative medicine, biomaterial evaluation, etc.

[0003] In recent years, rapid, efficient high-throughput screening technologies have been widely developed and applied, becoming a key technology for drug screening. Currently, biochips used for organoid culture consist of a chip body with multiple microwells within which organoids are cultured. However, these biochips have independent microwells, providing only a culture function and failing to simulate the dynamic microenvironment of tumor tissue pathophysiology. Drug screening also requires transferring the organoids from the chip to a specialized co-culture chip, which is cumbersome and inefficient. Utility model content:

[0004] The purpose of this utility model is to address the deficiencies in the existing technology and provide a multi-tissue co-culture organoid chip that can achieve organoid co-culture and simulate the dynamic microenvironment of tumor tissue pathophysiology, simplify the operation process of drug screening, and greatly improve drug screening efficiency.

[0005] In order to achieve the above-mentioned purpose, the technical solution adopted by the present invention is: a multi-tissue co-culture organoid chip, comprising a chip body, on which a plurality of culture well groups are opened; the culture well groups include a first cell culture well and at least one second cell culture well connected to the first cell culture well, and the connection between the first cell culture well and each second cell culture well is provided with a porous membrane for cells to pass through.

[0006] As a further preferred embodiment, the culture well group is a long strip-shaped slot, and the porous membrane is arranged in the long strip-shaped slot to separate the long strip-shaped slot into a first cell culture well and a second cell culture well.

[0007] As a further preferred embodiment, two porous membranes are provided in the culture well group, and the two porous membranes separate the long slots into a first cell culture well in the middle and two second cell culture wells at both ends.

[0008] As a further preferred embodiment, the plurality of culture well groups are arranged and distributed in a matrix manner.

[0009] As a further preferred embodiment, the corresponding porous membranes in all culture well groups in the same row are interconnected to form an integrated structure.

[0010] As a further preferred embodiment, in the culture well group, the side of the bottom of the second cell culture well facing away from the first cell culture well is inclined upward to form an inclined surface facing the first cell culture well.

[0011] As a further preferred solution, a water tank surrounding all the culture well groups is provided on the chip body.

[0012] As a further preferred solution, a card holder is provided on the periphery of the chip body.

[0013] As a further preferred embodiment, the material of the chip body is selected from at least one of the following: polypropylene resin, polyethylene resin, ethylene-propylene copolymer, polystyrene resin, acrylonitrile-butadiene-styrene resin, polycarbonate resin, polyethylene terephthalate resin, polymethyl methacrylate resin, vinyl chloride resin, polybutylene terephthalate resin, polyarylate resin, polysulfone resin, polyethersulfone resin, polyetheretherketone resin, polyetherimide resin, polytetrafluoroethylene, polymethylpentene resin, and polyacrylonitrile resin.

[0014] As a further preferred solution, a plurality of micropores are distributed on the porous membrane, and the pore diameter of the micropores is 8-12 μm.

[0015] The beneficial effects of the present invention are as follows: the present invention provides a multi-tissue co-culture organoid chip, comprising a chip body, on which a plurality of culture well groups are opened; the culture well groups include a first cell culture well and at least one second cell culture well connected to the first cell culture well, and the connection between the first cell culture well and each second cell culture well is provided with a porous membrane for cells to pass through; when the present invention is in use, normal tissue cells are placed in the first cell culture well for culture, and tumor cells are placed in the second cell culture well for culture, when the normal tissue cells in the first cell culture well are attractive to the tumor cells in the second cell culture well, the tumor cells will pass through the porous membrane into the first cell culture well, and by observing and analyzing the cell conditions in the first cell culture well and the second cell culture well, relevant research such as tissue development, drug safety evaluation, drug penetration and drug screening can be carried out; the present invention adopts a method of connecting the two cell culture wells and separating them with a porous membrane to achieve organoid co-culture while simulating the dynamic microenvironment of tumor tissue pathophysiology, simplifying the operation process of drug screening, and greatly improving the efficiency of drug screening. Description of the drawings:

[0016] Figure 1 It is a structural diagram of the present utility model.

[0017] Figure 2 It is a cross-sectional schematic diagram of the culture well group of the present invention.

[0018] Explanation of the reference numerals: chip body 1 , culture well group 2 , first cell culture well 3 , second cell culture well 4 , porous membrane 5 , inclined surface 41 , water tank 6 , tray 7 , first cell culture well bottom surface 31 , second cell culture well bottom surface 42 . Specific implementation method:

[0019] The present invention will be further described below with reference to the accompanying drawings. Figure 1As shown, the utility model includes a chip body 1, and a plurality of culture well groups 2 are provided on the chip body 1; the culture well group 2 includes a first cell culture well 3, at least one second cell culture well 4 connected to the first cell culture well 3, and the connection between the first cell culture well 3 and each second cell culture well 4 is provided with a porous membrane 5 for cells to pass through. Several micropores are evenly distributed on the porous membrane 5, the pore size of the micropores is preferably 8-12 μm, the thickness of the porous membrane 5 is preferably 8-12 μm, and the porous membrane 5 is preferably a transparent polycarbonate membrane. The first cell culture well 3 and the second cell culture well 4 are connected by the porous membrane 5 to realize the exchange of substances in the culture medium and cell migration. The porous membrane 5 can not only realize the permeation and exchange of nutrients, inorganic salt ions and other substances between the two cell culture wells, but also realize the transmembrane migration of cells of different sizes below 20 μm between the two cell culture wells, thereby allowing the interaction between cells and simulates the microenvironment in the body.

[0020] The culture well group 2 is a rectangular long slot, and the porous membrane 5 is set in the long slot and separates the long slot to form a first cell culture well 3 and a second cell culture well 4. This culture well group 2 has a simpler structure and is easier to manufacture.

[0021] In the present embodiment, the group culture well group 2 has 24 groups, and the 24 groups of culture well groups 2 are arranged in a matrix of 12×2, and the culture well group 2 is arranged in the longitudinal direction. Two porous membranes 5 are provided in the culture well group 2, and the two porous membranes 5 are distributed in parallel and spaced apart in the longitudinal direction, and the long strip slots are separated to form a first cell culture well 3 located in the middle position and two second cell culture wells 4 located at both ends. The corresponding porous membranes 5 in all the culture well groups 2 located in the same row are interconnected into an integrated structure, and processing and molding are simpler, reducing production costs and submitting efficiency. The size and spacing of each first and second cell culture well 4 match the 384-well plate, and can be adapted to various detection instruments using 384-well plates, with strong versatility. Of course, the size and spacing of each first and second cell culture well 4 can also match the 96-well plate and the 1536-well plate.

[0022] In the culture well group 2, the side of the bottom of the second cell culture well 4 facing away from the first cell culture well 3 is tilted upward to form an inclined surface 41 facing the first cell culture well 3. The inclined surface 41 makes the bottom of the second cell culture well 4 have a liquid gathering effect, which is convenient for the pipette to aspirate liquid, and greatly reduces the area of ​​the bottom surface 42 of the second cell culture well, reducing residue and making it easier to remove the residue; on the other hand, the inclined surface 41 of the second cell culture well 4 can make the cells automatically gather at the bottom surface 42 of the second cell culture well. To observe the cells, it is only necessary to observe the area of ​​the bottom surface 42 of the second cell culture well without observing the area of ​​the inclined surface 41, which greatly reduces the area of ​​the observation area and facilitates positioning observation. In addition, the bottom surface 42 of the second cell culture well is connected to the bottom surface 31 of the first cell culture well, so that the bottom surface 42 of the second cell culture well and the bottom surface 31 of the first cell culture well can be included in the observation field of view at the same time, and there is no need to shift the field of view to observe the first cell culture well 3 and the second cell culture well 4 separately, which greatly improves the efficiency of detection.

[0023] The chip body 1 is provided with a water tank 6 surrounding all culture well groups 2. This arrangement of water tank 621 surrounding the culture well groups 2 maintains a moist environment during cell culture and significantly reduces optical errors in the outer ring of microwells during detection. A retainer 7 is provided around the perimeter of the chip body 1 for convenient attachment to a support. To enhance or reduce cell adhesion to the inner walls of the culture wells, the inner walls of the first and second cell culture wells 3 and 4 can be coated with a hydrophilic or hydrophobic coating.

[0024] The material of the chip body 1 is selected from at least one of the following: polypropylene resin, polyethylene resin, ethylene-propylene copolymer, polystyrene resin, acrylonitrile-butadiene-styrene resin, polycarbonate resin, polyethylene terephthalate resin, polymethyl methacrylate resin, vinyl chloride resin, polybutylene terephthalate resin, polyarylate resin, polysulfone resin, polyethersulfone resin, polyetheretherketone resin, polyetherimide resin, polytetrafluoroethylene, polymethylpentene resin, and polyacrylonitrile resin. Polystyrene resin is preferred in view of the moldability and sterilizability required for the culture container.

[0025] When the present invention is in use, normal tissue cells are placed in the first cell culture well 3 for culture, and tumor cells are placed in the second cell culture well 4 for culture. When the normal tissue cells in the first cell culture well 3 are attractive to the tumor cells in the second cell culture well 4, the tumor cells will pass through the porous membrane 5 and enter the first cell culture well 3. By observing and analyzing the cell conditions in the first cell culture well 3 and the second cell culture well 4, relevant research such as tissue development, drug safety evaluation, drug penetration and drug screening can be carried out; the present invention adopts the method of connecting the two cell culture wells and separating them with the porous membrane 5 to achieve organoid co-culture while simulating the dynamic microenvironment of tumor tissue pathophysiology, simplifying the operation process of drug screening, and greatly improving the efficiency of drug screening.

[0026] Of course, the above is only a preferred embodiment of the present invention, so all equivalent changes or modifications made according to the structure, features and principles described in the scope of the present invention patent application are included in the scope of the present invention patent application.

Claims

1. A multi-tissue co-culture organoid chip, comprising a chip body (1), characterized in that: The chip body (1) is provided with a plurality of culture well groups (2); the culture well groups (2) include a first cell culture well (3), at least one second cell culture well (4) connected to the first cell culture well (3), and a porous membrane (5) through which cells can pass is provided at the connection points between the first cell culture well (3) and each second cell culture well (4).

2. The multi-tissue co-culture organoid chip according to claim 1, characterized in that: The culture well group (2) is a long strip-shaped slot, and the porous membrane (5) is arranged in the long strip-shaped slot and separates the long strip-shaped slot to form a first cell culture well (3) and a second cell culture well (4).

3. The multi-tissue co-culture organoid chip according to claim 2, characterized in that: Two porous membranes (5) are provided in the culture well group (2), and the two porous membranes (5) separate the long strip slots to form a first cell culture well (3) located in the middle and two second cell culture wells (4) located at both ends.

4. The multi-tissue co-culture organoid chip according to claim 2 or 3, characterized in that: The plurality of culture well groups (2) are arranged and distributed in a matrix manner.

5. The multi-tissue co-culture organoid chip according to claim 4, characterized in that: The corresponding porous membranes (5) in all the culture well groups (2) located in the same row are connected to each other to form an integrated structure.

6. The multi-tissue co-culture organoid chip according to claim 1, characterized in that: In the culture well group (2), the bottom of the second cell culture well (4) is tilted upward on the side facing away from the first cell culture well (3) to form an inclined surface (41) facing the first cell culture well (3).

7. The multi-tissue co-culture organoid chip according to claim 1, characterized in that: The chip body (1) is provided with a water tank (6) surrounding all the culture well groups (2).

8. The multi-tissue co-culture organoid chip according to claim 1, characterized in that: A card holder (7) is provided on the periphery of the chip body (1).

9. The multi-tissue co-culture organoid chip according to claim 1, characterized in that: The material of the chip body (1) is selected from at least one of the following: polypropylene resin, polyethylene resin, ethylene-propylene copolymer, polystyrene resin, acrylonitrile-butadiene-styrene resin, polycarbonate resin, polyethylene terephthalate resin, polymethyl methacrylate resin, vinyl chloride resin, polybutylene terephthalate resin, polyarylate resin, polysulfone resin, polyethersulfone resin, polyetheretherketone resin, polyetherimide resin, polytetrafluoroethylene, polymethylpentene resin, and polyacrylonitrile resin.

10. The multi-tissue co-culture organoid chip according to claim 1, characterized in that: The porous membrane (5) is provided with a plurality of micropores, each of which has a pore diameter of 8-12 μm.

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