High-throughput organoid culture liquid core device

Abstract

The invention discloses a high-flux organoid culture liquid core device which comprises an upper cover module, a transwell module and a porous culture plate, more than one culture hole is formed in the porous culture plate, a liquid inlet hole and a liquid outlet hole are formed in the bottom of each culture hole, at least one culture liquid circulation channel is formed in the porous culture plate, and the culture liquid circulation channels are communicated with the culture holes. The culture solution circulation channel comprises an injection port, an input channel, a discharge channel and a discharge port, the injection port is communicated with the input channel, and the discharge channel is communicated with the discharge port; the porous culture plate comprises a plurality of culture holes and a plurality of culture solution circulation channels, each culture solution circulation channel is communicated with an injection port and a discharge port of at least one culture hole, the transwell module comprises more than one transwell chamber, the transwell chambers are matched with the culture holes and placed on the culture holes, and the upper cover module is used for sealing and covering the porous culture plate. According to the liquid core device, a traditional culture plate and a micro-fluidic technology are combined, high-throughput culture of the organoid is achieved, and the homogeneity of the organoid culture tissue is improved due to the fact that a parallel multi-group organoid culture mode is adopted.

Description

Technical Field

[0001] The present invention belongs to the technical field of organoid culture, and in particular relates to a high-throughput organoid culture liquid core device. Background Art

[0002] Organoids are in vitro organ models, primarily cell-tissue complexes formed through stem cell self-organization under three-dimensional (3D) culture conditions. Organoids typically share some key structural and functional characteristics of human tissues or organs. They offer advantages such as cryopreservation and thawing, long-term culture, widespread availability, high success rates, short modeling times, and relatively simple culture systems. They have shown promising applications in areas such as tissue and organ development, disease simulation, drug screening, and regenerative medicine.

[0003] At present, a series of organoid models for various human organs (such as the brain, liver, kidney, and intestine) have been established and have begun to be used in research in the fields of biology, medicine, and pharmacy. In the study of various clinical diseases, organoid models have always been an important tool for life science research. For a long time, two-dimensional cell models and animal models have been the main models for simulating human development and disease. Among them, animal models can provide a certain in vivo physiological environment compared to cell lines, but animal models are very complex and require a lot of manpower, material resources, and financial resources. At the same time, due to differences between species. The above models have certain shortcomings: they are manifested in low degree of biomimetic, difficult to characterize and observe, species differences, and ethical issues. It is difficult to achieve high-throughput screening analysis in the field of drug or compound screening analysis.

[0004] Organoids, a novel experimental system distinct from 2D cell lines and animal models, can partially mimic the 3D structure of tissues in vitro while also offering the advantages of cell line passage and high throughput. Organoid culture technology not only alters cells at the cellular level but also restores the structural characteristics of tissues and organs.

[0005] The current method of culturing organoids is mainly static culture in well plates. The operator cuts up the primary sample, then coats it in matrix gel at low temperature, and then adds culture medium and places it in an incubator for culture. The medium is changed three times a week, and the cells are passaged every 7-10 days. This culture model has many problems: First, the growth rate of organoids is slow and the culture cycle is long. During the operation, the culture chamber needs to be opened regularly for manual liquid change, which has a high risk of contamination. When the number of culture samples increases, the cost of manpower, equipment, space, etc. will increase significantly. Second, static culture in the culture plate will lead to insufficient exchange between organoids and culture medium, especially the internal nutrients of 3D cell tissues cannot be absorbed, which will ultimately limit the growth of organoids and limit the growth degree of organoids and their similarity to human organs. Summary of the Invention

[0006] Based on the above-mentioned existing technology, the present invention provides a high-throughput organoid culture liquid core device, which combines traditional culture plates and microfluidic technology. It not only realizes high-throughput organoid culture, but also improves the uniformity of organoid culture tissue due to the use of a parallel multi-group organoid culture mode.

[0007] The technical solution adopted to achieve the above-mentioned purpose of the present invention is: a high-throughput organoid culture liquid core device, including an upper cover module, a transwell module and a multi-porous culture plate, wherein the multi-porous culture plate is provided with more than one culture well, and the bottom of the culture well is respectively provided with a liquid inlet and a liquid outlet, and the multi-porous culture plate is provided with at least one culture liquid flow path, the culture liquid flow path includes an injection port, an input channel, a discharge channel and a discharge port, the injection port and the discharge port are respectively arranged on the side wall of the multi-porous culture plate, the input channel and the discharge channel are arranged on the bottom of the multi-porous culture plate, the input channel and the discharge channel are isolated from each other, the injection port is connected to the input channel, and the discharge channel is connected to the discharge port;

[0008] In each culture fluid flow path, each culture fluid flow path is connected to at least one culture well, and the input channel is connected to the injection port of at least one culture well, and the discharge channel is connected to the discharge port of at least one culture well;

[0009] The transwell module includes one or more transwell chambers, which are adapted to the culture wells and placed on the culture wells. The upper cover module covers the multi-well culture plate and seals the transwell chambers placed on the multi-well culture plate.

[0010] When there are multiple culture fluid flow paths, any two culture fluid flow paths are isolated from each other.

[0011] The culture wells are distributed in a matrix, the number of culture fluid flow passages is the same as the number of rows of culture wells, the injection port and the discharge port of each culture fluid flow passage are respectively located on both sides of the culture wells in the corresponding row, the input channel and the discharge channel of each culture fluid flow passage are respectively located on both sides of the bottom of the culture wells in the corresponding row, the input channel of each culture fluid flow passage is connected to the liquid inlet of each culture well in the corresponding row, and the discharge channel of each culture fluid flow passage is connected to the liquid outlet of each culture well in the corresponding row.

[0012] In each culture fluid flow path, one end of the input channel is sealed, and the other end of the input channel is connected to the injection port; one end of the discharge channel is sealed, and the other end of the discharge channel is connected to the discharge port.

[0013] The multi-porous culture plate includes a multi-porous culture plate body and a channel sealing plate. More than one group of culture fluid circulation grooves are opened on the bottom of the multi-porous culture plate body. Each group of culture fluid circulation grooves includes an input groove and a discharge groove. The channel sealing plate seals the bottom of the multi-porous culture plate body. The space formed by the input groove of each group of culture fluid circulation grooves and the channel sealing plate constitutes the input channel of the corresponding culture fluid circulation path. The space formed by the discharge groove of each group of culture fluid circulation grooves and the channel sealing plate constitutes the discharge channel of the corresponding culture fluid circulation path.

[0014] The multi-well culture plate body is provided with coding marks distributed in a matrix, and each coding mark is located outside the bottom of the corresponding culture well.

[0015] The multi-porous culture plate is square or round. If the multi-porous culture plate is square, the injection port and the discharge port of each culture fluid flow path are respectively arranged on the two side walls in the width direction of the multi-porous culture plate, and the straight line of each row of culture holes is parallel to the length direction of the multi-porous culture plate.

[0016] The bottom of the transwell chamber is sealed, and a plurality of semipermeable membrane windows uniformly distributed along the circumferential direction are provided on the side wall of the transwell chamber.

[0017] The transwell chamber comprises a transwell chamber body, a semipermeable membrane and a fixed sleeve. The bottom of the transwell chamber body is sealed, the bottom of the transwell chamber body is disc-shaped, the side wall of the transwell chamber body is provided with a plurality of first through grooves evenly distributed along the circumferential direction, the lower part of the side wall of the transwell chamber body is annular, the first through grooves evenly divide the lower part of the side wall of the transwell chamber body into mutually isolated inner pressure plates, the outer wall of the bottom of the transwell chamber body is connected to the inner wall of each inner pressure plate, the semipermeable membrane is annular, the fixed sleeve is annular, and the fixed sleeve is provided with a plurality of through grooves evenly distributed along the circumferential direction. The second through grooves are evenly distributed in the direction, and the second through grooves cooperate with the corresponding first through grooves. A snap ring is provided at the bottom of the inner wall of the fixed sleeve, and the outer wall of the snap ring is fitted with the bottom of the inner wall of the fixed sleeve. A plurality of protrusions evenly distributed along the circumferential direction are provided on the top of the snap ring. A snap groove for fixing the semipermeable membrane is respectively provided between the two ends of each protrusion and the inner wall of the fixed sleeve. The semipermeable membrane is fitted to the inner wall of the fixed sleeve, and the lower end of the semipermeable membrane is inserted into each snap groove. The lower part of the side wall of the transwell chamber body is embedded in the semipermeable membrane, the bottom of each inner pressure plate is located on the top of the snap ring, the bottom of the transwell chamber body is embedded in the snap ring, and the bottom of the transwell chamber body is interference fit with the snap ring.

[0018] The first through groove extends to the top of the side wall of the transwell chamber body. The side wall of the transwell chamber body is composed of multiple internal pressure plates. Arc-shaped connecting strips are provided between adjacent internal pressure plates. Each connecting strip is connected to the middle part of two adjacent internal pressure plates. The top of the fixed sleeve is connected to each connecting strip by a snap buckle.

[0019] The upper cover module includes an upper cover body and a sealing ring. The upper cover body covers the top of the multi-porous culture plate. The bottom of the outer wall of the upper cover body covering the sealing part is provided with a sealing groove that cooperates with the sealing plate, and the sealing ring is sleeved in the sealing groove.

[0020] The outer side wall of the upper cover body is provided with an anti-slip tooth structure, and the corresponding position of the outer side wall of the multi-porous culture plate is provided with an anti-slip tooth structure.

[0021] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0022] 1. Compared with traditional organoid culture models, the present invention can not only simultaneously culture organoids through transwell chambers in multiple groups of culture wells, achieving high-throughput culture of organoids, but also construct a parallel multi-group organoid culture system to improve the uniformity of organoid culture tissues. It can also conduct control experiments on organoid culture. Since it is a parallel multi-group organoid culture system, the accuracy and reliability of the experiment are improved.

[0023] 2. The present invention connects multiple groups of culture wells in parallel to achieve high-throughput continuous perfusion of cell culture fluid in organoid culture, increasing the high-throughput of organoid model construction and improving the efficiency and accuracy of drug or chemical screening and analysis. It can also observe and analyze the differential responses of organoids from different patient sources to drugs, thereby improving the accuracy of comparative experiments on organoids from different patient sources.

[0024] 3. The liquid core device of the present invention adopts modular composition, which is convenient to install and disassemble, and can be flexibly and quickly replaced and assembled.

[0025] 4. The size of the liquid core device and the structural design of the culture holes and perfusion channels of the present invention can better meet the perfusion of high-throughput culture fluid and adapt to various characterization equipment, thereby achieving high-throughput characterization.

[0026] 5. The upper cover body of the liquid core device of the present invention and the outer side wall of the cell culture plate body are both provided with tooth-like structures, which not only facilitates the experimenter's handling operation but also enhances the strength of the multi-well culture plate.

[0027] 6. The present invention has coding marks designed on the culture plate body at the periphery of the culture wells to distinguish and identify the culture wells, thereby reducing the risk of misoperation.

[0028] 7. The transwell chamber of the present invention is provided with multiple semipermeable membrane windows evenly distributed along the circumferential direction on the side wall, which changes the traditional transwell chamber mode of setting the semipermeable membrane at the bottom. This allows the transwell chamber with this special structure to better cooperate with the culture well plate with continuous dynamic perfusion channels, thereby more realistically simulating the bionic environment of organoid culture. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Schematic diagram of the structure of the high-throughput organoid culture liquid core device.

[0030] Figure 2 Exploded view of the high-throughput organoid culture liquid core device.

[0031] Figure 3 A top view of the high-throughput organoid culture liquid core device.

[0032] Figure 4 for Figure 3 Cross-sectional view along the AA axis.

[0033] Figure 5 for Figure 4 Cross-sectional view along CC direction.

[0034] Figure 6 for Figure 3 Cross-sectional view along the BB direction.

[0035] Figure 7 for Figure 3 Cross-sectional view along the DD direction.

[0036] Figure 8 Schematic diagram of the structure of the transwell chamber.

[0037] Figure 9 Exploded view of the transwell chamber.

[0038] Figure 10 It is a structural diagram of the fixed sleeve.

[0039] Among them, 1-upper cover module: 101-upper cover body, 102-sealing ring; 2-transwell module: 201-transwell chamber: 201-transwell chamber body, 202-translucent, 203-fixing sleeve, 204-first through groove, 205-inner pressure plate, 206-connecting strip, 207-buckle, 208-second through groove, 209-clamping ring, 210-protrusion, 211-slot; 3-multi-porous culture plate body: 301-multi-porous culture plate body, 302-channel sealing plate; 4-culture well: 401-liquid inlet, 402-liquid inlet; 5-culture fluid flow path: 501-injection port, 502-input channel, 503-discharge channel, 504-discharge port; 6-coding mark; 7-tooth structure. DETAILED DESCRIPTION

[0040] The present invention will be described in detail below with reference to the accompanying drawings.

[0041] The high-throughput organoid culture liquid core device provided in this embodiment is as follows Figure 1 and Figure 2 As shown, the liquid core device is square in shape. Of course, the liquid core device can be of other shapes, such as round. In this embodiment, the liquid core device is designed to be square. The core device is compatible with various characterization equipment and can be easily characterized. The liquid core device includes a cover module 1, a transwell module 2, and a multi-well culture plate 3.

[0042] Transwell module 2 includes 12 transwell chambers, each of which can culture and form an organoid model.

[0043] like Figure 7-10 As shown, the transwell chamber includes a transwell chamber body 201, a semipermeable membrane 202 and a fixed sleeve 203, and the semipermeable membrane 202 and the fixed sleeve 203 are both annular. The transwell chamber body 201 is in an inverted cone shape as a whole, the bottom of the transwell chamber body 201 is sealed, the bottom of the transwell chamber body 201 is disc-shaped, and the lower part of the side wall of the transwell chamber body 201 is annular. The side wall of the transwell chamber body 201 is provided with three first through grooves 204 evenly distributed along the circumferential direction. The first through grooves 204 evenly divide the side wall of the transwell chamber body into mutually isolated inner pressure plates 205. The outer wall of the bottom of the transwell chamber body 201 is connected to the inner wall of each inner pressure plate 205. An arc-shaped connecting strip 206 is provided between adjacent inner pressure plates 205, and each connecting strip 205 is connected to the middle part of two adjacent inner pressure plates 205.

[0044] The fixing sleeve is provided with three second through-grooves 208 evenly distributed along the circumference, which mate with corresponding first through-grooves 204. A snap ring 209 is provided at the bottom of the inner wall of the fixing sleeve. The outer wall of snap ring 209 mates with the bottom of the inner wall of the fixing sleeve 203, forming a single-piece structure. The top of snap ring 209 is provided with three protrusions 210 evenly distributed along the circumference. A snap groove 211 for securing the semipermeable membrane is provided between the ends of each protrusion 210 and the inner wall of the fixing sleeve 203. The semipermeable membrane 202 is attached to the inner wall of the fixing sleeve 203, with the lower end of the semipermeable membrane 202 inserted into each snap groove 211. The lower portion of the sidewall of the transwell chamber body 201 is embedded in the semipermeable membrane 202, and the bottom of each inner pressure plate 205 is located on the top of the snap ring 209. The bottom of the transwell chamber body 201 is embedded in the snap ring 209, creating an interference fit between the bottom of the transwell chamber body 201 and the snap ring 209. The top surface of the bottom of the transwell chamber body 201 is flush with the top surface of each protrusion 210. Each connecting strip 206 is provided with a snap 207, and the top of the fixing sleeve 203 is connected to each connecting strip 206 via the snap 207.

[0045] Each first through-groove 204, the corresponding second through-groove 208, and the semipermeable membrane 202 form a semipermeable membrane window, allowing the inflow and outflow of culture fluid. The provision of these side semipermeable membrane windows, combined with the multi-well culture plate 3 having continuous dynamic perfusion channels, enables continuous dynamic perfusion of culture fluid during organoid culture, thereby simulating a biomimetic organoid culture environment.

[0046] The multi-well plate 3 can be custom-made by 3D printing or injection molding using transparent resin materials. Transparent resin materials include biocompatible transparent materials such as PS (polystyrene), PMMA (polymethyl methacrylate), PC (polycarbonate), PET (polyethylene terephthalate), PP (polypropylene), and COC / COP (cyclic olefin polymer). The multi-well plate 3 is square in shape and includes a multi-well plate body 301 and a channel sealing plate 302. Both the multi-well plate body 301 and the channel sealing plate 302 are square in shape. Of course, the multi-well plate 1 can also have other shapes, such as a circle.

[0047] The multi-porous culture plate body 301 is provided with culture wells 4 distributed in a matrix. Of course, the culture wells 4 can also be arranged in a circular array or other ways according to actual needs. In this embodiment, the culture wells 4 are distributed in 3 rows × 4 columns, and the number of rows and columns of the culture wells 4 can also be designed according to actual needs to meet actual needs. The straight line in which each row of culture wells 4 is distributed is parallel to the length direction of the multi-porous culture plate 3, and the straight line in which each column of culture wells 4 is located is parallel to the width direction of the multi-porous culture plate 3. The shape of the culture well 4 can be designed according to actual conditions. In this embodiment, the culture well 4 is a circular hole. The size and structure of the culture well 4 are compatible with the transwell chamber 201, and each transwell chamber 201 is placed on the corresponding culture well 4. The transwell chamber is separable from the culture well, which makes it convenient to take out the transwell chamber for characterization, further improving the convenience and accuracy of characterization. The liquid inlet 401 and the liquid outlet 402 are symmetrically provided at the bottom of the culture well 4, and the culture fluid is perfused in the transwell chamber 201 through the liquid inlet 401 and the liquid outlet 402. As Figure 3-7 As shown, the multi-well plate 3 is provided with three parallel culture fluid flow paths 5, each comprising an inlet 501, an input channel 502, an outlet channel 503, and an outlet 504. The three inlets 501 are arranged side by side on one widthwise sidewall of the multi-well plate body 301, while the three outlets 504 are arranged side by side on the other widthwise sidewall of the multi-well plate body 301. The bottom of the multi-well plate body 301 is provided with three parallel sets of culture fluid flow channels, each set comprising an input channel and an outlet channel. A channel sealing plate 302 seals the bottom of the multi-well plate body 301. The space enclosed by the input channel of each set of culture fluid flow channels and the channel sealing plate constitutes the input channel 502 of the corresponding culture fluid flow path, while the space enclosed by the outlet channel of each set of culture fluid flow channels and the channel sealing plate constitutes the outlet channel 503 of the corresponding culture fluid flow path. In each culture fluid flow path, one end of the input channel 502 is sealed, and the other end of the input channel 502 is connected to the injection port 501 ; one end of the discharge channel 503 is sealed, and the other end of the discharge channel 502 is connected to the discharge port 504 .

[0048] like Figure 3-7As shown, the input channel 502 and the discharge channel 503 of each culture fluid circulation path are parallel to the straight line distributed in each row of culture wells 4, and the input channel 502 and the discharge channel 503 of each culture fluid circulation path are respectively located on both sides of the bottom of the corresponding row of culture wells 4. The input channel 502 of each culture fluid circulation path is connected to the liquid inlet 401 of each culture well 4 in the corresponding row, and the discharge channel 503 of each culture fluid circulation path is connected to the liquid outlet 402 of each culture well 4 in the corresponding row. Through the coordination of the liquid inlet 401 and the input channel 502 at the bottom of the culture well 4, and the coordination of the liquid outlet 402 and the discharge channel 503 at the bottom of the culture well 4, a cell culture fluid perfusion channel is constructed, so that the culture wells 4 on the multi-well culture plate 3 are connected in parallel, completely replacing the culture method in which traditional well plates cannot achieve liquid communication, making the cultured organoid models highly uniform, and greatly reducing the experimental errors caused by differences in organoid models.

[0049] The multi-well culture plate body 301 is provided with coding marks 6 distributed in a matrix. Each coding mark 6 is located outside the bottom of the corresponding culture well 4. The culture wells 4 are distinguished and identified by coding, reducing the risk of misoperation.

[0050] like Figure 1-2 As shown, the upper cover module 1 is square in shape and is made of a transparent biocompatible material. The upper cover module 1 includes an upper cover body 101 and a sealing ring 102. Both the upper cover body 101 and the sealing ring 102 are square in shape. The upper cover body 101 covers the top of the multi-well culture plate body 301 and seals the openings of each transwell chamber. The outer wall bottom of the upper cover body 101 covering the sealing portion is provided with a sealing groove that cooperates with the sealing ring 102. The sealing ring 102 is mounted in the sealing groove, and the connection between the upper cover body and the multi-well culture plate body is sealed by the sealing ring.

[0051] like Figure 1-2 As shown, tooth structures 7 are symmetrically provided at the middle parts of the outer walls of the upper cover body 101 in both length directions, and tooth structures 7 are symmetrically provided at the middle parts of the outer walls of the multi-porous culture plate body 301 in both length directions. The tooth structures 7 are provided to prevent slipping, making it convenient for the experimenters to take the entire core device. At the same time, the tooth structures 7 will reduce the deformation of the multi-porous culture plate body 301 to a certain extent.

[0052] The method for using the high-throughput organoid culture liquid core device is as follows:

[0053] S1. Place a single transwell chamber in culture well 4 of a multi-well culture plate.

[0054] S2. After placement is completed, the multi-well culture plate 3 and the upper cover module 1 are sterilized by ultraviolet irradiation or ethylene oxide sterilization;

[0055] S3. Mincing, digesting, filtering, and centrifuging the sample tissue to obtain an organoid suspension corresponding to the sample tissue, uniformly mixing the organoid suspension with a matrix gel, such as Matrigel, in a certain proportion, and then seeding the organoid-matrigel mixture in a transwell chamber 201. The multi-well culture plate is then placed in a CO2 cell culture incubator to solidify the organoid-matrigel mixture.

[0056] S4. After the organoid-matrigel mixture solidifies, the culture medium is pumped into the input injection channel 502 through the injection port 501, and the culture medium enters each transwell chamber 201. When the culture medium flows out of the discharge port 504, the upper cover module 1 is placed on the multi-porous culture plate 3 to seal the transwell chamber 201, and then the organoid model is dynamically cultured.

Claims

1. A high-throughput organoid culture fluid core device, characterized by: The multi-porous culture plate comprises an upper cover module, a transwell module and a multi-porous culture plate, wherein the multi-porous culture plate is provided with more than one culture well, the bottom of each culture well is respectively provided with a liquid inlet and a liquid outlet, the multi-porous culture plate is provided with at least one culture solution flow path, the culture solution flow path comprises an injection port, an input channel, a discharge channel and a discharge port, the injection port and the discharge port are respectively provided on the side wall of the multi-porous culture plate, the input channel and the discharge channel are provided on the bottom of the multi-porous culture plate, the input channel and the discharge channel are isolated from each other, the injection port is connected to the input channel, and the discharge channel is connected to the discharge port; In each culture fluid flow path, each culture fluid flow path is connected to at least one culture well, and the input channel is connected to the injection port of at least one culture well, and the discharge channel is connected to the discharge port of at least one culture well; The transwell module includes one or more transwell chambers, which are adapted to the culture wells and placed on the culture wells. The upper cover module covers the multi-well culture plate and seals the transwell chambers placed on the multi-well culture plate.

2. The high-throughput organoid culture liquid core device according to claim 1, characterized in that: When there are multiple culture fluid flow paths, any two culture fluid flow paths are isolated from each other.

3. The high-throughput organoid culture liquid core device according to claim 2, characterized in that: The culture wells are distributed in a matrix, the number of culture fluid flow passages is the same as the number of rows of culture wells, the injection port and the discharge port of each culture fluid flow passage are respectively located on both sides of the culture wells in the corresponding row, the input channel and the discharge channel of each culture fluid flow passage are respectively located on both sides of the bottom of the culture wells in the corresponding row, the input channel of each culture fluid flow passage is connected to the liquid inlet of each culture well in the corresponding row, and the discharge channel of each culture fluid flow passage is connected to the liquid outlet of each culture well in the corresponding row.

4. The high-throughput organoid culture liquid core device according to claim 3, characterized in that: In each culture fluid flow path, one end of the input channel is sealed, and the other end of the input channel is connected to the injection port; one end of the discharge channel is sealed, and the other end of the discharge channel is connected to the discharge port.

5. The high-throughput organoid culture liquid core device according to claim 3, characterized in that: The multi-porous culture plate includes a multi-porous culture plate body and a channel sealing plate. More than one group of culture fluid circulation grooves are opened on the bottom of the multi-porous culture plate body. Each group of culture fluid circulation grooves includes an input groove and a discharge groove. The channel sealing plate seals the bottom of the multi-porous culture plate body. The space formed by the input groove of each group of culture fluid circulation grooves and the channel sealing plate constitutes the input channel of the corresponding culture fluid circulation path. The space formed by the discharge groove of each group of culture fluid circulation grooves and the channel sealing plate constitutes the discharge channel of the corresponding culture fluid circulation path.

6. The high-throughput organoid culture liquid core device according to claim 3, characterized in that: The multi-well culture plate body is provided with coding marks distributed in a matrix, and each coding mark is located outside the bottom of the corresponding culture well.

7. The high-throughput organoid culture liquid core device according to claim 3, characterized in that: The multi-porous culture plate is square or round. If the multi-porous culture plate is square, the injection port and the discharge port of each culture fluid flow path are respectively arranged on the two side walls in the width direction of the multi-porous culture plate, and the straight line of each row of culture holes is parallel to the length direction of the multi-porous culture plate.

8. The high-throughput organoid culture liquid core device according to claim 1, characterized in that: The bottom of the transwell chamber is sealed, and a plurality of semipermeable membrane windows uniformly distributed along the circumferential direction are provided on the side wall of the transwell chamber.

9. The high-throughput organoid culture liquid core device according to claim 8, characterized in that: The transwell chamber comprises a transwell chamber body, a semipermeable membrane and a fixed sleeve. The bottom of the transwell chamber body is sealed, the bottom of the transwell chamber body is disc-shaped, the side wall of the transwell chamber body is provided with a plurality of first through grooves evenly distributed along the circumferential direction, the lower part of the side wall of the transwell chamber body is annular, the first through grooves evenly divide the lower part of the side wall of the transwell chamber body into mutually isolated inner pressure plates, the outer wall of the bottom of the transwell chamber body is connected to the inner wall of each inner pressure plate, the semipermeable membrane is annular, the fixed sleeve is annular, and the fixed sleeve is provided with a plurality of second through grooves evenly distributed along the circumferential direction. The two through grooves cooperate with the corresponding first through grooves, a snap ring is provided at the bottom of the inner wall of the fixing sleeve, the outer wall of the snap ring is fitted with the bottom of the inner wall of the fixing sleeve, a plurality of protrusions evenly distributed along the circumferential direction are provided on the top of the snap ring, and a snap groove for fixing the semipermeable membrane is provided between the two ends of each protrusion and the inner wall of the fixing sleeve, the semipermeable membrane is fitted to the inner wall of the fixing sleeve, and the lower end of the semipermeable membrane is inserted into each snap groove, the lower part of the side wall of the transwell chamber body is embedded in the semipermeable membrane, the bottom of each inner pressure plate is located on the top of the snap ring, the bottom of the transwell chamber body is embedded in the snap ring, the bottom of the transwell chamber body is interference fit with the snap ring, and each first through groove, the corresponding second through groove and the semipermeable membrane form a semipermeable membrane window.

10. The high-throughput organoid culture liquid core device according to claim 9, characterized in that: The first through groove extends to the top of the side wall of the transwell chamber body. The side wall of the transwell chamber body is composed of multiple internal pressure plates. Arc-shaped connecting strips are provided between adjacent internal pressure plates. Each connecting strip is connected to the middle part of two adjacent internal pressure plates. The top of the fixed sleeve is connected to each connecting strip by a snap buckle.

11. The high-throughput organoid culture liquid core device according to claim 1, characterized in that: The upper cover module includes an upper cover body and a sealing ring. The upper cover body covers the top of the multi-porous culture plate. The bottom of the outer wall of the upper cover body covering the sealing part is provided with a sealing groove that cooperates with the sealing ring, and the sealing ring is sleeved in the sealing groove.

12. The high-throughput organoid culture liquid core device according to claim 1, characterized in that: The outer side wall of the upper cover body is provided with an anti-slip tooth structure, and the corresponding position of the outer side wall of the multi-porous culture plate is provided with an anti-slip tooth structure.

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