3D printing organoid culture scaffold and organoid culture method
By using 3D-printed organoid culture scaffolds with microfluidic and microporous structures, the problem of insufficient nutrient supply to the central cells of organoids has been solved, enabling long-term growth and functional simulation of organoids, which is suitable for medical and scientific research applications.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- OTRIXELL BIOTECHNOLOGY(SUZHOU) CO LTD
- Filing Date
- 2023-03-28
- Publication Date
- 2026-05-12
AI Technical Summary
In existing organoid culture technologies, the central and surface cells do not come into contact with enough culture medium during the differentiation process, causing the organoids to stop growing after differentiating to a certain extent and thus failing to effectively mimic the function of human organs.
The organoid culture scaffold, manufactured using 3D printing technology, consists of multiple branches with interconnected microchannels and micropores inside. Culture medium is delivered through inlets and outlets to ensure full contact between cells and the culture medium, providing nutrient supply to the organoid center.
This improves the culture efficiency of organoids, allowing them to grow for longer periods and enhancing their ability to mimic human organ functions, making them suitable for medical research and scientific studies.
Smart Images
Figure CN116286357B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of organoid culture, and more particularly to a 3D-printed organoid culture scaffold and an organoid culture method. Background Technology
[0002] Over the past decade, the field of stem cell research has made tremendous progress, with a significant breakthrough being the rapid development of organoid systems from scratch. Organoid culture represents an advanced stage of 3D cell culture, characterized by the differentiation of cells into cell populations with different functions, capable of mimicking the functions of human organs to a certain extent. It has already begun to show promise in the medical field, such as in tumor drug sensitivity testing and high-throughput drug screening, demonstrating broad research and application potential. The ultimate goal of organoid development is to culture human organs in vitro, thereby replacing allogeneic transplantation in the market, possessing immense medical value and market prospects.
[0003] However, to date, organoids will "die" after being cultured to a certain extent, and current scientific progress is far from achieving the goal of culturing organs in vitro. Simply put, during the growth process, cells differentiate and proliferate, leading to the formation of necrotic cores. This is because growth-promoting substances such as culture media cannot penetrate to the center of the organoid for material exchange. In contrast, the human body has a vascular network that covers the entire body, providing nutrients and removing waste from various tissues and internal organs, thus maintaining human physiological functions.
[0004] Currently, the mainstream organoid culture methods internationally are mainly divided into two types: conventional culture plate culture and high-throughput microfluidic (i.e., organ-chip) culture. Conventional culture plate culture generally uses Matrigel matrix gel and special culture medium loaded with growth factors to culture cells. This method requires manual periodic replacement of the culture medium. High-throughput microfluidic culture generally places cells in chambers in the middle of the microfluidic channels. Culture medium containing substances required for organoid growth and differentiation is pumped into the cell growth chamber within the microfluidic chip through one or more inlets and outlets using a peristaltic pump, achieving automatic culture medium replacement. In addition to Matrigel matrix gel, microspheres and other hydrogel materials can also be used in the chambers to create a 3D microenvironment to support cell growth.
[0005] All of the methods mentioned above present a significant challenge: during the differentiation and growth of cells into organoids, the central cells receive less culture medium (i.e., material exchange) compared to the surface cells. This leads to cell death starting from the center after a certain stage of growth, ultimately making it difficult for organoids to continue growing after differentiation and thus preventing them from further mimicking the functions of human organs. Summary of the Invention
[0006] The purpose of this invention is to provide a 3D-printed organoid culture scaffold and an organoid culture method. By using this structure and method, the contact between the central cells and the culture medium can be increased during cell culture, thereby improving the culture effect.
[0007] To achieve the above objectives, the technical solution adopted by the present invention is: a 3D printed organoid culture scaffold, comprising a main body, wherein the main body has microchannels, one end of the main body has an inlet and the other end has an outlet, and the inlet and outlet are respectively connected to the microchannels;
[0008] The main body includes a main branch and a support. The main branch is in two sets. The microchannel is disposed in the support. The two ends of the support are connected to the inlet and outlet through the two sets of main branches respectively. The microchannel is connected to the inlet and outlet through the main branch.
[0009] The outer wall of the support is provided with multiple sets of micropores at intervals, and the micropores are connected to the microchannels.
[0010] The support is composed of multiple sets of branches connected together, and multiple sets of interconnected culture holes are formed between adjacent branches;
[0011] Each group of branches has microchannels extending along the branches inside, and the microchannels in multiple groups of branches are interconnected.
[0012] The micropores are spaced apart on the outer wall of the branch, and the micropores connect the microchannels to the culture wells.
[0013] In the above technical solution, the main branch is provided with a main channel, and the microchannel is connected to the inlet and outlet respectively through the main channel.
[0014] In the above technical solution, the support is further provided with an outer sleeve, the inner part of the outer sleeve is provided with a cavity, the outer end of the support is connected to the inner wall of the cavity, and the main branches are arranged at both ends of the outer sleeve.
[0015] The outer wall of the jacket is provided with multiple sets of through holes that communicate with the cavity, and the diameter of the through holes is larger than the outer diameter of the branch.
[0016] In the above technical solution, the branches at the connection between the support and the outer jacket are provided with end micro-holes, and the end micro-holes are connected to the outer surface of the outer jacket.
[0017] In the above technical solution, the support includes multiple sets of planar frames and multiple sets of upright frames, with the multiple sets of planar frames arranged at intervals, and each set of upright frames connecting adjacent planar frames;
[0018] The culture well is formed between adjacent planar frames and adjacent upright frames;
[0019] The branches include a first branch, a second branch, and a third branch;
[0020] The planar frame includes multiple sets of first branches and second branches that are interconnected with each other. Each set of first branches and second branches is provided with microchannels, and the microchannels in the first branch are connected to the microchannels in the second branch.
[0021] The support frame includes multiple sets of third branches, the ends of which are connected to the planar frame, and the microchannels in the third branches are connected to the microchannels in the first and / or second branches.
[0022] In the above technical solution, the pore size of the culture well is 100 micrometers to 1000 micrometers, the diameter of the microchannel is 2 micrometers to 500 micrometers, and the diameter of the micropore is less than or equal to the diameter of the microchannel.
[0023] In the above technical solution, the outer shell is a spherical structure or a near-spherical structure.
[0024] In the above technical solution, the support is a planar frame, which includes multiple sets of branches spaced apart from top to bottom. The two ends of each set of branches are connected to the main branches on both sides, and the microchannels in the branches are connected to the inlet and outlet through the main branches on both sides.
[0025] This invention also provides a method for organoid culture, comprising the following steps:
[0026] ① Use a 3D printer to print the above-mentioned 3D printed organoid culture scaffold;
[0027] ② After the culture scaffold prepared in step ① is fully swollen in the buffer solution, it is placed into the chamber of the microfluidic chip;
[0028] ③ Use a dropper to drip the cells and culture medium to be cultured onto the culture scaffold through the feed port of the microfluidic chip, so that the culture medium fully immerses the culture scaffold;
[0029] ④ Place the microfluidic chip containing the culture scaffold and cells into a carbon dioxide incubator for culture, and use a peristaltic pump to deliver and replace the culture medium and growth promoters in the microfluidic chip;
[0030] In step ④, the inlet of the culture scaffold is connected to the inlet of the microfluidic chip, and the outlet of the culture scaffold is connected to the outlet of the microfluidic chip. The microfluidic chip fixes the culture scaffold in the chamber.
[0031] The culture medium flows into the inlet of the culture scaffold through the inlet of the microfluidic chip, and is further delivered into the culture wells through microchannels and micropores, and then to any position on the culture scaffold for cell absorption and culture. The culture medium then flows out of the microfluidic chip through the outlet of the culture scaffold, realizing the exchange of substances on the culture scaffold.
[0032] In the above technical solution, the manufacturing process of the culture scaffold in step ① is as follows:
[0033] a. Use a 3D printer to print the culture scaffold in one piece to obtain a pre-formed culture scaffold;
[0034] b. The pre-formed culture scaffold is cleaned, dried, and sterilized in sequence to obtain the formed culture scaffold.
[0035] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art:
[0036] 1. In this invention, the scaffold is composed of multiple interconnected branches, and the branches are provided with interconnected microchannels. The outer wall of the branches is provided with micropores that communicate with the microchannels. The culture medium is transported and exchanged through the inlet and outlet. By using the microchannels and micropores, the cells in the center of the organoid can be supplied with culture medium and other growth-promoting substances, so as to culture the organoid for a longer period of time and make the culture more durable, thereby improving the culture effect.
[0037] 2. In this invention, multiple interconnected culture wells are formed between adjacent branches, which provides an ideal 3D growth microenvironment for the cultured cells. Furthermore, the culture wells can be interconnected, and the microchannels and micropores act as capillaries to provide culture medium, so that all parts of the organoid can be covered by the culture medium, effectively improving the culture effect. Attached Figure Description
[0038] Figure 1 This is a schematic diagram of the structure in Embodiment 1 of the present invention;
[0039] Figure 2 yes Figure 1 A schematic diagram of the cross-sectional structure;
[0040] Figure 3 This is a partial structural diagram of the culture scaffold after it has been cut open in Embodiment 1 of the present invention;
[0041] Figure 4 This is a partial structural schematic diagram of the bracket in Embodiment 1 of the present invention;
[0042] Figure 5 This is a schematic diagram of the structure in Embodiment 2 of the present invention.
[0043] Among them: 1. Microchannel; 2. Inlet; 3. Outlet; 4. Main branch; 5. Support; 6. Micropore; 7. Branch; 8. Culture hole; 9. Main channel; 10. Outer shell; 11. Through hole; 12. End micropore; 13. Planar frame; 14. Stand; 15. First branch; 16. Second branch; 17. Third branch. Detailed Implementation
[0044] The present invention will be further described below with reference to the accompanying drawings and embodiments:
[0045] Example 1: See Figures 1-4 As shown, a 3D-printed organoid culture scaffold includes a main body with a microfluidic channel 1 inside. One end of the main body has an inlet 2 and the other end has an outlet 3, which are respectively connected to the microfluidic channel. The inlet and outlet are also connected to the microfluidic chip channel, so that the microfluidic chip can provide the main body with culture medium and other growth-promoting substances.
[0046] The main body includes a main branch 4 and a support 5. The main branches are in two sets. The microchannel is disposed in the support. The two ends of the support are connected to the inlet and outlet through the two sets of main branches respectively. The microchannel is connected to the inlet and outlet through the main branches.
[0047] The outer wall of the support is provided with multiple sets of micropores 6 at intervals, and the micropores are connected to the microchannels.
[0048] The support is composed of multiple sets of branches 7 connected to each other, and multiple sets of interconnected culture holes 8 are formed between adjacent branches;
[0049] Each group of branches has microchannels extending along the branches inside, and the microchannels in multiple groups of branches are interconnected.
[0050] The micropores are spaced apart on the outer wall of the branch, and the micropores connect the microchannels to the culture wells.
[0051] This invention also provides a method for organoid culture, comprising the following steps:
[0052] ① Use a 3D printer to print the above-mentioned 3D printed organoid culture scaffold;
[0053] ② After fully swelling the culture scaffold prepared in step ① in a buffer solution, place it into the chamber of the microfluidic chip. The buffer solution is a solution that can resist pH changes when small amounts of acid, alkali, or water are added. pH buffering systems play a crucial role in maintaining the normal pH value and physiological environment of organisms. Most cells can only function within a very narrow pH range and require a buffer system to resist pH changes during metabolism. Swelling occurs because the material expands after being soaked in water, and after reaching a certain point, it stops expanding, allowing it to stabilize before proceeding with subsequent steps.
[0054] ③ Use a dropper to drip the cells and culture medium to be cultured onto the culture scaffold through the feed port of the microfluidic chip, so that the culture medium fully immerses the culture scaffold;
[0055] ④ Place the microfluidic chip containing the culture scaffold and cells into a carbon dioxide incubator for culture, and use a peristaltic pump to deliver and replace the culture medium and growth promoters in the microfluidic chip;
[0056] In step ④, the inlet of the culture scaffold is connected to the inlet of the microfluidic chip, and the outlet of the culture scaffold is connected to the outlet of the microfluidic chip. The microfluidic chip fixes the culture scaffold in the chamber.
[0057] The culture medium flows into the inlet of the culture scaffold through the inlet of the microfluidic chip, and is further delivered to any position on the culture scaffold through microchannels and micropores for cell absorption and culture. It then flows out of the microfluidic chip through the outlet of the culture scaffold, realizing the exchange of substances on the culture scaffold.
[0058] In this invention, during organoid culture, the microfluidic chip replaces or exchanges the culture medium in the chamber through inlets and outlets, eliminating the need for manual medium replacement. The cells to be cultured are enclosed in a three-dimensional microenvironment constructed by a 3D-printed scaffold, where they amplify and differentiate by absorbing growth-promoting substances from the culture medium. Under normal conditions, the chamber of the microfluidic chip is filled with culture medium. In the initial cell amplification phase, due to the small number of cells, almost all cells can access the culture medium, allowing for normal amplification and differentiation, thus facilitating organoid culture. During cell amplification, outer cells envelop the inner cells, differentiating layer by layer outwards. The outer cells can easily exchange substances with the culture medium within the microfluidic chip's chamber. In conventional structures, cells near the center are enclosed by outer cells, making it difficult for external culture medium to penetrate the inner layer. This leads to apoptosis of inner cells after a certain growth stage, preventing further organoid growth. In this invention, the main body is composed of multiple interconnected branches, each branch containing microchannels that communicate with other branches. Each branch also has multiple sets of micropores connected to these microchannels. This allows for nutrient exchange between the inner cells and the culture medium through these micropores. Simultaneously, cells near the micropores can also contact the culture medium, providing nutrients to these cells. Furthermore, during cell proliferation and differentiation, cells enter the micropores and microchannels, partially blocking some of the micropores and sections of the microchannels. Because of the large number of micropores and the interconnectedness of the microchannels, the unblocked microchannels and micropores can also provide culture medium to adjacent cells, maximizing organoid growth and facilitating subsequent scientific or medical research, thus achieving better research and medical outcomes.
[0059] See Figure 2 As shown, the main branch has a main channel 9, and the microchannels are connected to the inlet and outlet via the main channel. The main channel is used to facilitate the exchange of microfluidic culture medium. The outer wall of the main branch has microchannels that communicate with the main channel, so that the culture medium transported in the main channel can also be delivered to the outside through the microchannels on the main branch.
[0060] See Figures 1-3 As shown, the support is further provided with an outer sleeve 10, the inner part of the outer sleeve is provided with a cavity, the outer end of the support is connected to the inner wall of the cavity, and the main branches are located at both ends of the outer sleeve.
[0061] The outer wall of the jacket is provided with multiple sets of through holes 11 that communicate with the cavity, and the diameter of the through holes is larger than the outer diameter of the branch.
[0062] In this embodiment, the cells to be cultured are dripped into the scaffold inside the outer sheath through the perforations. This allows the outer sheath to restrict cell culture, ensuring that the cells adhere as closely as possible to the culture wells within the scaffold and remain within the outer sheath, thus limiting the developmental space of the organoids. Simultaneously, the perforations allow external culture medium to enter the outer sheath and contact the cells, providing nutrients and facilitating nutrient exchange. Furthermore, the use of 3D printing technology allows for convenient printing of the 3D-printed scaffold size based on different organoid types and required dimensions, enabling personalized fabrication for different organoids.
[0063] The outer casing is spherical or near-spherical, allowing for maximum contact with the culture medium at various points during organoid growth, thus increasing the contact area and improving culture efficiency. In this invention, the microfluidic chip chamber housing the culture scaffold has inlet / outlet structures at both ends to secure it within the chamber and connect it to the inlet / outlet on the microfluidic chip. The microfluidic inlet / outlet is connected to the peristaltic pump's piping. The upper end of the microfluidic chip has an inlet connected to the chamber, facilitating the dripping of cells and culture medium into the chamber.
[0064] See Figure 1 , 3 As shown, each branch at the connection between the support and the outer cover is provided with an end microhole 12, and the end microhole is connected to the outer surface of the outer cover.
[0065] In this embodiment, the end micropores are connected to the outer surface of the outer sheath. This allows the culture medium to still enter the microchannels through the end micropores during cell proliferation and differentiation, even if some microchannels are blocked. This ensures that the micropores provide the inner cells with the culture medium for nutrient replacement.
[0066] See Figures 2-4 As shown, the support includes multiple sets of planar frames 13 and multiple sets of upright frames 14. The multiple sets of planar frames are arranged at intervals, and each set of upright frames connects adjacent planar frames.
[0067] The culture well is formed between adjacent planar frames and adjacent upright frames;
[0068] The branches include a first branch 15, a second branch 16, and a third branch 17;
[0069] The planar frame includes multiple sets of intersecting first branches and second branches. Each set of first branches and second branches is provided with microchannels, and the microchannels in the first branch are connected to the microchannels in the second branch. The microchannels in the first branch and the microchannels in the second branch are connected at their intersection.
[0070] The support frame includes multiple sets of third branches. The ends of the third branches are connected to the planar frame, and the microchannels within the third branches are connected to the microchannels within the first and / or second branches. In this embodiment, the ends of the third branches are connected to the junctions of the first and second branches, allowing the microchannels within the first, second, and third branches to communicate at these junctions. Alternatively, the third branches may only connect to the first or second branches, thereby enabling communication between the microchannels of the third branch and the microchannels of the first or second branch.
[0071] In this embodiment, multiple planar frames are connected using uprights, thus forming a three-dimensional support. The first, second, and third branches are interconnected, and the internal microchannels are interconnected. This way, even if the micropores and microchannels in some areas are blocked, there will still be many unblocked pathways. These unblocked pathways are used to provide culture medium to the corresponding unblocked micropores, providing channels for nutrient exchange, thereby promoting better development of organoids.
[0072] Simultaneously, the adjacent planar supports and adjacent upright supports form the culture wells. This allows cells within the culture wells to receive culture medium from the micropores on the adjacent planar and upright supports. In other words, the organoid tissue within the culture wells has culture medium delivery channels around its perimeter, functioning like capillaries to supply nutrients and facilitate substance exchange to the inner cells, ensuring better growth and development of the organoid. Furthermore, the scaffold also provides cells with a supportive 3D growth microenvironment.
[0073] The culture well has a pore size of 100 micrometers to 1000 micrometers, the microchannel has a diameter of 2 micrometers to 500 micrometers, and the diameter of the micropore is less than or equal to the diameter of the microchannel.
[0074] The culture wells have a moderate pore size, providing a 3D culture microenvironment with supporting structures for cells. Micropores around the wells provide culture medium, ensuring that cells within each well can contact the medium for nutrient exchange. The stems employ a hollow tetrahedral structure, with micropores located on at least the opposite sidewalls of the main stem, preferably on all four sides. This allows for the supply of culture medium to the corresponding wells through these micropores, ensuring uniform nutrient supply and exchange for cells within each well, thus promoting better organoid growth.
[0075] In step ①, the manufacturing process of the culture scaffold is as follows:
[0076] a. Use a 3D printer to print the culture scaffold in one piece to obtain a pre-formed culture scaffold;
[0077] b. The pre-formed culture scaffold is cleaned, dried, and sterilized in sequence to obtain the formed culture scaffold.
[0078] In this invention, the main body and the outer shell are 3D printed as a single piece, which can be made of biodegradable materials or non-biodegradable materials.
[0079] Among them, the non-degradable materials are preferably one or more of the following: titanium, titanium alloy, tantalum metal, stainless steel, and cobalt-based alloy.
[0080] The preferred biodegradable materials are: polyethylene glycol and its polymers, polyethylene glycol diacrylate (PEGDA), collagen, methacrylated collagen, methacrylated gelatin (GELMA), polyvinyl alcohol, methacrylated polyvinyl alcohol (PVAMA), polycaprolactone, methacrylated polycaprolactone (PCLMA), methacrylated chitosan (CSMA), polylactic acid, hydroxyapatite, tricalcium phosphate, and nanoclay.
[0081] Example 2: See Figure 5 As shown, a 3D-printed organoid culture scaffold has a structure that is basically similar to that of Embodiment 1, except that the scaffold is a planar frame, which includes three sets of branches spaced apart from top to bottom. The two ends of each set of branches are connected to the main branches on both sides, and the microchannels in the branches are connected to the inlet and outlet through the main branches on both sides.
[0082] In this embodiment, the microchannels of the three sets of branches form a three-channel structure, a quasi-three-layer structure, which spatially constitutes a porous network. This allows the culture medium to flow out through the micropores on the outer surface of the branches, providing nutrients for cell absorption. Compared to the structure in Embodiment 1, this structure is more conducive to constructing vascularized organoid structures and is better for suspension cells that do not require culture wells, as suspension cells do not need a 3D microenvironment for support structures and are suitable for planar frames.
[0083] In the description, it should be understood that the terms "upper," "lower," "top," "bottom," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. In the description of the invention, "a plurality of" means two or more, unless otherwise explicitly specified.
[0084] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium. For instance, the two components can be mechanically connected by contact or abutting; they can also be directly hooked or connected by an intermediate medium; or they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.
Claims
1. A 3D-printed organoid culture scaffold, characterized in that: It includes a main body, which has microchannels, with an inlet at one end and an outlet at the other end, and the inlet and outlet are respectively connected to the microchannels; The main body includes a main branch and a support. The main branch is in two sets. The microchannel is disposed in the support. The two ends of the support are connected to the inlet and outlet through the two sets of main branches respectively. The microchannel is connected to the inlet and outlet through the main branch. The outer wall of the support is provided with multiple sets of micropores at intervals, and the micropores are connected to the microchannels. The scaffold is composed of multiple sets of branches connected together, and multiple sets of interconnected culture wells are formed between adjacent branches. The culture wells and the scaffold provide a 3D growth microenvironment with a supporting structure for the cells. The support includes multiple sets of planar frames and multiple sets of upright frames. The multiple sets of planar frames are arranged at intervals, and each set of upright frames connects adjacent planar frames. The culture well is formed between adjacent planar frames and adjacent upright frames; Each group of branches has microchannels extending along the branches inside, and the microchannels in multiple groups of branches are interconnected. The micropores are spaced apart on the outer wall of the branch, and the micropores connect the microchannels to the culture wells. The micropores provide culture medium to the cells in the culture wells, and the cells are given nutrients and exchange substances through the culture medium.
2. The 3D-printed organoid culture scaffold according to claim 1, characterized in that: The main branch is provided with a main channel, and the microchannels are connected to the inlet and outlet respectively through the main channel.
3. The 3D-printed organoid culture scaffold according to claim 1, characterized in that: The support is also provided with an outer sleeve, the inner part of which has a cavity. The outer end of the support is connected to the inner wall of the cavity, and the main branches are located at both ends of the outer sleeve. The outer wall of the jacket is provided with multiple sets of through holes that communicate with the cavity, and the diameter of the through holes is larger than the outer diameter of the branch.
4. The 3D-printed organoid culture scaffold according to claim 3, characterized in that: Each branch at the connection point between the support and the outer cover is provided with an end micro-hole, which is connected to the outer surface of the outer cover.
5. The 3D-printed organoid culture scaffold according to claim 1, characterized in that: The branches include a first branch, a second branch, and a third branch; The planar frame includes multiple sets of first branches and second branches that are interconnected with each other. Each set of first branches and second branches is provided with microchannels, and the microchannels in the first branch are connected to the microchannels in the second branch. The support frame includes multiple sets of third branches, the ends of which are connected to the planar frame, and the microchannels in the third branches are connected to the microchannels in the first and / or second branches.
6. The 3D-printed organoid culture scaffold according to claim 1, characterized in that: The pore size of the culture well is 100 micrometers to 1000 micrometers, the diameter of the microchannel is 2 micrometers to 500 micrometers, and the diameter of the micropore is less than or equal to the diameter of the microchannel.
7. The 3D-printed organoid culture scaffold according to claim 3, characterized in that: The outer casing has a spherical or near-spherical structure.
8. The 3D-printed organoid culture scaffold according to claim 1, characterized in that: The support is a planar frame, which includes multiple sets of branches spaced apart from top to bottom. The two ends of each set of branches are connected to the main branches on both sides, and the microchannels in the branches are connected to the inlet and outlet through the main branches on both sides.
9. A method for culturing organoids, characterized in that: Includes the following steps: ① Print the 3D-printed organoid culture scaffold as described in any one of claims 1-8 using a 3D printer; ② After the culture scaffold prepared in step ① is fully swollen in the buffer solution, it is placed into the chamber of the microfluidic chip; ③ Use a dropper to drip the cells and culture medium to be cultured onto the culture scaffold through the feed port of the microfluidic chip, so that the culture medium fully immerses the culture scaffold; ④ Place the microfluidic chip containing the culture scaffold and cells into a carbon dioxide incubator for culture, and use a peristaltic pump to deliver and replace the culture medium and growth promoters in the microfluidic chip; In step ④, the inlet of the culture scaffold is connected to the inlet of the microfluidic chip, and the outlet of the culture scaffold is connected to the outlet of the microfluidic chip. The microfluidic chip fixes the culture scaffold in the chamber. The culture medium flows into the inlet of the culture scaffold through the inlet of the microfluidic chip, and is further delivered to any position on the culture scaffold through microchannels and micropores for cell absorption and culture. It then flows out of the microfluidic chip through the outlet of the culture scaffold, realizing the exchange of substances on the culture scaffold.
10. The organoid culture method according to claim 9, characterized in that: In step ①, the manufacturing process of the culture scaffold is as follows: a. Use a 3D printer to print the culture scaffold in one piece to obtain a pre-formed culture scaffold; b. The pre-formed culture scaffold is cleaned, dried, and sterilized in sequence to obtain the formed culture scaffold.