A high-throughput culture method for organoids
Through the combination of microfluidic control system and 3D printing system, high-throughput culture of tumor organoids is achieved, solving the problem of time-consuming, labor-intensive, uncontrollable and large individual differences in the organoid culture process in the existing technology, and realizing the organoids Fast, control and simulate the microenvironment of the human body, improving the efficiency of screening.
Patent Information
- Application Number
- CN202210869517.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-07-21
- Publication Date
- 2024-12-20
- Estimated Expiration
- 2042-07-21
AI Technical Summary
The culture process of existing tumor organoids is time-consuming and labor-intensive, and the operation is uncontrollable, resulting in uneven organoid sizes, inability to ensure consistency, large differences between individuals, and inability to simulate the microenvironment in the human body, and the growth and development time are long.
Using a high-throughput culture method combined with a microfluidic control system and a 3D printing system, cells are wrapped in dispersed phases, and uniformly sized organoid precursor droplets are prepared by using a microfluidic control system, and printed to the vascular bed microfluid through a 3D printing system. The chip-controlled culture is carried out in the culture room to achieve controllable and rapid organoid preparation and culture.
It achieves controllability and consistency of organoids, shortens growth and development time, simulates the microenvironment in the human body, and improves the efficiency and accuracy of screening drugs.
Smart Images
Figure CN115354030B_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of medical devices, and in particular to a high-throughput culture method for organoids. Background Art
[0002] The cultivation of primary organoids helps in the development of anti-cancer drugs and the precise drug screening for cancer patients.
[0003] Currently, the culture method of primary organoids such as tumor organoids is mainly divided into two steps: first, the preparation of tumor organoids, which usually depends on the technology of the experimenter; second, the experimenter manually transfers the prepared tumor organoids to the wells of the well plate for culture. This process is time-consuming and labor-intensive, the operation process is uncontrollable, the prepared organoids are uneven in size, the consistency of the organoids cannot be guaranteed, the differences between individual organoids are large, and the culture in the wells of the well plate cannot simulate the microenvironment in the human body, and the growth and development time of the organoids is relatively long. Summary of the invention
[0004] In order to overcome the shortcomings of the prior art, the present invention provides a high-throughput organoid culture method with good controllability, saving time and labor, ensuring the consistency of organoids, reducing the differences between individual organoids, simulating the microenvironment in the human body, and shortening the growth and development time of organoids.
[0005] The technical solution adopted by the present invention to solve its technical problem is:
[0006] A high-throughput organoid culture method comprises the following steps: S2, encapsulating cells in a dispersed phase 1; S4, respectively injecting the dispersed phase 1 and the dispersed phase 2 encapsulating the cells into the organoid precursor preparation area of the microfluidic system, wherein the dispersed phase 1 is sheared into organoid precursor droplets of uniform size by the dispersed phase 2, and the sheared organoid precursor droplets can sequentially enter the organoid droplet maturation area of the microfluidic system and can be matured into organoid spheres in the organoid droplet maturation area; S6, transporting the first organoid sphere in the organoid droplet maturation area to the print nozzle of the 3D printing system through the transport pipeline of the 3D printing system by the transport pump of the microfluidic system; S8, detecting whether there is an organoid sphere at the print nozzle by the organoid recognition detection unit of the 3D printing system, and when the organoid recognition detection unit detects that there is an organoid sphere at the print nozzle, the organoid recognition detection unit The measuring unit outputs the organoid signal to the control unit of the 3D printing system; S10, controlling the action of the driving unit of the 3D printing system by the control unit, and driving the printing nozzle to move downward to the bottom of the vascular bed culture chamber of the first vascular bed microfluidic chip by the driving unit. After staying for 1-2 seconds, the first organoid sphere will be adsorbed on the bottom of the vascular bed culture chamber of the first vascular bed microfluidic chip, so that the first organoid sphere is printed into the vascular bed culture chamber of the first vascular bed microfluidic chip. After completion, the printing nozzle is driven back to the initial position by the driving unit; S12, according to the method of step S6 to step S10, the remaining organoid spheres in the organoid droplet maturation area are printed into the vascular bed culture chambers of the remaining vascular bed microfluidic chips in sequence; S14, the organoid spheres in the vascular bed culture chambers of all vascular bed microfluidic chips are cultured.
[0007] As a preferred technical solution, the organoid precursor preparation area is a Y-shaped structure, which includes a first inlet pipe, a second inlet pipe and a connecting pipe, the first end of the first inlet pipe and the first end of the second inlet pipe are respectively connected to the first syringe and the second syringe, the first syringe is connected to a first injection pump, the second syringe is connected to a second injection pump, the second end of the first inlet pipe and the second end of the second inlet pipe are both connected to the first end of the connecting pipe, and the organoid droplet maturation area is an outlet pipe, and the second end of the connecting pipe is connected to the outlet pipe; the step S4 includes the following steps: S42, placing dispersed phase 1 and dispersed phase 2 in the first syringe and the second syringe respectively; S44, injecting dispersed phase 1 in the first syringe into the first inlet pipe through the first injection pump, and injecting dispersed phase 2 in the second syringe into the second inlet pipe through the second injection pump, in the connecting pipe, dispersed phase 1 is sheared into organoid precursor droplets of uniform size by dispersed phase 2, and the sheared organoid precursor droplets can enter the outlet pipe in sequence and can be matured into organoid spheres in the outlet pipe.
[0008] As a preferred technical solution, the organoid precursor preparation area is a T-shaped structure, which includes a first inlet pipe, a second inlet pipe and a connecting pipe, the first end of the first inlet pipe and the first end of the second inlet pipe are respectively connected to the first syringe and the second syringe, the first syringe is connected to a first injection pump, the second syringe is connected to a second injection pump, the second end of the first inlet pipe and the second end of the second inlet pipe are respectively connected to the first end of the connecting pipe, and the organoid droplet maturation area is an outlet pipe, and the second end of the connecting pipe is connected to the outlet pipe; the step S4 includes the following steps: S42, placing dispersed phase 1 and dispersed phase 2 in the first syringe and the second syringe, respectively; S44, injecting dispersed phase 1 in the first syringe into the first inlet pipe through the first injection pump, and injecting dispersed phase 2 in the second syringe into the second inlet pipe through the second injection pump, in the connecting pipe, dispersed phase 1 is sheared into organoid precursor droplets of uniform size by dispersed phase 2, and the sheared organoid precursor droplets can enter the outlet pipe in sequence and can be matured into organoid spheres in the outlet pipe.
[0009] As a preferred technical solution, the organoid precursor preparation area is a cross-shaped structure, which includes a first inlet pipe, two second inlet pipes arranged opposite to each other and a connecting pipe, the first inlet pipe and the connecting pipe are arranged opposite to each other, the first end of the first inlet pipe is connected to the first syringe, the second end of the first inlet pipe is connected to the first end of the connecting pipe, the first syringe is connected to a first injection pump, the first ends of the two second inlet pipes are respectively connected to two second syringes, the second ends of the two second inlet pipes are respectively connected to the first end of the connecting pipe, the two second syringes are respectively connected to two second injection pumps, and the organoid droplet maturation area is a The outlet pipe is connected to the second end of the connecting pipe; the step S4 comprises the following steps: S42, placing the dispersed phase 1 in the first syringe, and placing the dispersed phase 2 in two second syringes respectively; S44, injecting the dispersed phase 1 in the first syringe into the first inlet pipe through the first injection pump, and injecting the dispersed phase 2 in the two second syringes into the two second inlet pipes respectively through the two second injection pumps, in the connecting pipe, the dispersed phase 1 is injected into the dispersed phase 2 in the two second inlet pipes and sheared into organoid precursor droplets of uniform size, and the sheared organoid precursor droplets can enter the outlet pipe in sequence and can be matured into organoid spheres in the outlet pipe.
[0010] As a preferred technical solution, in step S2, the cells are primary tumor tissue cells, stem cells or cell lines.
[0011] As a preferred technical solution, the material of the dispersed phase 1 includes matrix gel, collagen or gelatin.
[0012] As a preferred technical solution, the material of the dispersed phase 2 includes fluorine oil or vegetable oil.
[0013] As a preferred technical solution, in step S4, the method of maturing the organoid precursor droplets into organoid spheres includes temperature crosslinking, enzymatic crosslinking, ionic crosslinking or photocrosslinking.
[0014] The beneficial effects of the present invention are as follows: the high-throughput organoid culture method provided by the present invention completes the preparation of organoids through a microfluidic system, and through the combination of the microfluidic system and the 3D printing system, the organoids can be simply, quickly and accurately transferred to the bottom of the vascular bed culture chamber of the vascular bed microfluidic chip for culture by printing. The process has good controllability, saves time and effort, has good stability, and the organoids are uniform in size, which ensures the consistency of the organoids and reduces the differences between individual organoids. At the same time, printing the organoids into the vascular bed culture chamber of the vascular bed microfluidic chip for culture can simulate the microenvironment in the human body, and a complete vascular network can be constructed inside the organoid within a short time (such as 3-5 days), shortening the growth and development time of the organoids and improving the efficiency and accuracy of drug screening. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] The present invention is further described below in conjunction with the accompanying drawings and embodiments.
[0016] Figure 1 It is a flowchart of a high-throughput culture method of organoids provided by one embodiment of the present invention;
[0017] Figure 2 yes Figure 1 A schematic diagram of a process for high-throughput culture of organoids is shown;
[0018] Figure 3 It is a schematic diagram of the extraction of primary tumor tissue cells from primary tumor tissue;
[0019] Figure 4 is a schematic diagram of the structure of a first syringe and a second syringe;
[0020] Figure 5 This is a plan view of the organoid precursor preparation area and the organoid droplet maturation area of the first scheme of the microfluidic system;
[0021] Figure 6 It is a plan view of the organoid precursor preparation area and the organoid droplet maturation area of the second scheme of the microfluidic system;
[0022] Figure 7 This is a schematic plan view of the organoid precursor preparation area and the organoid droplet maturation area of the third scheme of the microfluidic system;
[0023] Figure 8 It is a plan view of the organoid precursor preparation area and the organoid droplet maturation area of the fourth scheme of the microfluidic system;
[0024] Fig. 9 This is a schematic diagram of the effect of culturing organoid spheroids in the vascular bed culture chamber of the vascular bed microfluidic chip. DETAILED DESCRIPTION
[0025] The following will clearly and completely describe the concept, specific structure and technical effects of the present invention in combination with the embodiments and drawings, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by technicians in this field without creative work are all within the scope of protection of the present invention. In addition, all the connection / connection relationships involved in the patent do not refer to the direct connection of components, but refer to the formation of a better connection structure by adding or reducing connection accessories according to the specific implementation situation. The various technical features in the invention can be combined interchangeably without conflicting with each other.
[0026] Please refer to Figure 1 and Figure 2 An embodiment of the present invention provides a high-throughput organoid culture method, comprising the following steps:
[0027] S2. Encapsulate the cells in dispersed phase 1.
[0028] In step S2, the cells are primary tumor tissue cells extracted from primary tumor tissue, such as Figure 3 It is understandable that the cells may also be, for example, stem cells, cell lines, etc., and the type and quantity of cells may be set according to actual conditions.
[0029] In this embodiment, the cell density encapsulated in the dispersed phase 1 is 1×10 7 Cells / ml. The cell density compatible with the present invention includes but is not limited to 1×10 5 / mL to 1×10 9 / ml, which can be flexibly adjusted according to culture requirements and cell activity.
[0030] The material of the dispersed phase 1 includes matrix gel. It can be understood that the material of the dispersed phase 1 can also include natural materials such as collagen, gelatin, etc. The material of the dispersed phase 1 can also include synthetic materials, which can be set according to actual conditions.
[0031] S4, the dispersed phase 1 and the dispersed phase 2 containing cells are respectively injected into the organoid precursor preparation area of the microfluidic system, and the dispersed phase 1 is sheared into organoid precursor droplets 100 of uniform size by the dispersed phase 2. The outside of the organoid precursor droplet 100 is partially wrapped with the dispersed phase 2. The sheared organoid precursor droplets 100 can sequentially enter the organoid droplet maturation area of the microfluidic system and can be matured into organoid spheres 102 in the organoid droplet maturation area, and the organoid is prepared.
[0032] In step S4, the material of the dispersed phase 2 includes fluorinated oil. Fluorinated oil can better produce droplet phenomenon and has better shear force, so that the dispersed phase 1 encapsulating cells forms organoid precursor droplets 100 of uniform size. It can be understood that the dispersed phase 2 can also include materials such as vegetable oil that have good cell compatibility and are incompatible with water, and can be set according to actual conditions.
[0033] Methods for maturing the organoid precursor droplets 100 into organoid spheres 102 include temperature crosslinking, enzymatic crosslinking, ion crosslinking, photocrosslinking, and the like.
[0034] In this embodiment, the organoid precursor preparation area is a T-shaped structure. Figure 5 The T-shaped structure includes a first inlet pipe 42, a second inlet pipe 43 and a connecting pipe 44. The first end of the first inlet pipe 42 and the first end of the second inlet pipe 43 are respectively connected to the first syringe 32 (see Figure 4 ), the second syringe 34 (see Figure 4 ), the first syringe 32 is connected to the first injection pump, the second syringe 34 is connected to the second injection pump, and the second end of the first inlet pipe 42 and the second end of the second inlet pipe 43 are respectively connected to the first end of the connecting pipe 44. The axis of the first inlet pipe 42 and the axis of the connecting pipe 44 are located on the same horizontal line, and the second inlet pipe 43 is vertically arranged with the first inlet pipe 42 and the connecting pipe 44. The second inlet pipe 43 is located above the first inlet pipe 42 and the connecting pipe 44. The organoid droplet maturation zone is an outlet pipe 50, and the second end of the connecting pipe 44 is connected to the outlet pipe 50. The connecting pipe 44 and the outlet pipe 50 are connected as a whole.
[0035] Step S4 specifically includes the following steps:
[0036] S42, placing dispersed phase 1 and dispersed phase 2 in the first syringe 32 and the second syringe 34 respectively.
[0037] S44, the dispersed phase 1 in the first syringe 32 is injected into the first inlet pipe 42 by the first syringe pump, and the dispersed phase 2 in the second syringe 34 is injected into the second inlet pipe 43 by the second syringe pump. In the connecting pipe 44, the dispersed phase 1 is sheared into organoid precursor droplets 100 of uniform size by the dispersed phase 2. The sheared organoid precursor droplets 100 can enter the outlet pipe 50 in sequence and can be matured into organoid spheres 102 in the outlet pipe 50. Figure 2 and Figure 5 shown.
[0038] In this embodiment, the flow rate of dispersed phase 1 is generally set to 20 μL / min, and the flow rate of dispersed phase 2 is generally set to 120 μL / min. The present invention can adjust the flow rates and flow rate ratio of the two dispersed phases according to needs, thereby adjusting the shape of the generated organoid precursor droplets (e.g., circular or elliptical with different lengths), the spacing of the organoid precursor droplets 100, and the overall speed of preparing the organoid precursor droplets 100.
[0039] In the first alternative, if Figure 6 As shown in Figure 2, the organoid precursor preparation area is also a T-shaped structure. Figure 5 The difference from the organoid precursor preparation area is that the axis of the first inlet pipe 42 and the axis of the second inlet pipe 43 are located on the same horizontal line, the connecting pipe 44 is vertically arranged to the first inlet pipe 42 and the second inlet pipe 43, and the connecting pipe 44 is located below the first inlet pipe 42 and the second inlet pipe 43.
[0040] In the second alternative, if Figure 7 As shown, the organoid precursor preparation area is a Y-shaped structure. The Y-shaped structure includes a first inlet pipe 42, a second inlet pipe 43 and a connecting pipe 44. The first end of the first inlet pipe 42 and the first end of the second inlet pipe 43 are respectively connected to the first syringe 32 and the second syringe 34, and the second end of the first inlet pipe 42 and the second end of the second inlet pipe 43 are both connected to the first end of the connecting pipe 44. The first inlet pipe 42 and the second inlet pipe 43 form a V-shaped structure, and the connecting pipe 44 is located below the first inlet pipe 42 and the second inlet pipe 43. The organoid droplet maturation area is an outlet pipe 50, and the second end of the connecting pipe 44 is connected to the outlet pipe 50. The connecting pipe 44 and the outlet pipe 50 are connected as a whole.
[0041] Step S4 includes the following steps:
[0042] S42, placing dispersed phase 1 and dispersed phase 2 in the first syringe 32 and the second syringe 34 respectively;
[0043] S44. The dispersed phase 1 in the first syringe 32 is injected into the first inlet pipe 42 through the first injection pump, and the dispersed phase 2 in the second syringe 34 is injected into the second inlet pipe 43 through the second injection pump. In the connecting pipe 44, the dispersed phase 1 is sheared into organoid precursor droplets 100 of uniform size by the dispersed phase 2. The sheared organoid precursor droplets 100 can enter the outlet pipe 50 in sequence and can be matured into organoid spheres 102 in the outlet pipe 50.
[0044] In the third alternative, if Figure 8 As shown, the organoid precursor preparation area is a cross-shaped structure. The cross-shaped structure includes a first inlet pipe 42, two second inlet pipes 43 arranged opposite to each other, and a connecting pipe 44. The first inlet pipe 42 and the connecting pipe 44 are arranged opposite to each other, and the first end of the first inlet pipe 42 is connected to the first syringe 32, and the second end of the first inlet pipe 42 is connected to the first end of the connecting pipe 44. The first syringe 32 is connected to a first injection pump. The first ends of the two second inlet pipes 43 are respectively connected to the two second syringes 34, and the second ends of the two second inlet pipes 43 are respectively connected to the first end of the connecting pipe 44. The two second syringes 34 are respectively connected to two second injection pumps. The organoid droplet maturation area is an outlet pipe 50, and the second end of the connecting pipe 44 is connected to the outlet pipe 50. The connecting pipe 44 and the outlet pipe 50 are connected as a whole.
[0045] Step S4 specifically includes the following steps:
[0046] S42, placing the dispersed phase 1 in the first syringe 32, and placing the dispersed phase 2 in two second syringes 34 respectively;
[0047] S44. The dispersed phase 1 in the first syringe 32 is injected into the first inlet pipe 42 through the first injection pump, and the dispersed phase 2 in the two second syringes 34 are respectively injected into the two second inlet pipes 43 through the two second injection pumps. In the connecting pipe 44, the dispersed phase 1 is injected into the dispersed phase 2 in the two second inlet pipes 43 and is sheared into organoid precursor droplets 100 of uniform size. The sheared organoid precursor droplets 100 can enter the outlet pipe 50 in sequence and can be matured into organoid spheres 102 in the outlet pipe 50.
[0048] S6. The first organoid sphere 102 in the organoid droplet maturation zone is transported to the print head 62 of the 3D printing system through the transport pipe 52 of the 3D printing system by the transport pump of the microfluidic system.
[0049] In step S6, the 3D printing system is a 3D bio-printer.
[0050] S8. Detect whether there is an organoid sphere 102 at the print head 62 through the organoid recognition detection unit 64 of the 3D printing system. When the organoid recognition detection unit 64 detects that there is an organoid sphere 102 at the print head 62, the organoid recognition detection unit 64 outputs an organoid signal to the control unit of the 3D printing system.
[0051] In step S8, the organoid identification and detection unit 64 is opposite to the printing nozzle 62, such as Figure 2 As shown, the organoid sphere 102 is detected by the organoid identification and detection unit 64, such as color, geometric outline, fluorescent signal, etc., so that the presence of the organoid sphere 102 at the printing nozzle 62 can be detected. The organoid identification and detection unit 64 is, for example, a CCD camera.
[0052] S10, controlling the action of the driving unit of the 3D printing system through the control unit, driving the print nozzle 62 to move downward to the bottom of the vascular bed culture chamber 202 of the first vascular bed microfluidic chip 200, and staying there for 1-2 seconds, preferably 1 second. Under the adhesion effect between the first organoid sphere 102 and the bottom of the vascular bed culture chamber 202, the first organoid sphere 102 will be adsorbed on the bottom of the vascular bed culture chamber 202 of the first vascular bed microfluidic chip 200, thereby printing the first organoid sphere 102 into the vascular bed culture chamber 202 of the first vascular bed microfluidic chip 200. After completion, the print nozzle 62 is driven by the driving unit to move upward to return to the initial position, which is the relative position of the print nozzle 62 and the organoid identification and detection unit 64.
[0053] In this embodiment, the moving speed range of the print head 62 is generally 0-100 mm / s (millimeter / second) on the XY axis and 0-80 mm / s on the Z axis. The moving speed and acceleration of the print head 62 can be adjusted in real time and flexibly according to the printing speed and accuracy requirements.
[0054] In step S10 , the vascular bed culture chamber 202 has an opening, and the print head 62 moves to the bottom of the vascular bed culture chamber 202 through the opening.
[0055] S12. Print the remaining organoid spheres 102 in the organoid droplet maturation zone into the remaining vascular bed culture chambers 202 of the vascular bed microfluidic chip 200 in sequence according to the methods of steps S6 to S10, thereby completing high-throughput printing of organoids.
[0056] S14, culturing the organoid spheroids 102 at the bottom of the vascular bed culture chambers 202 of all vascular bed microfluidic chips 200, that is, the organoid spheroids 102 are stably present at the bottom of the vascular bed culture chambers 202 for culturing, such as Fig. 9 shown.
[0057] In step S14 , when the organoid spheroid 102 is cultured, the dispersed phase 2 will evaporate by itself because the material of the dispersed phase 2 includes fluorinated oil.
[0058] The high-throughput organoid culture method provided by the present invention completes the preparation of organoids through a microfluidic system, and by combining the microfluidic system with a 3D printing system, the organoids can be easily, quickly and accurately transferred to the bottom of the vascular bed culture chamber 202 of the vascular bed microfluidic chip 200 for culture by printing. The process has good controllability, saves time and effort, has good stability, and the organoids are uniform in size, which ensures the consistency of the organoids and reduces the differences between individual organoids. At the same time, printing the organoids into the vascular bed culture chamber 202 of the vascular bed microfluidic chip 200 for culture can simulate the microenvironment in the human body, and a complete vascular network can be constructed inside the organoid within a short time (such as 3-5 days), shortening the growth and development time of the organoids and improving the efficiency and accuracy of drug screening.
[0059] The above is a specific description of the preferred implementation of the present invention, but the invention is not limited to the embodiments. Those skilled in the art can make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.
Claims
1. A high-throughput organoid culture method, characterized in that: The following steps are involved: S2, encapsulating the cells in dispersed phase 1; S4, respectively injecting dispersed phase 1 and dispersed phase 2 encapsulating cells into the organoid precursor preparation area of the microfluidic system, dispersed phase 1 is sheared into organoid precursor droplets of uniform size by dispersed phase 2, and the sheared organoid precursor droplets can sequentially enter the organoid droplet maturation area of the microfluidic system and can be matured into organoid spheres in the organoid droplet maturation area; S6, using a delivery pump of the microfluidic system to deliver the first organoid sphere in the organoid droplet maturation zone to the printing nozzle of the 3D printing system through a delivery pipeline of the 3D printing system; S8, detecting whether there is an organoid sphere at the printing nozzle by the organoid recognition and detection unit of the 3D printing system, and when the organoid recognition and detection unit detects that there is an organoid sphere at the printing nozzle, the organoid recognition and detection unit outputs an organoid signal to the control unit of the 3D printing system; S10, controlling the action of the driving unit of the 3D printing system through the control unit, driving the printing nozzle to move downward to the bottom of the vascular bed culture chamber of the first vascular bed microfluidic chip through the driving unit, and after staying for 1-2 seconds, the first organoid sphere will be adsorbed on the bottom of the vascular bed culture chamber of the first vascular bed microfluidic chip, so that the first organoid sphere is printed into the vascular bed culture chamber of the first vascular bed microfluidic chip. After completion, the printing nozzle is driven by the driving unit to return to the initial position; S12, printing the remaining organoid spheres in the organoid droplet maturation area into the vascular bed culture chamber of the remaining vascular bed microfluidic chip in sequence according to the methods of step S6 to step S10; S14, culturing the organoid spheroids in the vascular bed culture chambers of all vascular bed microfluidic chips; In step S4, the method of maturing the organoid precursor droplets into organoid spheres includes enzymatic crosslinking, ionic crosslinking or photocrosslinking; The material of the dispersed phase 1 includes matrix gel, collagen or gelatin, and the material of the dispersed phase 2 includes fluorinated oil or vegetable oil.
2. The high-throughput organoid culture method according to claim 1, characterized in that: The organoid precursor preparation area is a Y-shaped structure, and the Y-shaped structure includes a first inlet pipe, a second inlet pipe and a connecting pipe, the first end of the first inlet pipe and the first end of the second inlet pipe are respectively connected to the first syringe and the second syringe, the first syringe is connected to the first injection pump, the second syringe is connected to the second injection pump, the second end of the first inlet pipe and the second end of the second inlet pipe are both connected to the first end of the connecting pipe, and the organoid droplet maturation area is an outlet pipe, and the second end of the connecting pipe is connected to the outlet pipe; The step S4 comprises the following steps: S42, placing dispersed phase 1 and dispersed phase 2 in a first syringe and a second syringe respectively; S44. The dispersed phase 1 in the first syringe is injected into the first inlet pipe through the first injection pump, and the dispersed phase 2 in the second syringe is injected into the second inlet pipe through the second injection pump. In the connecting pipe, the dispersed phase 1 is sheared into organoid precursor droplets of uniform size by the dispersed phase 2. The sheared organoid precursor droplets can enter the outlet pipe in turn and can be matured into organoid spheres in the outlet pipe.
3. The high-throughput organoid culture method according to claim 1, characterized in that: The organoid precursor preparation area is a T-shaped structure, and the T-shaped structure includes a first inlet pipe, a second inlet pipe and a connecting pipe, the first end of the first inlet pipe and the first end of the second inlet pipe are respectively connected to the first syringe and the second syringe, the first syringe is connected to a first injection pump, the second syringe is connected to a second injection pump, the second end of the first inlet pipe and the second end of the second inlet pipe are respectively connected to the first end of the connecting pipe, and the organoid droplet maturation area is an outlet pipe, and the second end of the connecting pipe is connected to the outlet pipe; The step S4 comprises the following steps: S42, placing dispersed phase 1 and dispersed phase 2 in a first syringe and a second syringe respectively; S44. The dispersed phase 1 in the first syringe is injected into the first inlet pipe through the first injection pump, and the dispersed phase 2 in the second syringe is injected into the second inlet pipe through the second injection pump. In the connecting pipe, the dispersed phase 1 is sheared into organoid precursor droplets of uniform size by the dispersed phase 2. The sheared organoid precursor droplets can enter the outlet pipe in turn and can be matured into organoid spheres in the outlet pipe.
4. The high-throughput organoid culture method according to claim 1, characterized in that: The organoid precursor preparation area is a cross-shaped structure, which includes a first inlet pipe, two second inlet pipes arranged opposite to each other and a connecting pipe, the first inlet pipe and the connecting pipe are arranged opposite to each other, the first end of the first inlet pipe is connected to the first syringe, the second end of the first inlet pipe is connected to the first end of the connecting pipe, the first syringe is connected to a first injection pump, the first ends of the two second inlet pipes are respectively connected to the two second syringes, the second ends of the two second inlet pipes are respectively connected to the first end of the connecting pipe, the two second syringes are respectively connected to two second injection pumps, and the organoid droplet maturation area is an outlet pipe, and the second end of the connecting pipe is connected to the outlet pipe; The step S4 comprises the following steps: S42, placing dispersed phase 1 in a first syringe, and placing dispersed phase 2 in two second syringes respectively; S44. The dispersed phase 1 in the first syringe is injected into the first inlet pipe through the first injection pump, and the dispersed phase 2 in the two second syringes are respectively injected into the two second inlet pipes through the two second injection pumps. In the connecting pipe, the dispersed phase 1 injected into the dispersed phase 2 in the two second inlet pipes is sheared into organoid precursor droplets of uniform size. The sheared organoid precursor droplets can enter the outlet pipe in sequence and can be matured into organoid spheres in the outlet pipe.
5. The high-throughput organoid culture method according to claim 1, characterized in that: In step S2, the cells are primary tumor tissue cells, stem cells or cell lines.
6. The high-throughput organoid culture method according to claim 1, characterized in that: In step S8, the organoid recognition and detection unit is opposite to the printing nozzle, and the organoid recognition and detection unit detects the color, geometric outline or fluorescent signal of the organoid sphere, so that the presence of the organoid sphere at the printing nozzle can be detected.