A cell trap generation module for standardized and controllable regeneration of organoids

Abstract

The present invention relates to the field of biological tissue engineering technology, and discloses a cell trap generation module for standardized and controllable regeneration of organoids, including a dehydration module, a first cell carrier, a second cell carrier, a first film-covered support, and a second film-covered support; the first cell carrier is arranged in the first film-covered support, and a concave hole array is arranged on the first cell carrier, and cells are arranged in the concave hole array; the second cell carrier is arranged in the second film-covered support; a dehydration channel is arranged in the dehydration module, and the first cell carrier is at least partially exposed to the dehydration channel for dehydration, the second film-covered support is arranged outside the first cell carrier, and the second cell carrier is opposite to the concave hole array of the first cell carrier, and the second cell carrier enters the concave hole array and wraps the cells to form a cell trap array. The cell trap generation module for standardized and controllable regeneration of organoids of the present invention can simplify the manufacturing process of organoids and improve the controllability and consistency of the organoid regeneration process.

Description

Technical Field

[0001] The present invention relates to the technical field of biological tissue engineering, and in particular to a cell trap generation module for standardized and controllable regeneration of organoids. Background Art

[0002] Organoids are tissue models obtained by culturing adult stem cells or pluripotent stem cells in vitro that can simulate the characteristics of corresponding organs in vivo. They have broad prospects in basic research, disease modeling, drug development, precision treatment and other fields.

[0003] High-throughput culture of organoids can be achieved through culture systems such as microwell arrays and microfluidic droplets. However, the current manufacturing process of organoids relies on complex manual operations by experimenters, which makes it difficult to automate and mass-produce them, and the controllability and consistency of the organoid regeneration process are difficult to guarantee. Since the manufacturing process requires fine micro-scale operations, the time cost required to train related personnel is also very high.

[0004] Therefore, it is necessary to develop a cell trap generation module for standardized and controllable regeneration of organoids to simplify the organoid manufacturing process and improve the controllability and consistency of the organoid regeneration process. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a cell trap generation module for standardized and controllable regeneration of organoids, so as to simplify the manufacturing process of organoids and improve the controllability and consistency of the organoid regeneration process.

[0006] In order to solve the above technical problems, the present invention provides a cell trap generation module for standardized and controllable regeneration of organoids, comprising a dehydration module, a first cell carrier, a second cell carrier, a first coated stent and a second coated stent;

[0007] The first cell carrier is disposed in the first membrane-covered stent, the first cell carrier is provided with a concave hole array, and cells are arranged in the concave hole array;

[0008] The second cell carrier is disposed in the second coated stent;

[0009] A dehydration channel is provided in the dehydration module, the first film-covered support is detachably connected to the dehydration module, the first cell carrier is at least partially exposed in the dehydration channel for dehydration, the second film-covered support is arranged outside the first cell carrier, and the second cell carrier is opposite to the concave hole array of the first cell carrier, the second cell carrier enters the concave hole array and wraps the cells to form a cell trap array.

[0010] As an improvement of the above solution, the surface of the first cell carrier opposite to the concave hole array is exposed in the dehydration channel.

[0011] As an improvement of the above solution, the first cell carrier and the first membrane-covered stent isolate the second cell carrier from the dehydration channel.

[0012] As an improvement of the above solution, the top surface of the second cell carrier is covered by the second membrane-covered stent.

[0013] As an improvement of the above solution, the first coated stent includes a stent body and a first supporting boss. A through hole is provided in the stent body. The first supporting boss is arranged along the circumference of the through hole. The first supporting boss is used to support the first cell carrier.

[0014] As an improvement of the above solution, the first stent graft further includes a support arm disposed on the outer periphery of the stent body;

[0015] The first end of the dehydration module is provided with a mounting groove which is arranged in rotational symmetry. The mounting groove extends spirally and is arranged corresponding to the support arm. The support arm is arranged on the outer periphery of the bracket body along the spiral direction of the mounting groove. The support arm is screwed into the mounting groove, and the bracket body extends into the dehydration channel.

[0016] As an improvement of the above scheme, it also includes an array mold and a support assembly, the first film-covered bracket is detachably connected to the dehydration module, the first film-covered bracket and the support assembly cooperate to form a receiving cavity, the receiving cavity is provided with an inert gel, the inert gel is pressed by the array mold to form the concave hole array, forming the first cell carrier, and the first cell carrier can be pulled out of the support assembly along with the first film-covered bracket.

[0017] As an improvement of the above-mentioned scheme, the support assembly includes a culture dish and a support, the support is arranged in the culture dish, the support includes a base and a second support boss formed on the base, the second support boss is adapted to the through hole, and the second support boss extends from the bottom of the through hole to form the accommodating cavity.

[0018] As an improvement of the above scheme, it also includes a dehydration platform, which is provided with a heat source cavity, an installation cavity and a dehydration hole. The installation cavity is used to accommodate the dehydration module, and the dehydration hole is used to connect the heat source cavity with the dehydration channel so that the heat source of the heat source cavity flows to the dehydration channel.

[0019] As an improvement of the above-mentioned scheme, the side wall of the installation cavity is provided with a first limiting groove and a second limiting groove, the second limiting groove is connected to the bottom end of the first limiting groove, and the second limiting groove is extended from the first limiting groove to the circumference of the installation cavity, and the second end of the dehydration module is provided with a mounting protrusion protruding toward the outer wall, and the mounting protrusion is inserted from the top end of the first limiting groove and screwed into the second limiting groove.

[0020] The implementation of the present invention has the following beneficial effects:

[0021] The present invention discloses a cell trap generation module for standardized and controllable regeneration of organoids. In the process of arranging a concave hole array on a first cell carrier, arranging cells in the concave hole array, and exposing the first cell carrier at least partially to the dehydration channel for dehydration, the concave hole array structure on the first cell carrier enables the second cell carrier opposite to the first cell carrier to enter the concave hole array and wrap the cells therein, and also to achieve dehydration. The dehydration of the second cell carrier forms an entangled state on the side opposite to the concave hole array. The part forming the entangled state through dehydration increases the physical strength, changes from a very soft state to a harder solid state, and is easier to separate from the first cell carrier, so that the cells remain in the second cell carrier to form a cell trap array.

[0022] Since the first cell carrier and the second cell carrier are easier to separate after dehydration, the operation of the organoid manufacturing process is simplified;

[0023] Since the first coated support is detachably connected to the dehydration module, and the first cell carrier is at least partially exposed to the dehydration channel when installed on the dehydration module, the first coated support can be directly used as a dehydration carrier and can be easily moved. The concave hole array can be directly formed on the first coated support, which simplifies the production and transfer operations of the first cell carrier;

[0024] The first cell carrier has a concave hole array for implanting cells, which provides a controllable growth space for the implanted cells, prevents the cells from expanding and growing outward in the horizontal direction, and can improve the consistency of the cell-like regeneration process;

[0025] The second cell carrier encircles and wraps the cells in the concave hole array to form a cell well array. Each cell well on the second cell carrier is a multi-cell system. Each cell in it is mutually constrained with other cells. The time and spatial characteristics of the activation of cell signals will gradually stabilize, and it has a certain resistance to external disturbances, which can help improve the controllability and consistency of the organoid regeneration process. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a schematic structural diagram of an embodiment of a cell trap generation module for standardized and controllable regeneration of organoids according to the present invention;

[0027] Figure 2 is a schematic diagram of the assembly structure of a first membrane-covered support with a first cell carrier and a dehydration module;

[0028] Figure 3 yes Figure 2 Side view of

[0029] Figure 4 yes Figure 3 Schematic diagram of CC cross-section structure;

[0030] Figure 5 yes Figure 1 It is a structural schematic diagram of an array mold, a support assembly and a first coated stent in another embodiment of a cell trap generation module for standardized and controllable regeneration of organoids of the present invention;

[0031] Figure 6 is a schematic structural diagram of a first stent graft;

[0032] Figure 7 It is a structural diagram of the support;

[0033] Figure 8 It is a schematic diagram of the assembly structure of the dehydration platform and the dehydration module;

[0034] Fig. 9 yes Figure 8 A top view of

[0035] Fig.10 yes Fig. 9 Schematic diagram of the DD cross-section structure.

[0036] Fig.11 It is a schematic diagram of the structure of a cell trap array automated manufacturing system based on human-machine interaction of a cell trap generation module;

[0037] Fig.12 yes Fig.11 Structural diagram of the workbench, robotic arm, and industrial camera; DETAILED DESCRIPTION

[0038] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to the accompanying drawings.

[0039] like Figures 1 to 4As shown, the present invention discloses an embodiment of a cell trap generation module for standardized and controllable regeneration of organoids, comprising a dehydration module 14, a first cell carrier b, a second cell carrier c, a first film-covered support 12 and a second film-covered support 15; the first cell carrier b is arranged in the first film-covered support 12, and the first cell carrier b is provided with a concave hole array b1, and cells are arranged in the concave hole array b1; the second cell carrier c is arranged in the second film-covered support 15; a dehydration channel 141 is provided in the dehydration module 14, the first film-covered support 12 is detachably connected to the dehydration module 14, the first cell carrier b is at least partially exposed in the dehydration channel 141 for dehydration, the second film-covered support 15 is arranged outside the first cell carrier b, and the second cell carrier c is opposite to the concave hole array b1 of the first cell carrier b, the second cell carrier c enters the concave hole array b1 and wraps the cells to form a cell trap array.

[0040] In this embodiment, the first cell carrier b is made of an inert gel a, preferably agarose gel. The second cell carrier c is a biological hydrogel such as matrigel, collagen, fibrinogen-thrombin, etc., and the present embodiment is preferably a matrigel commonly used in organoid cultivation. The matrigel wraps the cultured cells so that the environment around the cells has better consistency, and there will be no polarity reversal due to the polarity difference of the micro-surroundings, and it can simulate the in vivo environment as much as possible. The second film-covered support 15 is preferably the first culture dish. The top surface of the second cell carrier c is covered by the first culture dish, and the first cell carrier b and the first film-covered support 12 isolate the second cell carrier c from the dehydration channel 141. When the first culture dish with matrigel inside is covered on the agarose gel with the concave hole array b1, the matrigel is attached to the first culture dish.

[0041] Agarose gel cannot form a covalent bond with all current biohydrogels due to its inert chemical properties. The present invention has found in the research process that many biohydrogels used to make organoids, including matrix gel, can form an entangled state by dehydration, and can provide a gel with physical strength beyond cross-linking in a short time. Therefore, after the second cell carrier c contacts the first cell carrier b, no chemical bond is formed. In addition, the second cell carrier c in the second coated support 15 of this embodiment is arranged relative to the concave hole array b1. During the dehydration process of the first cell carrier b, the first cell carrier b and the first coated support 12 isolate the second cell carrier c from the dehydration channel 141. Although the second cell carrier c is not directly exposed to the dehydration channel 141, the concave hole array b1 structure on the first cell carrier b enables the second cell carrier c to be dehydrated, and the second cell carrier c will form an entangled state on the side opposite to the concave hole array b1. When the second cell carrier c of this embodiment enters the concave hole array b1 and wraps the cells, the matrix gel in the entangled state after dehydration will increase its physical strength, changing from a very soft state to a harder solid state, making it easier to separate from the first cell carrier b. When the side of the second cell carrier c opposite to the concave hole array b1 is dehydrated and entangled, the side of the second cell carrier c in contact with the first culture dish still maintains its original state, that is, the top of the second cell carrier c is adsorbed on the first culture dish, and the matrix gel and agarose gel can be completely detached by gently pulling with the hand, so that the matrix gel wrapped with cells remains on the first culture dish, and the cell trap array is completed.

[0042] The first cell carrier b has a concave hole array b1 for implanting cells, which provides a controllable growth space for the implanted cells and prevents the cells from expanding and growing outward in the horizontal direction, thereby improving the consistency of the cell-like regeneration process; the second cell carrier c encloses and wraps the cells in the concave hole array b1 to form a cell well array, and each cell well on the second cell carrier c is a multi-cell system, in which each cell is mutually constrained with other cells, and the time and space characteristics of the activation of its cell signal will gradually stabilize, and it has a certain resistance to external disturbances.

[0043] The first membrane-covered support 12 of the present embodiment specifically includes a support body 121 and a first support boss 122, the support body 121 is provided with a through hole 123, and the first support boss 122 is arranged along the circumference of the through hole 123. The first membrane-covered support 12 is arranged in a support assembly. The support assembly is provided with a second support boss 161, the second support boss 161 is adapted to the through hole 123, the second support boss 161 extends from the bottom of the through hole 123, and forms a receiving cavity with the through hole 123, and the inert gel a in the receiving cavity contacts the first support boss 122.

[0044] In this embodiment, after agarose is poured into the accommodating cavity, the agarose forms an agarose gel, and the agarose gel is coated on the first supporting boss 122, so that the first supporting boss 122 can be embedded in the agarose gel to form a stable support for it.

[0045] The bottom of the array mold 11 is provided with a protrusion array corresponding to the concave hole array b1. The array mold 11 can directly press the concave hole array b1 on the top surface of the inert gel a, or press on the top of the poured agarose. After solidification, the array mold 11 is removed to obtain an agarose gel with a concave hole array b1. When the array mold 11 presses the surface of the agarose gel, the protrusion array is opposite to the second supporting boss 161 and staggered with the first supporting boss 122. After the pressing is completed, the array mold 11 is pulled out to form a concave hole array b1 in the middle area of ​​the agarose gel surface. The thickness of the agarose gel in the middle area is uniform, which is conducive to improving the consistency of the cell trap. The agarose gel forming the concave hole array b1 is the first cell carrier b. By moving the first film-covered support 12, the first cell carrier b can be transferred to the cell inoculation area B with the first film-covered support 12 for cell inoculation.

[0046] The protrusions of the protrusion array on the array mold 11 are conical, and the concave hole array b1 structure formed by the array mold 11 with this structure makes it easier for the second cell carrier c to separate the encapsulated cells from the first cell carrier b.

[0047] In order to facilitate the movement of the first coated stent 12 and the first cell carrier b thereon, the first coated stent 12 of the present embodiment is further provided with support arms 124 , which are arranged in an array along the periphery of the stent body 121 .

[0048] The support assembly of this embodiment specifically includes a second culture dish 17 and a support 16 disposed in the second culture dish 17. Among them, the support 16 includes a base 162, a limiting boss 163, and the above-mentioned second support boss 161, the second support boss 161 and the limiting boss 163 are both formed on the top of the base 162, the limiting boss 163 is arranged in an array around the periphery of the second support boss 161, the support body 121 is arranged between the limiting boss 163 and the second support boss 161, and the support arm 124 is arranged between two adjacent limiting bosses 163. The second culture dish 17 can collect materials overflowed during the pouring of agarose. The support 16 structure enables the first film-covered support 12 to be placed in a directional manner, which facilitates the rapid positioning and pressing of the array mold 11 and facilitates the molding of the concave hole array b1. In addition, by designing the support 16 with the second support boss 161 of different heights, the height of the second support boss 161 extending into the through hole 123 is changed, thereby changing the thickness of the agarose gel to meet different organoid cultivation requirements. The support 16 cooperates with the first coated support 12 to form a structure of a receiving cavity for accommodating agarose, so that agarose gels of different thicknesses can be obtained without having to design different models of the first coated support 12.

[0049] The first end of the dehydration module 14 of this embodiment is provided with a mounting groove 142 which is arranged in rotational symmetry, and the second end of the dehydration module 14 is connected to the dehydration platform 13. Among them, the mounting groove 142 extends spirally and is arranged corresponding to the support arm 124, and the support arm 124 is arranged along the spiral direction of the mounting groove 142 on the periphery of the support body 121. When cell inoculation or dehydration is performed, the concave hole array b1 is kept upward, the support arm 124 of the first film support 12 is screwed into the mounting groove 142, and the support body 121 extends into the dehydration channel 141. At this time, the through hole 123 is coaxial with the dehydration channel 141, and the surface of the first cell carrier b opposite to the concave hole array b1 is exposed in the dehydration channel 141, so that the first cell carrier b is quickly dehydrated, and the second cell carrier c is dehydrated through the concave hole array; the top surface of the second cell carrier c is covered by the second film support 15, and a large adsorption area is maintained between the second cell carrier c and the second film support 15, which is convenient for the subsequent separation of the second cell carrier c from the first cell carrier b.

[0050] Among them, a sink 144 is provided in the dehydration channel 141, and the first coated support 12 is spirally connected to the dehydration module 14. After the support body 121 extends into the dehydration channel 141, the support body 121 will abut against the sink 144 to close the dehydration channel 141, thereby isolating the second cell carrier c from the dehydration channel 141. The first coated support 12 is not only easy to move and position, but also plays a role in controlling the flow of heat source.

[0051] More than two dehydration channels 141 may be provided on the dehydration module 14 , and the top of each dehydration channel 141 is connected to the first membrane-covered support 12 to achieve higher throughput of cell inoculation.

[0052] The first cell carrier b of this embodiment is supported by the first film support 12. The support arm 124 structure of the first film support 12 cooperates with the spiral mounting groove 142 of the dehydration module 14 to facilitate the disassembly and assembly between the first film support 12 and the dehydration module 14. The through hole 123 structure of the first film support 12 can promote the dehydration of the first cell carrier b and the second cell carrier c.

[0053] The dehydration platform 13 is provided with an installation cavity 132 and a dehydration hole 133. The installation cavity 132 is used to accommodate the dehydration module 14. The dehydration hole 133 is used to connect the heat source cavity 131 with the dehydration channel 141, so that the hot air of the heat source cavity 131 flows to the dehydration channel 141. The side wall of the installation cavity 132 is provided with a first limiting groove 134 and a second limiting groove 135. The second limiting groove 135 is connected to the bottom end of the first limiting groove 134, and the second limiting groove 135 is extended from the first limiting groove 134 to the circumferential direction of the installation cavity 132. The second end of the dehydration module 14 is provided with a mounting protrusion 143 protruding toward the outer wall. The mounting protrusion 143 is inserted from the top of the first limiting groove 134 and screwed into the second limiting groove 135.

[0054] See also Figures 1 to 12The present invention also discloses an embodiment of a human-machine interactive cell trap array automated manufacturing system based on a cell trap generation module, comprising a workbench 1, a robotic arm 2, an industrial camera 3 and a control module 4. The workbench 1 is provided with an array production area A and a cell inoculation area B. The array production area A is provided with an array mold 11, a support assembly, and a first film-covered support 12. The first film-covered support 12 cooperates with the support assembly to form a receiving cavity. An inert gel a is provided in the receiving cavity. The inert gel a is pressed by the array mold 11 on the surface to form a concave hole array b1 to form a first cell carrier b. The first cell carrier b can be removed from the support assembly with the first film-covered support 12; the cell inoculation area B is provided with a dehydration platform 13 and a dehydration module 14. The dehydration module 14 is provided with a dehydration channel 141. The dehydration platform 13 is provided with a heat source cavity 131 for providing hot air to the dehydration channel 141. The dehydration channel 141 is connected to the heat source cavity 131. The first film-covered support 12 carrying the first cell carrier b is detachably connected to the dehydration module 14. The industrial camera 3 is arranged above the cell inoculation area B, and is used to visually image the concave hole array b1 of the cell inoculation area B to obtain the position image of the concave hole array b1. The control module 4 is electrically connected to the mechanical arm 2 and the industrial camera 3. The driving end of the mechanical arm 2 is provided with a pipette 21. The control module 4 controls the mechanical arm 2 to drive the pipette 21 to implant cells into the concave hole array b1 corresponding to the position image of the concave hole array b1 according to the position image of the concave hole array b1 fed back by the industrial camera 3. After the cell inoculation and cultivation are completed, a second film-covered support 15 is arranged outside the first cell carrier b, and a second cell carrier c arranged opposite to the concave hole array b1 is arranged inside the second film-covered support 15. The first cell carrier b is at least partially exposed to the dehydration channel 14 for dehydration, and the second cell carrier c enters the concave hole array b1 and wraps the cells to form a cell trap array.

[0055] In this embodiment, the workbench 1 is modularly partitioned and arranged to set an array production area A and a cell inoculation area B. In the array production area A, the inert gel a and the concave hole array b1 on its top surface are formed by the cooperation of the array mold 11, the support assembly, and the first coated support 12 to obtain a first cell carrier b that can be separated from the support assembly with the first coated support 12; in the cell inoculation area B, the first coated support 12 carrying the first cell carrier b is detachably connected to the dehydration module 14, and the concave hole array b1 is visually imaged by the industrial camera 3 above the cell inoculation area B to obtain a position image of the concave hole array b1. The control module 4 controls the robot arm 2 to drive the pipette 21 to implant cells into the concave hole array b1 according to the position image of the concave hole array b1 fed back by the industrial camera 3; after the first cell carrier b implanted with cells is detached from the dehydration module 14 with the first coated support 12, it can be placed in a culture environment to culture the cells, and the concave hole array b1 provides a physical boundary for the growth of cell spheres or organoids, so that cell spheres or organoids with controllable types, quantities, and densities are obtained on the first cell carrier b; after the cells are in the first After the cell carrier b completes self-assembly, the first film support 12 carrying the first cell carrier b is detachably connected to the dehydration module 14, so that the surface of the first cell carrier b opposite to the concave hole array b1 is exposed in the dehydration channel 141, and a second film support 15 is arranged outside the first cell carrier b, so that the second cell carrier c in the second film support 15 is arranged opposite to the concave hole array b1, and the second cell carrier c will enter the concave hole array b1 and wrap the cells. At the same time, under the action of the hot air in the dehydration channel 141, the first cell carrier b is quickly dehydrated, and the second cell carrier c is easily separated from the first cell carrier b, so that the cells remain in the second cell carrier c to form a cell trap array; this embodiment provides a human-machine interactive robot arm 2, and cooperates with the industrial camera 3 to assist the operator's fine movements. At the same time, the high-throughput manufacturing process of organoids is simplified through the structural coordination between the dehydration platform 13, the dehydration module 14, the first film support 12, the array mold 11, the support assembly, and the second film support 15 on the workbench 1, thereby improving the controllability and consistency of the organoid regeneration process.

[0056] The first coated support 12 can be used in the array fabrication area A to form the concave hole array b1, and can also be directly moved to the cell inoculation area B as a dehydrated carrier, thereby helping to simplify the fabrication and transfer operations of the first cell carrier b.

[0057] The heat source in the heat source cavity 131 is provided by the air compressor 5 and the gas heater 6, and the air compressor 5, the gas heater 6, and the heat source cavity 131 are connected in sequence. The air compressor 5 draws air from the environment and pressurizes it to a certain pressure. The pressure air is heated to a certain temperature by the gas heater 6 and then input into the heat source cavity 131 of the dehydration platform 13 to provide hot convection air for the dehydration module 14. The gas heater 6 includes a refrigeration module and a heating module to achieve an adjustable temperature effect. The dehydration channel 141 of the dehydration module 14 is connected with the refrigeration module and the heating module to provide the dehydration channel 141 with adjustable temperature hot convection air. The dehydration platform 13 is provided with an air inlet 136 and an exhaust hole 137 connected with the heat source cavity 131. The pressure air is heated to a certain temperature by the gas heater 6 and then input into the heat source cavity 131 through the air inlet 136.

[0058] A mounting plate 18 for mounting the dehydration platform 13 is provided on the workbench 1, and an assembly groove 181 and a positioning groove 182 are provided on the mounting plate 18. The assembly groove 181 is adapted to the dehydration platform 13, and the assembly groove 181 is communicated with the positioning groove 182. An elastic positioning piece (not shown in the figure) is provided in the positioning groove 182, which is used to elastically abut against the side wall of the dehydration platform 13 in the assembly groove 181 to realize rapid positioning and installation of the dehydration platform 13.

[0059] The dehydration module 14 on the dehydration platform 13 is used not only to provide a dehydration channel 141 during the dehydration process, but also to fix the first membrane-covered support 12 during cell inoculation. In order to achieve higher throughput cell inoculation, this embodiment arranges multiple dehydration modules 14 in an array on the dehydration platform 13 in the cell inoculation area B, so that multiple first membrane-covered supports 12 with first cell carriers b are installed in the cell inoculation area B. Each dehydration module 14 can be removed from the dehydration platform 13 individually, which is more convenient for testing.

[0060] In addition, the cell inoculation area B is also provided with an XY electric translation stage 19, and the dehydration platform 13 is provided on the top of the XY electric translation stage 19, which is used to drive the dehydration platform 13 to move in the horizontal and vertical directions, so that the first cell carrier b on different dehydration modules 14 falls into the visual range of the industrial camera 3, and the mechanical arm 2 drives the liquid transfer gun 21 to implant cells into the concave hole array b1 on the first cell carrier b that falls into the visual range of the industrial camera 3. The cell types in the liquid transfer gun 21 can be set as needed, and the number of cells implanted in the concave hole array b1 can be controlled by the liquid transfer gun 21. By controlling the implantation of cells into concave holes at different intervals through the mechanical arm 2 or the XY electric translation stage 19, the cell density distribution of the concave hole array b1 can be controlled.

[0061] The control module 4 of this embodiment uses a computer as the control center of the entire system to process information from various parts and issue control instructions.

[0062] The workbench 1 of this embodiment is modularly distributed. In addition to the above-mentioned array production area A and cell inoculation area B, a robot arm 2 module area and a microscopic vision area are also provided. The robot arm 2 is arranged in the robot arm 2 module area of ​​the workbench 1, and the industrial camera 3 is arranged in the microscopic vision area of ​​the workbench 1. The modules are connected by a bottom plate to form an integral structure.

[0063] The driving end of the mechanical arm 2 is provided with an electric clamp 22, and the liquid transfer gun 21 is clamped by the electric clamp 22. The mechanical arm 2 can realize multi-degree-of-freedom, high-speed, and high-precision movement. The mechanical arm 2 and the electric clamp 22 can cooperate to grasp objects in a large range, at high speed, and with high precision, and move the objects to the corresponding position to realize human-computer interaction. The electric clamp 22 can clamp the liquid transfer gun 21 to realize fast, accurate, and stable cell inoculation.

[0064] The microscopic visual area includes an industrial camera 3, a lens, a light source and a fixed bracket. The lens and the light source are both arranged on the industrial camera 3. The light source fill light and the lens focus can help obtain a clearer image. The industrial camera 3 is fixed on the workbench 1 through a fixed bracket. The position image of the concave hole array b1 on the first cell carrier b is captured by the industrial camera 3. After computer image processing, the robot arm 2 and the XY electric translation stage 19 are controlled to move accordingly. Through the translation of the XY electric translation stage 19 along the XY directions, the industrial camera 3 can observe the conditions of the concave hole arrays b1 on different first cell carriers b under the microscope.

[0065] In addition to fixing the robot arm 2 and the industrial camera 3 on the workbench 1, the industrial camera 3 can also be fixed on the driving end of the robot arm 2 to realize the automation of recognition and operation through computer vision technology.

[0066] In order to ensure a clean experimental environment, a biosafety cabinet 7 is also provided on the workbench 1 in this embodiment. The biosafety cabinet 7 is provided with a working area, a ventilation system, a filtration system, a UV lamp, a protective barrier, etc. The workbench 1 is located in the working area of ​​the biosafety cabinet 7, so that the organoid array can be manufactured in the working area of ​​the safety cabinet to minimize contamination.

[0067] In addition, according to specific needs, equipment modules such as a vibrator, a centrifuge, and a PCR instrument can also be set on the workbench 1.

[0068] The working process of the present invention is as follows: the support 16 is placed in the second culture dish 17, and then the first film support 12 is placed on the corresponding support 16, agarose is poured into the first film support 12, and then the array mold 11 is pressed on the first film support 12. After the agarose solidifies, the array mold 11 is pulled out to obtain the first cell carrier b solidified on the first film support 12. The first film support 12 is placed on the dehydration module 14. Under the visual navigation of the industrial camera 3, the experimenter controls the mechanical arm 2 and the electric gripper 22 to clamp the liquid transfer gun 21 and move it to the top of the dehydration platform 13. By controlling the XY electric translation stage 19, the dehydration module 14 moves accurately under the liquid transfer gun 21, and cells are inoculated in the concave hole array b1 of the first cell carrier b in turn. After the cells are inoculated, the first film support 12 is removed and placed in the second culture dish 17 or a six-well plate for culture, so that a cell array with controllable cell types, quantities and densities is quickly manufactured. After the cells complete self-assembly in the first cell carrier b, the first membrane-covered support 12 is taken out from the second culture dish 17 or the six-well plate, placed on the dehydration module 14, and the second culture dish 17 with the base of the base gel evenly coated on the first cell carrier b is covered. At this time, the base gel will enter the concave hole array b1 and wrap the cultivated cell spheres or organoids. At the same time, the constant temperature air generated by the air compressor 5 and the gas heater 6 convects with the first cell carrier b, so that the first cell carrier b and the second cell carrier c on the side opposite to the concave hole array b1 are quickly taken away. The second cell carrier c made of base gel forms an entangled state during the dehydration process, and can be physically separated from the first cell carrier b relatively easily. Therefore, the second culture dish 17 and the second cell carrier c therein can be smoothly pulled out, and the cell spheres or organoids in the concave hole array b1 are transferred to the second cell carrier c in the second culture dish 17, thereby quickly manufacturing an organoid array with controllable cell types, quantities, and densities.

[0069] The above disclosure is only a preferred embodiment of the present invention, which certainly cannot be used to limit the scope of the present invention. Therefore, equivalent changes made according to the claims of the present invention are still within the scope of the present invention.

Claims

1. A cell trap generation module for standardized and controllable regeneration of organoids, characterized in that: It includes a dehydration module, a first cell carrier, a second cell carrier, a first coated support and a second coated support; The first cell carrier is disposed in the first membrane-covered stent, the first cell carrier is provided with a concave hole array, and cells are arranged in the concave hole array; The second cell carrier is disposed in the second coated stent; A dehydration channel is provided in the dehydration module, the first film-covered support is detachably connected to the dehydration module, the first cell carrier is at least partially exposed in the dehydration channel for dehydration, the second film-covered support is arranged outside the first cell carrier, and the second cell carrier is opposite to the concave hole array of the first cell carrier, the second cell carrier enters the concave hole array and wraps the cells to form a cell trap array.

2. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 1, characterized in that: The surface of the first cell carrier opposite to the recessed hole array is exposed in the dehydration channel.

3. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 1 or 2, characterized in that: The first cell carrier and the first membrane-covered support isolate the second cell carrier from the dehydration channel.

4. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 1, characterized in that: The top surface of the second cell carrier is covered by the second membrane-covered support.

5. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 1, characterized in that: The first coated stent includes a stent body and a first supporting boss. A through hole is provided in the stent body. The first supporting boss is arranged along the circumference of the through hole. The first supporting boss is used to support the first cell carrier.

6. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 5, characterized in that: The first stent graft further comprises a support arm disposed on the periphery of the stent body; The first end of the dehydration module is provided with a mounting groove which is arranged in rotational symmetry. The mounting groove extends spirally and is arranged corresponding to the support arm. The support arm is arranged on the outer periphery of the bracket body along the spiral direction of the mounting groove. The support arm is screwed into the mounting groove, and the bracket body extends into the dehydration channel.

7. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 1, characterized in that: It also includes an array mold and a support component. The first film-covered support is detachably connected to the dehydration module. The first film-covered support and the support component cooperate to form a receiving cavity. An inert gel is provided in the receiving cavity. The inert gel is pressed by the array mold to form the concave hole array to form the first cell carrier. The first cell carrier can be pulled out of the support component along with the first film-covered support.

8. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 7, characterized in that: The support assembly includes a culture dish and a support, the support is arranged in the culture dish, the support includes a base and a second support boss formed on the base, the second support boss is adapted to the through hole, and the second support boss extends from the bottom of the through hole to form the accommodating cavity.

9. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 7, characterized in that: It also includes a dehydration platform, which is provided with a heat source cavity, an installation cavity and a dehydration hole. The installation cavity is used to accommodate the dehydration module, and the dehydration hole is used to connect the heat source cavity with the dehydration channel so that the heat source of the heat source cavity flows to the dehydration channel.

10. The cell trap generation module for standardized and controllable regeneration of organoids according to claim 9, characterized in that: The side wall of the installation cavity is provided with a first limiting groove and a second limiting groove, the second limiting groove is connected to the bottom end of the first limiting groove, and the second limiting groove is extended from the first limiting groove to the circumference of the installation cavity, and the second end of the dehydration module is provided with a mounting protrusion protruding toward the outer wall, and the mounting protrusion is inserted from the top end of the first limiting groove and screwed into the second limiting groove.

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