A cell culture apparatus, method and controller

Abstract

The application provides a cell culture device, method and controller, the cell culture device comprises a culture dish and a mixing device; the culture dish comprises a top cover, a bottom shell, and at least one intermediate shell connected between the top cover and the bottom shell in a stacked manner, a side of the intermediate shell towards the top cover is recessed to form a first accommodating groove, adjacent first accommodating grooves are communicated with each other; a side of the bottom shell close to the intermediate shell is recessed to form a second accommodating groove, and the second accommodating groove is communicated with a first accommodating groove adjacent thereto; bottoms of the first accommodating grooves and the second accommodating groove are both provided with a plurality of microwell grooves for culturing cells; the mixing device comprises a driving mechanism; the culture dish is installed on the driving mechanism, and the driving mechanism is used for driving the culture dish to translate or / and rotate, so that liquid entering the culture dish is uniformly distributed on all the microwell grooves. The application improves the uniformity of cell clusters and the efficiency of cell culture.

Description

Technical Field

[0001] This invention belongs to the field of medical device technology, and in particular relates to a cell culture device, method and controller. Background Technology

[0002] Currently, research on organoids is rapidly increasing, and organoids often require co-culturing multiple cell clusters to ensure their structural development and functional expression. Downstream applications of cell clusters include drug detection and in vivo transplantation, which generally require 10... 8 ~10 9 Reliable results can only be obtained from a few individual cells, therefore, a large number of cell clusters need to be prepared in the early stages. Given the wide range of applications of cell clusters, large-scale preparation of cell clusters is essential.

[0003] In existing technologies, a substrate is typically laid in a porous vessel, and a certain number of cells are seeded. The cells then rely on self-assembly to form clusters. However, the culture process is limited by the size of the substrate, making it difficult to obtain a large number of cell clusters, resulting in low culture efficiency. Furthermore, large-scale cell cluster culture requires multiple operations, leading to poor uniformity of the cell clusters. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a cell culture device, method and controller to address the problems of low cell cluster culture efficiency and poor uniformity in the prior art.

[0005] To address the aforementioned technical problems, embodiments of the present invention provide a cell culture apparatus, including a culture dish and a mixing device;

[0006] The petri dish includes a top cover, a bottom shell, and at least one intermediate shell stacked between the top cover and the bottom shell. The intermediate shell is recessed on one side facing the top cover to form a first receiving groove, and adjacent first receiving grooves are interconnected. The bottom shell is recessed on one side near the intermediate shell to form a second receiving groove, and the second receiving groove is interconnected with the adjacent first receiving groove.

[0007] The bottom of both the first and second accommodating grooves is provided with multiple micropores for culturing cells at intervals.

[0008] The mixing device includes a driving mechanism; the culture dish is mounted on the driving mechanism, which drives the culture dish to translate and / or rotate so that the liquid entering the culture dish is evenly distributed on all the micropore grooves.

[0009] A cell culture method, wherein the cell culture method is performed by the cell culture apparatus, the cell culture method comprising:

[0010] Liquid is injected into the first and / or second accommodating tank of the culture dish;

[0011] The drive mechanism is controlled to drive the culture dish to perform a first motion operation, so that the injected liquid is evenly distributed into all the first and second accommodating tanks;

[0012] The drive mechanism is controlled to drive the culture dish to perform a second motion operation, so that the top cover of the culture dish is located above the bottom shell, and the liquid evenly distributed in each of the first and second accommodating tanks flows into all the microporous tanks, so that the amount of liquid in all the microporous tanks is equal.

[0013] A controller includes a processor and a memory, the memory storing an executable program, the processor executing the executable program to implement the cell culture method.

[0014] In this invention, the cell culture apparatus includes a culture dish and a mixing device; the culture dish includes a top cover, a bottom shell, and at least one intermediate shell stacked between the top cover and the bottom shell, the intermediate shell having a recess on the side facing the top cover to form a first receiving groove, and adjacent first receiving grooves communicating with each other; the bottom shell having a recess on the side near the intermediate shell to form a second receiving groove, and the second receiving groove communicating with the adjacent first receiving groove; the bottom of both the first and second receiving grooves are provided with a plurality of micropores for culturing cells at intervals; the mixing device includes a driving mechanism; the culture dish is mounted on the driving mechanism, and the driving mechanism is used to drive the culture dish to translate and / or rotate, so that the liquid entering the culture dish is evenly distributed on all the micropores. This invention utilizes numerous microwells at the bottom of the first and second accommodating tanks to cultivate large cell clusters (each microwell can accommodate as many cell clusters as possible). Furthermore, by driving the culture dish to translate and / or rotate via a drive mechanism, the cell solution entering the culture dish can be evenly distributed within all the microwells of the first and second accommodating tanks, thereby improving the uniformity of the cell clusters cultured in the microwells and increasing the efficiency of cell culture. Attached Figure Description

[0015] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0016] Figure 1 This is a schematic diagram of the structure of a cell culture device provided in an embodiment of the present invention;

[0017] Figure 2 This is a schematic diagram of the structure of a petri dish provided in an embodiment of the present invention;

[0018] Figure 3 This is a schematic diagram of the structure of a petri dish provided in another embodiment of the present invention;

[0019] Figure 4 This is a schematic diagram of the structure of the intermediate shell provided in an embodiment of the present invention;

[0020] Figure 5 This is a flowchart of a cell culture method provided in an embodiment of the present invention.

[0021] The reference numerals in the accompanying drawings are as follows:

[0022] 100. Petri dish; 110. Top cover; 111. Port; 120. Bottom shell; 121. Second receiving groove; 122. Second culture plate; 123. Second outer frame; 130. Intermediate shell; 131. First receiving groove; 132. First culture plate; 133. First outer frame; 134. Through hole; 140. Micropore groove; 200. Mixing device; 210. Drive mechanism; 211. Sliding rod; 212. Telescopic rod; 213. Clamping component; 220. Base. Detailed Implementation

[0023] To make the technical problems solved, the technical solutions, and the beneficial effects of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention.

[0024] It should be understood that the terms "upper", "lower", "left", "right", "front", "rear", "middle", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing the present 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. Therefore, they should not be construed as limitations of the present invention.

[0025] like Figures 1 to 4As shown, an embodiment of the present invention provides a cell culture apparatus, including a culture dish 100 and a mixing device 200; the culture dish 100 includes a top cover 110, a bottom shell 120, and at least one intermediate shell 130 stacked between the top cover 110 and the bottom shell 120, wherein the intermediate shell 130 has a recessed first receiving groove 131 on the side facing the top cover 110, and adjacent first receiving grooves 131 are interconnected; the bottom shell 120 has a recessed second receiving groove 130 on the side near the intermediate shell 130. 21, and the second accommodating groove 121 is connected to the adjacent first accommodating groove 131; the bottom of the first accommodating groove 131 and the second accommodating groove 121 are provided with a plurality of micropore grooves 140 for culturing cells at intervals; the mixing device 200 includes a driving mechanism 210; the culture dish 100 is mounted on the driving mechanism 210, and the driving mechanism 210 is used to drive the culture dish 100 to translate and / or rotate so that the liquid entering the culture dish 100 is evenly distributed on all the micropore grooves 140.

[0026] In this embodiment, the intermediate housing 130 is connected between the top cover 110 and the bottom housing 120. One or more intermediate housings 130 can be provided as needed. When there is only one intermediate housing 130, the first receiving groove 131 of the intermediate housing 130 is directly connected to the second receiving groove 121 of the bottom housing 120. When there are multiple intermediate housings 130, adjacent intermediate housings 130 are connected sequentially, and the first receiving grooves 131 of adjacent intermediate housings 130 are interconnected. The second receiving groove 121 of the bottom housing 120 is connected to its adjacent first receiving groove 131; that is, all first receiving grooves 131 and second receiving grooves 121 are in a connected state. The microporous groove 140 opens towards the top cover 110. The shape of the microporous groove 140 can be set as needed, including but not limited to one or more of the following: spherical, rectangular, rhomboid, inverted conical, or other irregular shapes. Multiple microporous grooves 140 can have the same or different shapes.

[0027] In one embodiment, the plurality of micropores 140 are identical in shape, thereby meeting the requirement of uniformity in cell culture. The bottom of the first receiving groove 131 and / or the second receiving groove 121 is filled with the micropores 140, for example, 10,000, 15,000, and 20,000 micropores 140 are provided on the bottom of the first receiving groove 131 and / or the second receiving groove 121. The culture dish 100 includes a plurality of receiving grooves (a second receiving groove 121 and at least one first receiving groove 131), therefore, when the culture dish 100 is normally placed (i.e., as...), Figure 3When the top cover 110 is located at the top and the bottom shell 120 is located at the bottom, under the influence of gravity and the position of the connecting channel between the second accommodating groove 121 and the first accommodating groove 131, the liquid in the second accommodating groove 121 and the first accommodating groove 131 may not be evenly distributed in the second accommodating groove 121 and the first accommodating groove 131. Therefore, the driving mechanism 210 is used to drive the culture dish 100 to translate and / or rotate, so that the liquid entering the culture dish 100 can be evenly distributed in the second accommodating groove 121 and the first accommodating groove 131, and then evenly distributed on all the microporous grooves 140.

[0028] This invention utilizes numerous micropores 140 at the bottom of the first and second accommodating tanks 131 and 121 to cultivate large cell clusters (each micropore can accommodate as many cell clusters as possible). Furthermore, by driving the culture dish 100 to translate and / or rotate via the drive mechanism 210, the cell solution entering the culture dish 100 is evenly distributed within all the micropores 140 of the first and second accommodating tanks 131 and 121, thereby improving the uniformity of the cell clusters cultured in the micropores 140 and increasing the efficiency of cell culture.

[0029] In one embodiment, the first receiving tank 131 and the second receiving tank 121 have the same capacity. In this embodiment, the petri dish 100 is driven to rotate by the driving mechanism 210 until the first receiving tank 131 and the second receiving tank 121 are in a vertical state. At this time, the positions where the first receiving tank 131 and the second receiving tank 121 are connected are located at the bottom positions corresponding to the vertical first receiving tank 131 and the second receiving tank 121. Therefore, the liquid will flow through the connected positions until the liquid in the first receiving tank 131 and the second receiving tank 121 is evenly distributed.

[0030] In one embodiment, the top cover 110 and the bottom shell 120 of the culture dish 100 are respectively provided with mounting portions for mounting on the drive mechanism 210. This makes the culture dish 100 more stable when mounted on the drive mechanism 210.

[0031] like Figure 1As shown, in one embodiment, the mixing device 200 includes a base 220; the driving mechanism 210 includes a sliding rod 211, a telescopic rod 212, and a clamping member 213 for mounting the culture dish 100; the clamping member 213 is rotatably connected to the first end of the telescopic rod 212, and the clamping member 213 is used to drive the culture dish 100 to rotate around a first reference line; the first reference line is parallel to the telescopic rod 212; the second end of the telescopic rod 212 is rotatably mounted on the sliding rod 211, and the telescopic rod 212 is used to drive the culture dish 100 to rotate around a first reference line; the first reference line is parallel to the telescopic rod 212; the second end of the telescopic rod 212 is rotatably mounted on the sliding rod 211, and the telescopic rod 212 is used to drive the culture dish 100 to rotate around a first reference line; the first reference line is parallel to the telescopic rod 212; the second end of the telescopic rod 212 is rotatably mounted on the sliding rod 211, and the telescopic rod 212 is used to drive the culture dish 100 to rotate around a first reference line; the first reference line is parallel to the telescopic rod 212; the second end of the telescopic rod 212 is rotatably mounted on the sliding rod 211, and the second end of ... second end of the telescopic rod 212 is rotatably mounted on the sliding rod 211 The clamping member 213 and the culture dish 100 are moved along the first reference line and / or rotated around the second reference line; the second reference line is perpendicular to the first reference line and parallel to the horizontal plane (the second reference line can be set parallel to the sliding rod 211); the sliding rod 211 is slidably mounted on the base 220, and the sliding rod 211 is used to move the telescopic rod, the clamping member 213, and the culture dish 100 along the third reference line, which is perpendicular to both the first and second reference lines. It is understood that the clamping member 213 is a height-adjustable structure (e.g., including an upper gripping plate, a lower gripping plate, and a telescopic rod that can be telescopically connected between the upper and lower gripping plates), and the clamping width can be adjusted according to the height of the culture dish 100. That is, when there are one or more intermediate shells 130, the clamping member 213 can clamp the culture dish 100. Each of the sliding rod 211, the telescopic rod 212, and the clamping member 213 is equipped with a driving component (the driving components corresponding to the sliding rod 211, the telescopic rod 212, and the clamping member 213 can be different motors, drive cylinders, etc.). The driving component is used to drive the clamping member 213 to clamp and release, the telescopic rod 212 to extend and retract along the first reference line, the telescopic rod 212 to rotate around the second reference line, and the sliding rod 211 to translate along the third reference line.

[0032] like Figures 2 to 4As shown, the intermediate shell 130 includes a first culture plate 132 and a first outer frame 133 surrounding the first culture plate 132; the first receiving groove 131 is formed by the first outer frame 133 and the first culture plate 132; the bottom shell 120 includes a second culture plate 122 and a second outer frame 123 surrounding the second culture plate 122; the second receiving groove 121 is formed by the second outer frame 123 and the second culture plate 122; the microporous groove 140 is disposed on the first culture plate 132 and the second culture plate 122; the first culture plate 132 and / or the first outer frame 133 are provided with through holes 134; adjacent first receiving grooves 131 and the first receiving groove 131 and the second receiving groove 121 are interconnected through the through holes 134. Understandably, the first culture plate 132 can be disposed at the bottom of the first receiving groove 131 by means of connection (e.g., bonding, snap-fitting, or integral molding), in which case the microporous groove 140 is disposed on the first culture plate 132 (i.e., at the bottom of the first receiving groove 131). The second culture plate 122 can be disposed at the bottom of the second receiving groove 121 by means of connection (e.g., bonding, snap-fitting, or integral molding), in which case the microporous groove 140 is disposed on the second culture plate 122 (i.e., at the bottom of the second receiving groove 121). The through hole 134 can be disposed on the first culture plate 132, or on the first outer frame 133, or simultaneously on both the first culture plate 132 and the first outer frame 133, as long as the first receiving groove 131 and the second receiving groove 121 are connected through the through hole 134.

[0033] In one embodiment, the through hole 134 is disposed on the first culture plate 132 near one end of the first outer frame 133 (i.e., at the edge of the first receiving groove 131). When the driving mechanism 210 drives the culture dish 100 to rotate to a vertical position, the through hole 134 can be rotated to the bottom position of the culture dish 100. This allows the liquid to submerge the through hole 134 at the bottom position of the culture dish 100 when the liquid in the second receiving groove 121 and the first receiving groove 131 is low, i.e., when the liquid level is low. Thus, the liquid in the second receiving groove 121 and the first receiving groove 131 is connected through the through hole 134, thereby making the liquid evenly distributed in the second receiving groove 121 and the first receiving groove 131.

[0034] In another embodiment, the through hole 134 can also be provided on the first outer frame 133 (i.e., at the edge of the first receiving groove 131). Similarly, when the driving mechanism 210 drives the culture dish 100 to rotate to a vertical state, so that the through hole 134 is rotated to the bottom position of the culture dish 100, even if the liquid in the second receiving groove 121 and the first receiving groove 131 is small, the liquid can submerge the through hole 134, thereby connecting the liquid in the second receiving groove 121 and the first receiving groove 131 through the through hole 134, and thus making the liquid evenly distributed in the second receiving groove 121 and the first receiving groove 131.

[0035] In one embodiment, the top cover 110 is provided with a port 111 communicating with the first receiving groove 131; the port 111 is located in the middle of the top cover 110. Understandably, cell fluid can be injected into each of the first receiving grooves 131 and / or the second receiving groove 121 through the port 111 in the middle of the top cover 110. Then, through translation and / or rotation, cell fluid can be evenly distributed in all the micropores 140 on each of the first receiving grooves 131 and / or the second receiving groove 121. This allows the cell culture device to prepare large quantities of uniform cell clusters and also improves the convenience of adding cell fluid to the culture dish 100. In one embodiment, the centerline of the port 111 is inclined towards the through hole 134. Understandably, the centerline of the port 111 is inclined toward the through hole 134, which allows liquid (e.g., cell fluid) injected from the port 111 into the first receiving tank 131 to pass through the through hole 134 more quickly into the second receiving tank 121, thereby reducing the residue of cell fluid.

[0036] like Figure 2 and Figure 3 As shown, in one embodiment, the top cover 110 is provided with a port 111 communicating with the first receiving groove 131; the top cover 110 is rectangular, and the port 111 includes a first opening and a second opening, the first opening and the second opening being located at two adjacent corners of the rectangular top cover 110. It is understood that, preferably, the inner diameter of the first opening is larger than the inner diameter of the second opening. Thus, a large flow rate of cell solution can be injected into the culture dish 100 through the larger inner diameter first opening, and a small flow rate of cell solution can be injected into the culture dish 100 through the smaller inner diameter second opening; and the first and second openings communicate with the outside, so that after the cell solution is injected, oxygen, etc., can also be injected into the culture dish 100 through the first opening and / or the second opening.

[0037] In one embodiment, the through-hole 134 includes a first flow hole and a second flow hole. The first flow hole is located below the first opening, and the second flow hole is located below the second opening. Understandably, the first opening and the second opening are located at two adjacent corners of the rectangular top cover 110, and therefore, the first flow hole and the second flow hole are also located at two adjacent corners of the first culture plate 132. Thus, when the driving mechanism 210 drives the culture dish 100 to a vertical position, the first flow hole and the second flow hole are simultaneously rotated to the bottom position of the culture dish 100. The liquid in the first receiving groove 131 and the second receiving groove 121 can simultaneously communicate through the first flow hole and the second flow hole, thereby accelerating the liquid flow rate and improving efficiency. In one embodiment, the centerline of the port 111 is inclined towards the through-hole 134. Understandably, in this embodiment, the inclination of the centerline of the port 111 toward the through hole 134 means that the centerline of the first opening is inclination toward the first flow hole, and the centerline of the second opening is inclination toward the second flow hole. In this way, when liquid (e.g., cell fluid) is injected into the first receiving tank 131 from the first opening and the second opening through the first flow hole and the second flow hole respectively, it can enter the second receiving tank 121 more quickly through the first flow hole and the second flow hole, thereby reducing the residue of cell fluid.

[0038] In one embodiment, the petri dish 100 further includes a filter membrane installed in the port 111. Understandably, the filter membrane includes, but is not limited to, a sterile filter membrane, an air filter membrane, etc. The filter membrane prevents bacteria and impurities from the external environment from entering the petri dish 100. When the port 111 is located in the middle of the top cover 110, it prevents the filter membrane in the port 111 from being immersed in the liquid in the petri dish 100 when the petri dish 100 is rotated, thus avoiding affecting the filtration effect of the filter membrane.

[0039] In one embodiment, the surface of the microporous groove 140 is covered with a hydrophobic coating (not shown). That is, the hydrophobic coating covers the groove wall of the microporous groove 140, so that the hydrophobic properties of the hydrophobic coating facilitate the suspension culture of cells in the cell fluid entering the microporous groove 140.

[0040] In one embodiment, the cross-sectional area of ​​the microporous groove 140 gradually decreases from the top cover 110 toward the bottom shell 120. Understandably, the gradual decrease in the cross-sectional area of ​​the microporous groove 140, i.e., the gradual downward tilt and condensation of the microporous groove 140, allows cells in the cell sap (with a cell density greater than that in the cell sap) to descend to the bottom of the microporous groove 140 without additional centrifugal force.

[0041] In one embodiment, the micropore groove 140 is a conical groove. Understandably, the micropore groove 140 can be a square pyramidal groove or a hexagonal pyramidal groove, etc. In this embodiment, the top of the micropore groove 140 has a sharp edge, which can prevent cells from settling and differentiating at the edge of the pore opening of the micropore groove 140, and the micropore groove 140 has a simple structure and low manufacturing cost. In another specific embodiment, the micropore groove 140 is a conical groove. Understandably, the inner wall of the micropore groove 140 is a smooth curved surface, which can further promote cell aggregation and growth.

[0042] In one embodiment, the depth of the microporous groove 140 is 150 μm to 550 μm. It is understood that the depth of the microporous groove 140 can be set to 150 μm, 250 μm, 350 μm, 450 μm, or 550 μm, etc., according to actual needs.

[0043] In one embodiment, the groove opening area of ​​the microporous groove 140 is [area missing]. Understandably, the cross-sectional area of ​​the top of the microporous groove 140 can be set to 0.1 [area missing] according to actual needs. 0.2 0.3 0.4 Or 0.5 wait.

[0044] like Figure 5 As shown, an embodiment of the present invention also provides a cell culture method, wherein the cell culture method is performed by the aforementioned cell culture apparatus, and the cell culture method includes:

[0045] S10. Inject liquid into the first receiving tank 131 and / or the second receiving tank 121 of the culture dish 100. Understandably, the culture dish 100 is provided with an opening for injecting liquid (e.g., cell sap or nutrient solution), through which liquid can be injected into the first receiving tank 131 and / or the second receiving tank 121. The opening can be located on the top cover 110, the intermediate shell 130, or the bottom shell 120. In one embodiment, the opening refers to a port 111 on the top cover 110 that communicates with the first receiving tank 131, through which liquid (e.g., cell sap or nutrient solution) can be injected into the first receiving tank 131 and / or the second receiving tank 121.

[0046] S20. Control the drive mechanism 210 to drive the culture dish 100 to perform a first motion operation, so that the injected liquid is evenly distributed into all the first receiving tanks 131 and the second receiving tanks 121. The first motion operation includes, but is not limited to, rotation, and may also include, for example, translation in the up, down, left, and right directions.

[0047] In one embodiment, the top cover 110 is provided with a port 111 communicating with the first receiving groove 131; step S20 includes:

[0048] The drive mechanism 210 is controlled to rotate the culture dish 100 in a first direction (i.e., the rotation direction in which the through hole 134 can be located at the bottom), so that the through hole 134 is located at the bottom of the culture dish 100. After the drive mechanism 210 rotates the culture dish 100 so that the through hole 134 is located at the bottom of the culture dish 100, the first receiving groove 131 and the second receiving groove 121 are connected through the through hole 134. At this time, the liquid in the first receiving groove 131 and the second receiving groove 121 will flow to the same horizontal plane through the through hole 134. In one embodiment, the first receiving groove 131 and the second receiving groove 121 have the same capacity; in this case, controlling the drive mechanism 210 to rotate the culture dish 100 will make the first receiving groove 131 and the second receiving groove 121 of the culture dish 100 in a vertical state. At this time, the liquid in the first accommodating tank 131 and the second accommodating tank 121 will flow to the same horizontal plane through the through hole 134, and the liquid will be evenly distributed in the first accommodating tank 131 and the second accommodating tank 121.

[0049] In one embodiment, the top cover 110 is provided with a port 111 communicating with the first receiving groove 131; and the through hole 134 includes a first flow hole and a second flow hole, the first flow hole and the second flow hole being located at two adjacent corners of the first culture plate 132 respectively; step S20 includes:

[0050] The drive mechanism 210 is controlled to rotate the culture dish 100 in a second direction (i.e., a rotational direction in which the first flow hole and the second flow hole can be located at the bottom), so that both the first flow hole and the second flow hole are located at the bottom of the culture dish 100. Understandably, the first flow hole and the second flow hole are located at two adjacent corners of the first culture plate 132, so that when the drive mechanism 210 drives the culture dish 100 to rotate to a vertical position, the first flow hole and the second flow hole simultaneously rotate to the bottom of the culture dish 100. Furthermore, the liquid in the first receiving tank 131 and the second receiving tank 121 can simultaneously communicate through the first flow hole and the second flow hole, which can accelerate the liquid flow rate and improve efficiency.

[0051] The liquid level sensor can be used to detect whether the liquid in the first container 131 and the second container 121 are at the same level.

[0052] S30. Control the drive mechanism 210 to drive the culture dish 100 to perform a second motion operation, so that the top cover 110 of the culture dish 100 is positioned above the bottom shell 120, and the liquid evenly distributed in each of the first receiving grooves 131 and the second receiving grooves 121 flows into all the microporous grooves 140, making the amount of liquid in all the microporous grooves 140 equal. The second motion operation includes, but is not limited to, rotation. Specifically, step S30 includes:

[0053] Step S30 includes:

[0054] S31, after the liquid in the first receiving tank 131 and the second receiving tank 121 of the culture dish 100 flows to the same horizontal plane through the first flow hole and the second flow hole, the driving mechanism 210 is controlled to drive the culture dish 100 to rotate in a third direction (understandably, the third direction should be opposite to the rotation direction in step S20. For example, if step S20 controls the driving mechanism 210 to drive the culture dish 100 to rotate in a first direction, then the third direction should be opposite to the first direction; and if step S20 controls the driving mechanism 210 to drive the culture dish 100 to rotate in a second direction, then the third direction should be opposite to the second direction) so that the top cover 110 of the culture dish 100 is located above the bottom shell 120, and the first culture plate 132 and the second culture plate 122 are both in a horizontal position. Understandably, once the first culture plate 132 and the second culture plate 122 are both in a horizontal position, the liquid in the first accommodating tank 131 and the second accommodating tank 121 will flow into all the microporous tanks 140 under the action of gravity.

[0055] In step S20, after the liquid in the first receiving tank 131 and the second receiving tank 121 of the culture dish 100 flows to the same horizontal plane through the through hole 134, in step S30, the driving mechanism 210 is further controlled to rotate the culture dish 100 so that the first culture plate 132 and the second culture plate 122 are both in a horizontal position (and at this time, the top cover 110 of the culture dish 100 must be above the bottom shell 120). It can be understood that after the first culture plate 132 and the second culture plate 122 are both in a horizontal position, the liquid evenly distributed in each of the first receiving tanks 131 and the second receiving tanks 121 will flow into all the microporous tanks 140 under the action of gravity, making the amount of liquid in all the microporous tanks 140 equal. The liquid level sensor can be used to detect whether the liquid in the microporous tanks 140 is at the same horizontal plane.

[0056] S32, control the drive mechanism 210 to drive the culture dish 100 to reciprocate and translate, so that the liquid adhering to the bottom of the top cover 110 or the first receiving tank 131 drops down faster (drops into the first receiving tank 131 located below the top cover 110, or another first receiving tank 131 or the second receiving tank 121 located below the first receiving tank 131). That is, since the liquid (such as cell fluid) is viscous, when the top cover 110 of the culture dish 100 is above the bottom shell 120, a small amount of liquid may still adhere to the bottom of the top cover 110 or the first receiving tank 131. At this time, controlling the drive mechanism 210 to drive the culture dish 100 to swing back and forth in the horizontal direction can prevent the liquid in the first receiving tank 131 from flowing from the through hole 134 into the second receiving tank 121, while making the liquid at the bottom of the first receiving tank 131 drip into the lower first receiving tank 131 or the second receiving tank 121 faster in the reciprocating swing motion, thereby improving efficiency.

[0057] It is understood that the cell culture method described above is not limited to that described in the above embodiments. The specific settings of the cell culture method of the present invention correspond one-to-one with the cell culture device described above, and will not be repeated here.

[0058] An embodiment of the present invention also provides a controller, the controller including a processor and a memory, the memory storing an executable program, and the processor being used to execute the executable program to implement the cell culture method.

[0059] The specific configuration of the controller of this invention corresponds one-to-one with the above-described cell culture method, and will not be repeated here. The controller is connected to the drive mechanism 210 of the cell culture apparatus. Furthermore, the controller can control the drive mechanism 210 to drive the culture dish 100 to translate and / or rotate, so that the liquid entering the culture dish 100 is evenly distributed on all the micropores 140. Each module in the controller can be implemented entirely or partially through software, hardware, or a combination thereof. Each module can be embedded in the controller in hardware form or independent of the controller, or it can be stored in the controller in software form, so that the controller can call and execute the operations corresponding to each module.

[0060] An embodiment of the present invention also provides a computer-readable storage medium storing a computer program that, when executed by a processor, implements the above-described cell culture method.

[0061] Those skilled in the art will understand that all or part of the processes in the above embodiments can be implemented by instructing related hardware with computer-readable instructions. These computer-readable instructions can be stored in a non-volatile readable storage medium or a volatile readable storage medium. When executed, these computer-readable instructions can include the processes of the embodiments of the above methods. Any references to memory, storage, databases, or other media used in the embodiments provided by this invention can include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM), or flash memory. Volatile memory may include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in a variety of forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), dual data rate SDRAM (DDRSDRAM), enhanced SDRAM (ESDRAM), synchronous link DRAM (SLDRAM), RAMbus direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and memory bus dynamic RAM (RDRAM), etc.

[0062] Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the above-described division of functional units and modules is used as an example. In practical applications, the above functions can be assigned to different functional units and modules as needed, that is, the internal structure of the system can be divided into different functional units or modules to complete all or part of the functions described above.

[0063] The above are merely embodiments of the cell culture apparatus, method, controller, and medium of the present invention, and are not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A cell culture device, characterized in that, Includes petri dishes and mixing devices; The petri dish includes a top cover, a bottom shell, and at least one intermediate shell stacked between the top cover and the bottom shell. The intermediate shell is recessed on one side facing the top cover to form a first receiving groove, and adjacent first receiving grooves are interconnected. The bottom shell is recessed on one side near the intermediate shell to form a second receiving groove, and the second receiving groove is interconnected with the adjacent first receiving groove. The bottom of both the first and second accommodating grooves is provided with multiple micropores for culturing cells at intervals. The mixing device includes a driving mechanism; the culture dish is mounted on the driving mechanism, and the driving mechanism is used to drive the culture dish to translate and / or rotate so that the liquid entering the culture dish is evenly distributed on all the micropore grooves; The mixing device includes a base; the driving mechanism includes a sliding rod, a telescopic rod, and a clamping component for mounting the culture dish; The clamping member is rotatably connected to the first end of the telescopic rod, and the clamping member is used to drive the culture dish to rotate around a first reference line; the first reference line is parallel to the telescopic rod; The second end of the telescopic rod is rotatably mounted on the sliding rod, and the telescopic rod is used to drive the clamping member and the culture dish to translate along the first reference line and / or rotate around the second reference line; the second reference line is perpendicular to the first reference line and parallel to the horizontal plane; The sliding rod is slidably mounted on the base, and the sliding rod is used to drive the telescopic rod, the clamping member and the culture dish to translate along the third reference line, which is perpendicular to both the first reference line and the second reference line.

2. The cell culture apparatus according to claim 1, characterized in that, The intermediate shell includes a first culture plate and a first outer frame surrounding the first culture plate; the first accommodating groove is formed by the first outer frame and the first culture plate; the bottom shell includes a second culture plate and a second outer frame surrounding the second culture plate; the second accommodating groove is formed by the second outer frame and the second culture plate; the microporous groove is disposed on the first culture plate and the second culture plate; The first culture plate and / or the first outer frame are provided with through holes; adjacent first receiving slots and the first receiving slot and the second receiving slot are interconnected through the through holes.

3. The cell culture apparatus according to claim 2, characterized in that, The top cover has a port that communicates with the first receiving slot; the port is located in the middle of the top cover.

4. The cell culture apparatus according to claim 2, characterized in that, The top cover is provided with a port communicating with the first receiving groove; the top cover is rectangular, and the port includes a first opening and a second opening, the first opening and the second opening being located at two adjacent corners of the rectangular top cover respectively.

5. The cell culture apparatus according to claim 3 or 4, characterized in that, The centerline of the port is inclined toward the through hole.

6. The cell culture apparatus according to claim 3 or 4, characterized in that, The petri dish also includes a filter membrane installed in the port.

7. The cell culture apparatus according to claim 1, characterized in that, The surface of the microporous groove is covered with a hydrophobic coating.

8. A cell culture method, characterized in that, The cell culture method is performed using the cell culture apparatus as described in any one of claims 1 to 7, and the cell culture method includes: Liquid is injected into the first and / or second accommodating tank of the culture dish; The drive mechanism is controlled to drive the culture dish to perform a first motion operation, so that the injected liquid is evenly distributed into all the first and second accommodating tanks; The drive mechanism is controlled to drive the culture dish to perform a second motion operation, so that the top cover of the culture dish is located above the bottom shell, and the liquid evenly distributed in each of the first and second accommodating tanks flows into all the microporous tanks, so that the amount of liquid in all the microporous tanks is equal.

9. A controller, characterized in that, The controller includes a processor and a memory, the memory storing an executable program, and the processor executing the executable program to implement the cell culture method as described in claim 8.

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