Cell culture device, method and controller

Abstract

The invention provides a cell culture device, a cell culture method and a 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 middle shell connected between the top cover and the bottom shell in a stacked mode, the side, facing the top cover, of each middle shell is sunken to form a first containing groove, and every two adjacent first containing grooves communicate with each other; the side, close to the middle shell, of the bottom shell is recessed to form a second containing groove, and the second containing groove is communicated with the first containing groove adjacent to the second containing groove. A plurality of micropore grooves for culturing cells are formed in the bottoms of the first accommodating groove and the second accommodating groove at intervals; the uniform mixing device comprises a driving mechanism; the culture dish is mounted 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 micropore grooves. According to the invention, the uniformity of cell clusters and the efficiency of cell culture are improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical devices, and in particular relates to a cell culture device, method and controller. Background Art

[0002] At present, research on organoids is increasing rapidly, and organoids often require co-culture of multiple cells to ensure their structural development and functional expression. The downstream applications of cell clusters include drug testing, in vivo transplantation, etc. These downstream applications generally require 10 8 ~10 9 In order to obtain reliable results, a large number of cell clusters need to be prepared in the early stage. Given the wide application of cell clusters, large-scale preparation of cell clusters is very necessary.

[0003] In the prior art, a substrate is generally laid in a vessel with holes, and a certain number of cells are planted and then rely on cell self-assembly to form clusters. In this way, it is difficult to obtain a large number of cell clusters during the culture process of cell clusters due to the limitation of the substrate size, and the culture efficiency is low. In addition, the culture of large-scale cell clusters requires multiple operations, so the uniformity of the cell clusters is poor. Summary of the invention

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

[0005] In order to solve the above technical problems, an embodiment of the present invention provides a cell culture device, including a culture dish and a mixing device;

[0006] The culture dish comprises a top cover, a bottom shell, and at least one intermediate shell stacked and connected between the top cover and the bottom shell, wherein the intermediate shell is recessed on one side facing the top cover to form a first receiving groove, and adjacent first receiving grooves are connected to each other; the bottom shell is recessed on one side close to the intermediate shell to form a second receiving groove, and the second receiving groove is connected to the first receiving groove adjacent thereto;

[0007] The bottom of the first containing tank and the bottom of the second containing tank are both provided with a plurality of microporous grooves for culturing cells at intervals;

[0008] The mixing device comprises 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 microporous grooves.

[0009] A cell culture method, the cell culture method is performed by the cell culture device, and the cell culture method comprises:

[0010] Injecting liquid into the first containing tank and / or the second containing tank of the culture dish;

[0011] Controlling the driving mechanism to drive the culture dish to perform a first movement operation so that the injected liquid is evenly distributed to all the first containing grooves and the second containing grooves;

[0012] The driving mechanism is controlled to drive the culture dish to perform a second movement 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 containing grooves and the second containing grooves flows into all the microporous grooves, and the amount of liquid in all the microporous grooves is equal.

[0013] A controller comprises a processor and a memory, wherein the memory stores an executable program, and the processor is used to execute the executable program to implement the cell centrifugation method.

[0014] In the present invention, the cell culture device 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 and connected 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 connected to each other; the bottom of the bottom shell close to the intermediate shell is recessed to form a second receiving groove, and the second receiving groove is connected to the first receiving groove adjacent to it; the bottoms of the first receiving groove and the second receiving groove are both spaced apart with a plurality of microporous grooves for culturing cells; 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 microporous grooves. The present invention can culture a large number of cell clusters (cell clusters can be cultured in each microporous groove) by setting a large number of microporous grooves at the bottom of the first containing groove and the second containing groove; and by driving the culture dish to translate and / or rotate through the driving mechanism, the cell fluid entering the culture dish can be evenly distributed in all the microporous grooves on the first containing groove and the second containing groove, thereby improving the uniformity of the cell clusters cultured in the microporous grooves and also improving the efficiency of cell culture. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0016] Figure 1 is a schematic structural diagram of a cell culture device provided by one embodiment of the present invention;

[0017] Figure 2 is a schematic structural diagram of a culture dish provided by an embodiment of the present invention;

[0018] Figure 3 is a schematic structural diagram of a culture dish provided by another embodiment of the present invention;

[0019] Figure 4 is a schematic structural diagram of an intermediate housing provided by an embodiment of the present invention;

[0020] Figure 5 It is a flow chart of a cell culture method provided by one embodiment of the present invention.

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

[0022] 100, culture dish; 110, top cover; 111, port; 120, bottom shell; 121, second accommodating groove; 122, second culture plate; 123, second outer frame; 130, intermediate shell; 131, first accommodating groove; 132, first culture plate; 133, first outer frame; 134, through hole; 140, microporous groove; 200, mixing device; 210, driving mechanism; 211, sliding rod; 212, telescopic rod; 213, clamping member; 220, base. DETAILED DESCRIPTION

[0023] In order to make the technical problems, technical solutions and beneficial effects solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0024] It should be understood that the directions or positional relationships indicated by terms such as "upper", "lower", "left", "right", "front", "back", and "middle" are based on the directions or positional relationships shown in the accompanying drawings and are only for the convenience of describing the present invention and simplifying the description. They do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore should not be understood as a limitation of the present invention.

[0025] like Figures 1 to 4As shown, a cell culture device provided by an embodiment of the present invention includes 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 and connected between the top cover 110 and the bottom shell 120, the intermediate shell 130 is recessed on one side toward the top cover 110 to form a first receiving groove 131, and adjacent first receiving grooves 131 are connected to each other; the bottom shell 120 is recessed on one side close to the intermediate shell 130 to form a second receiving groove 131. 21, and the second containing groove 121 is connected with the first containing groove 131 adjacent thereto; the bottoms of the first containing groove 131 and the second containing groove 121 are both spaced apart with a plurality of microporous grooves 140 for culturing cells; 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 microporous grooves 140.

[0026] In this embodiment, the intermediate shell 130 is connected between the top cover 110 and the bottom shell 120. One or more intermediate shells 130 can be provided as needed. When the number of the intermediate shell 130 is one, the first receiving groove 131 of the intermediate shell 130 is directly connected to the second receiving groove 121 of the bottom shell 120; when the number of the intermediate shell 130 is multiple, the adjacent intermediate shells 130 are connected in sequence, and the first receiving grooves 131 of the adjacent intermediate shells 130 are connected to each other, and the second receiving groove 121 of the bottom shell 120 is connected to the first receiving groove 131 adjacent to it, that is, all the first receiving grooves 131 and the second receiving grooves 121 are connected. The microporous groove 140 opens toward the top cover 110, and the shape of the microporous groove 140 can be set as needed, including but not limited to one or more of spherical, rectangular, rhombus, inverted cone, or other irregular shapes; the shapes of multiple microporous grooves 140 can be the same or different.

[0027] In one embodiment, the shapes of the plurality of microporous grooves 140 are the same, so as to meet the uniformity requirement in cell culture. The bottom of the first receiving groove 131 and / or the second receiving groove 121 is covered with the microporous grooves 140, for example, 10,000, 15,000, and 20,000 microporous grooves 140 are arranged 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 (the second receiving groove 121 and at least one first receiving groove 131), so when the culture dish 100 is placed normally (i.e., as shown in FIG. 1 ), the microporous grooves 140 are arranged on the bottom of the first receiving groove 131 and / or the second receiving groove 121. Figure 3As shown, when 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 receiving groove 121 and the first receiving groove 131, the liquid in the second receiving groove 121 and the first receiving groove 131 may not be evenly distributed in the second receiving groove 121 and the first receiving 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 receiving groove 121 and the first receiving groove 131, and then evenly distributed on all the microporous grooves 140.

[0028] The present invention can culture a large number of cell clusters (cell clusters can be cultured in each microporous groove) by setting a large number of microporous grooves 140 at the bottom of the first containing groove 131 and the second containing groove 121; and by driving the culture dish 100 to translate and / or rotate through the driving mechanism 210, the cell fluid entering the culture dish 100 can be evenly distributed in all the microporous grooves 140 on the first containing groove 131 and the second containing groove 121, thereby improving the uniformity of the cell clusters cultured in the microporous grooves 140 and also improving the efficiency of cell culture.

[0029] In one embodiment, the first receiving groove 131 and the second receiving groove 121 have the same capacity. In this embodiment, the culture dish 100 is driven to rotate by the driving mechanism 210 until the first receiving groove 131 and the second receiving groove 121 are in a vertical state. At this time, the position where the first receiving groove 131 and the second receiving groove 121 are connected to each other is located at a position corresponding to the bottom of the vertical first receiving groove 131 and the second receiving groove 121. Therefore, the liquid will flow through the interconnected position until the liquid in the first receiving groove 131 and the second receiving groove 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 being mounted on the driving mechanism 210 , so that the culture dish 100 is more stable when mounted on the driving 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 The clamping member 213 and the culture dish 100 are used to translate along the first reference line or / and rotate 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 arranged in parallel with the sliding rod 211); the sliding rod 211 is slidably mounted on the base 220, and the sliding rod 211 is used to drive the telescopic member, the clamping member 213 and the culture dish 100 to translate along the third reference line, and the third reference line is perpendicular to the first reference line and the second reference line. It can be understood that the clamping member 213 is a height-adjustable structure (for example, including an upper gripping plate, a lower gripping plate, and a telescopic member that can be telescopically connected between the upper gripping plate and the lower gripping plate), and the clamping width can be adjusted according to the height of the culture dish 100. That is, when the intermediate housing 130 is one or more, the clamping member 213 can clamp the culture dish 100. The sliding rod 211, the telescopic rod 212 and the clamping member 213 are all provided with a driving member (the driving members corresponding to the sliding rod 211, the telescopic rod 212 and the clamping member 213 can be different motors, drive cylinders, etc.). The driving member 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 arranged around the first culture plate 132; the first receiving groove 131 is surrounded 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 arranged around the second culture plate 122; the second receiving groove 121 is surrounded by the second outer frame 123 and the second culture plate 122; the microporous groove 140 is arranged 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 a through hole 134; adjacent first receiving grooves 131, and the first receiving groove 131 and the second receiving groove 121 are interconnected through the through hole 134. It can be understood that the first culture plate 132 can be set at the bottom of the first containing groove 131 by a connection method (such as bonding, snap-fitting or integral molding, etc.), and in this case, the microporous groove 140 is set on the first culture plate 132 (that is, the bottom of the first containing groove 131). The second culture plate 122 can be set at the bottom of the second containing groove 121 by a connection method (such as bonding, snap-fitting or integral molding, etc.), and in this case, the microporous groove 140 is set on the second culture plate 122 (that is, the bottom of the second containing groove 121). The through hole 134 can be set on the first culture plate 132, the through hole 134 can also be set on the first outer frame 133, and the through hole 134 can also be set on the first culture plate 132 and the first outer frame 133 at the same time, as long as the first containing grooves 131 and the first containing grooves 131 and the second containing grooves 121 can be connected through the through hole 134.

[0033] In one embodiment, the through hole 134 is arranged on the first culture plate 132 at one end close to the first outer frame 133 (that is, the edge position of the first receiving groove 131), so that when the driving mechanism 210 drives the culture dish 100 to rotate to a vertical state, the through hole 134 can be rotated to the bottom position of the culture dish 100, so that when the liquid in the second receiving groove 121 and the first receiving groove 131 is less, that is, when the liquid level is low, the liquid can also submerge the through hole 134 located at the bottom position of the culture dish 100, so that the liquid in the second receiving groove 121 and the first receiving groove 131 are connected through the through hole 134, and the liquid is 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 set on the first outer frame 133 (that is, the edge position of the first receiving groove 131), and the driving mechanism 210 can also drive the culture dish 100 to rotate to a vertical state, so that when the through hole 134 is rotated to the bottom position of the culture dish 100, even if there is less liquid in the second receiving groove 121 and the first receiving groove 131, the liquid can submerge the through hole 134, and then the liquid in the second receiving groove 121 and the first receiving groove 131 are connected through the through hole 134, so that the liquid is 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 connected to the first receiving groove 131; the port 111 is located in the middle of the top cover 110. It can be understood that the cell solution can be injected into each of the first receiving grooves 131 and / or the second receiving grooves 121 through the port 111 in the middle of the top cover 110, and then through operations such as translation and / or rotation, the cell solution can be evenly distributed in all the microporous grooves 140 on each of the first receiving grooves 131 and / or the second receiving grooves 121, so that the cell culture device can prepare a large number of uniform cell clusters, and the convenience of adding cell solution to the culture dish 100 is also improved. In one embodiment, the center line of the port 111 is inclined toward the through hole 134. It can be understood that the center line of the port 111 is inclined toward the through hole 134, so that when the liquid (such as cell fluid) is injected into the first receiving groove 131 from the port 111, it can enter the second receiving groove 121 through the through hole 134 more quickly, thereby reducing the residual cell fluid.

[0036] like Figure 2 and Figure 3 As shown, in one embodiment, the top cover 110 is provided with a port 111 connected to the first receiving groove 131; the top cover 110 is rectangular, and the port 111 includes a first opening and a second opening, and the first opening and the second opening are respectively located at two adjacent corners of the rectangular top cover 110. It can be understood that, as a preference, the inner diameter of the first opening is greater than the inner diameter of the second opening. Thus, a large flow of cell fluid can be injected into the culture dish 100 through the first opening with a larger inner diameter, and a small flow of cell fluid can be injected into the culture dish 100 through the second opening with a smaller inner diameter; and the first opening and the second opening are connected to the outside, so that after the cell fluid is injected, oxygen and the like can 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, wherein the first flow hole is arranged below the first opening, and the second flow hole is arranged below the second opening. It can be understood that the first opening and the second opening are respectively located at two adjacent corners of the rectangular top cover 110, and therefore, the first flow hole and the second flow hole are also respectively located at two adjacent corners of the first culture plate 132. Thus, when the driving mechanism 210 drives the culture dish 100 to rotate to a vertical state, the first flow hole and the second flow hole can be rotated to the bottom position of the culture dish 100 at the same time, and the liquid in the first receiving groove 131 and the second receiving groove 121 can be connected through the first flow hole and the second flow hole at the same time, thereby accelerating the flow speed of the liquid and improving the efficiency. In one embodiment, the center line of the port 111 is inclined toward the through hole 134. It can be understood that in this embodiment, the center line of the port 111 is inclined toward the through hole 134, which means that the center line of the first opening is inclined toward the first flow hole, and the center line of the second opening is inclined toward the second flow hole. In this way, when the liquid (for example, cell fluid) is injected into the first containing groove 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 containing groove 121 through the first flow hole and the second flow hole more quickly, thereby reducing the residual cell fluid.

[0038] In one embodiment, the culture dish 100 further includes a filter membrane installed in the port 111. It can be understood that the filter membrane includes but is not limited to a sterile filter membrane, an air filter membrane, etc. The provision of the filter membrane prevents bacteria, impurities, etc. in the external environment from entering the culture dish 100. When the port 111 is located in the middle of the top cover 110, it can be prevented that when the culture dish 100 rotates, the filter membrane in the port 111 is immersed in the liquid in the culture dish 100, thereby affecting the filtering 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 property of the hydrophobic coating facilitates 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 in the direction from the top cover 110 toward the bottom shell 120. It can be understood that the cross-sectional area of ​​the microporous groove 140 gradually decreases, that is, the microporous groove 140 gradually tilts downward and is accommodated, so that the cells in the cell fluid (cell density is greater than the cell fluid) can fall to the bottom of the microporous groove 140 without additional centrifugal force.

[0041] In one embodiment, the microporous groove 140 is a conical groove. It is understandable that the microporous groove 140 can be a quadrangular pyramid groove or a hexagonal pyramid groove, etc. In this embodiment, the top of the microporous groove 140 is a sharp edge, which can prevent the cells from settling and differentiating at the edge of the hole of the microporous groove 140, and the microporous groove 140 has a simple structure and low manufacturing cost. In another specific embodiment, the microporous groove 140 is a conical groove. It is understandable that the inner wall of the microporous groove 140 is a smooth curved surface, which can be more conducive to the agglomeration and growth of cells.

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

[0043] In one embodiment, the slot area of ​​the microporous slot 140 is 0.1 mm. It can be understood that the cross-sectional area of ​​the top of the microporous slot 140 can be set to 0.1 mm according to actual needs. 2 , 0.2mm 2 , 0.3mm 2 , 0.4mm 2 or 0.5mm 2 wait.

[0044] like Figure 5 As shown, an embodiment of the present invention further provides a cell culture method, which is performed by the cell culture device, and the cell culture method comprises:

[0045] S10, injecting liquid into the first receiving groove 131 and / or the second receiving groove 121 of the culture dish 100. It can be understood that the culture dish 100 is provided with an opening for injecting liquid (such as cell fluid or nutrient solution, etc.), and the liquid can be injected into the first receiving groove 131 and / or the second receiving groove 121 through the opening. The above-mentioned opening can be provided on the top cover 110, the intermediate shell 130 or the bottom shell 120. In one embodiment, the above-mentioned opening refers to the port 111 provided on the top cover 110 and connected to the first receiving groove 131, and the liquid (such as cell fluid or nutrient solution, etc.) can be injected into the first receiving groove 131 and / or the second receiving groove 121 through the port 111 on the top cover 110.

[0046] S20, controlling the driving mechanism 210 to drive the culture dish 100 to perform a first motion operation, so that the injected liquid is evenly distributed to all the first containing grooves 131 and the second containing grooves 121. The first motion operation includes but is not limited to rotation, and may also include up, down, left, and right translation.

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

[0048] The driving mechanism 210 is controlled to drive the culture dish 100 to rotate toward the 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 driving mechanism 210 is controlled to drive the culture dish 100 to rotate so that the through hole 134 of the culture dish 100 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; at this time, the driving mechanism 210 is controlled to drive the culture dish 100 to rotate, so that the first receiving groove 131 and the second receiving groove 121 of the culture dish 100 are in a vertical state. At this time, the liquid in the first containing groove 131 and the second containing groove 121 will flow to the same horizontal plane through the through hole 134 , and the liquid will be evenly distributed in the first containing groove 131 and the second containing groove 121 .

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

[0050] The driving mechanism 210 is controlled to drive the culture dish 100 to rotate toward the second direction (i.e., the rotation direction in which the first flow hole and the second flow hole can be located at the bottom), so that the first flow hole and the second flow hole are both located at the bottom of the culture dish 100. It can be understood that the first flow hole and the second flow hole are respectively located at two adjacent corners of the first culture plate 132, so that when the driving mechanism 210 drives the culture dish 100 to rotate to a vertical state, the first flow hole and the second flow hole can be rotated to the bottom of the culture dish 100 at the same time, and then, the liquid in the first containing groove 131 and the second containing groove 121 can be connected through the first flow hole and the second flow hole at the same time, which can speed up the flow speed of the liquid and improve efficiency.

[0051] Wherein, a liquid level sensor may be used to detect whether the liquids in the first containing tank 131 and the second containing tank 121 are at the same level.

[0052] S30, controlling the driving 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 located 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 of the microporous grooves 140, and the amount of liquid in all of the microporous grooves 140 is equal. The second motion operation includes but is not limited to rotation. Specifically, step S30 includes:

[0053] The step S30 comprises:

[0054] S31, after the liquid in the first receiving groove 131 and the second receiving groove 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 toward a third direction (understandably, the third direction should be opposite to the rotation direction in step S20, for example, if the driving mechanism 210 is controlled to drive the culture dish 100 to rotate toward the first direction in step S20, then the third direction should be opposite to the first direction; and if the driving mechanism 210 is controlled to drive the culture dish 100 to rotate toward the second direction in step S20, 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. 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 in the first containing groove 131 and the second containing groove 121 will flow into all the microporous grooves 140 under the action of gravity.

[0055] In step S20, after the liquid in the first containing groove 131 and the second containing groove 121 of the culture dish 100 flows to the same horizontal plane through the through hole 134, the driving mechanism 210 will be further controlled in step S30 to drive the culture dish 100 to rotate, 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 located 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 containing groove 131 and the second containing groove 121 will flow into all the microporous grooves 140 under the action of gravity, and the amount of liquid in all the microporous grooves 140 is equal. Among them, whether the liquid in the microporous grooves 140 is in the same horizontal plane can be detected by a liquid level sensor.

[0056] S32, controlling the driving mechanism 210 to drive the culture dish 100 to move back and forth, so that the liquid adhering to the top cover 110 or the bottom of the first receiving groove 131 is accelerated to drip (drip into the first receiving groove 131 located below the top cover 110, or another first receiving groove 131 located below the first receiving groove 131 or the second receiving groove 121). That is, since the liquid (such as cell fluid, etc.) is viscous, when the top cover 110 of the culture dish 100 is located above the bottom shell 120, a small amount of liquid may still adhere to the top cover 110 or the bottom of the first receiving groove 131. At this time, the driving mechanism 210 is controlled to drive the culture dish 100 to swing back and forth in the horizontal direction. While preventing the liquid in the first receiving groove 131 from flowing from the through hole 134 to the second receiving groove 121, the liquid at the bottom of the first receiving groove 131 can be accelerated to drip into the first receiving groove 131 or the second receiving groove 121 below in the reciprocating swinging motion state, thereby improving efficiency.

[0057] It can be understood that the above cell culture method is not limited to that described in the above embodiments. The specific configuration of the cell culture method of the present invention corresponds to the above cell culture device one by one, and will not be described in detail here.

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

[0059] The specific configuration of the controller of the present invention corresponds one-to-one to the above-mentioned cell culture method, and will not be repeated here. The controller is connected to the driving mechanism 210 of the above-mentioned cell culture device, and then the controller can control the driving 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 microporous grooves 140. Each module in the above-mentioned controller can be implemented in whole or in part by software, hardware and a combination thereof. The above-mentioned modules can be embedded in or independent of the controller in the form of hardware, or can be stored in the controller in the form of software, so that the controller can call and execute the operations corresponding to the above-mentioned modules.

[0060] An embodiment of the present invention further provides a computer-readable storage medium, wherein the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the above-mentioned cell culture method is implemented.

[0061] Those skilled in the art can understand that all or part of the processes in the above-mentioned embodiments can be implemented by instructing related hardware through computer-readable instructions. The computer-readable instructions can be stored in a non-volatile readable storage medium or a volatile readable storage medium. When the computer-readable instructions are executed, they can include the processes of the embodiments of the above-mentioned methods. Among them, any reference to memory, storage, database or other media used in the embodiments provided by the present invention may 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 many forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDRSDRAM), enhanced SDRAM (ESDRAM), synchronous link (Synchlink) DRAM (SLDRAM), memory bus (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 of description, only the division of the above-mentioned functional units and modules is used as an example. In actual applications, the above-mentioned functions can be distributed and completed by 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 device, 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 in the protection scope of the present invention.

Claims

1. A cell culture device, characterized in that: Including a culture dish and a mixing device; The culture dish comprises a top cover, a bottom shell, and at least one intermediate shell stacked and connected between the top cover and the bottom shell, wherein the intermediate shell is recessed on one side facing the top cover to form a first receiving groove, and adjacent first receiving grooves are connected to each other; the bottom shell is recessed on one side close to the intermediate shell to form a second receiving groove, and the second receiving groove is connected to the first receiving groove adjacent thereto; The bottom of the first containing tank and the bottom of the second containing tank are both provided with a plurality of microporous grooves for culturing cells at intervals; The mixing device comprises 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 microporous grooves.

2. The cell culture device according to claim 1, characterized in that: The mixing device comprises a base; the driving mechanism comprises a sliding rod, a telescopic rod and a clamping member 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 a 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 is used to drive the telescopic member, the clamping member and the culture dish to translate along a third reference line, and the third reference line is perpendicular to both the first reference line and the second reference line.

3. The cell culture device according to claim 1, characterized in that: The intermediate shell includes a first culture plate and a first outer frame arranged around the first culture plate; the first containing groove is surrounded by the first outer frame and the first culture plate; the bottom shell includes a second culture plate and a second outer frame arranged around the second culture plate; the second containing groove is surrounded by the second outer frame and the second culture plate; the microporous groove is arranged on the first culture plate and the second culture plate; The first culture plate and / or the first outer frame is provided with through holes; adjacent first containing grooves, and the first containing grooves and the second containing grooves are connected to each other through the through holes.

4. The cell culture device according to claim 3, characterized in that: The top cover is provided with a port communicating with the first containing groove; the port is located in the middle of the top cover.

5. The cell culture device according to claim 3, characterized in that: The top cover is provided with a port communicating with the first accommodating groove; the top cover is rectangular, and the port includes a first opening and a second opening, and the first opening and the second opening are respectively located at two adjacent corners of the rectangular top cover.

6. The cell culture device according to claim 4 or 5, characterized in that: The center line of the port is inclined toward the through hole.

7. The cell culture device according to claim 4 or 5, characterized in that: The culture dish also includes a filter membrane installed in the port.

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

9. A cell culture method, characterized in that: The cell culture method is performed by the cell culture device according to any one of claims 1 to 8, and the cell culture method comprises: Injecting liquid into the first containing tank and / or the second containing tank of the culture dish; Controlling the driving mechanism to drive the culture dish to perform a first movement operation so that the injected liquid is evenly distributed to all the first containing grooves and the second containing grooves; The driving mechanism is controlled to drive the culture dish to perform a second movement 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 containing grooves and the second containing grooves flows into all the microporous grooves, and the amount of liquid in all the microporous grooves is equal.

10. A controller, characterized in that: The controller includes a processor and a memory, wherein the memory stores an executable program, and the processor is used to execute the executable program to implement the cell centrifugation method according to claim 8.

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