A cell culture device

Abstract

The present application relates to the technical field of cell culture, in particular to a cell culture device, which comprises a culture box with internal constant temperature function, a rotating disc rotatably arranged in the culture box, a plurality of culture dishes arranged along the circumference of the rotating disc and arranged on the rotating disc, and a culture solution adding mechanism; the culture solution adding mechanism comprises a liquid storage container, a liquid outlet member and a liquid discharge control member; the liquid storage container is arranged on the culture box; the liquid outlet member comprises an outer cylinder and an inner core, the outer cylinder is in communication with the liquid storage container through a liquid discharge pipe, a liquid discharge nozzle is arranged on the outer cylinder, and the inner core is rotatably arranged in the outer cylinder and elastically connected to the outer cylinder through a torsional spring; the culture solution adding mechanism is arranged, when the rotating disc rotates forward, the cells in the culture dishes can be uniformly heated in the culture box; when the rotating disc rotates reversely, the culture solution adding mechanism can automatically add culture solution into the culture dishes in the rotating process, so that the accurate addition of culture solution is realized.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, specifically to a cell culture device. Background Technology

[0002] In laboratories, cell culture is often carried out in incubators. Existing technologies, such as the invention patent with patent application number CN202010632537.5, disclose a stem cell culture device that allows the addition of culture medium to a rotating culture dish without opening the incubator door, providing convenience for adding the culture medium. However, this device may have the following problems in practical use: to accurately add the culture medium to the culture dish, a reasonable method of adding the culture medium needs to be set, but the application does not provide a specific solution on how to accurately add the culture medium to the culture dish. Therefore, to improve the above-mentioned device, this invention proposes a cell culture device. Summary of the Invention

[0003] The purpose of this invention is to overcome the shortcomings of the prior art and provide a cell culture device that can automatically and accurately add culture medium into a rotating culture dish and apply mechanical stimulation to the cells.

[0004] This invention is achieved through the following technical solution: A cell culture apparatus includes an incubator with an internal constant temperature function, a turntable rotatably disposed within the incubator, a plurality of culture dishes arranged circumferentially on the turntable, and a culture medium adding mechanism; the culture medium adding mechanism includes a storage container, a dispensing component, and a draining control component; wherein, the storage container is disposed on the incubator; the dispensing component includes an outer cylinder and an inner core, the outer cylinder being connected to the storage container via a drain pipe, the outer cylinder being provided with a drain nozzle, the inner core being rotatably disposed inside the outer cylinder, the inner core being elastically connected to the outer cylinder via a torsion spring, and the inner core having a radially penetrating storage hole corresponding to the positions of the drain nozzle and the drain pipe; the draining control component includes a fixed part, a movable part, and a blocking part, the fixed part being fixed to the inner core, the movable part being connected to the fixed part in a unidirectional rotational manner, and the rotating disk having a blocking part corresponding to the position of each culture dish, which cooperates with the movable part.

[0005] In one implementation, the cell culture device further includes a traction assembly, which includes an elastic membrane, a pull rod, and an impact-receiving component. The elastic membrane is disposed inside the culture dish, and the elastic membrane and the side wall of the culture dish form a culture tank. A first end of the pull rod is connected to the elastic membrane, and a second end of the pull rod abuts against the impact-receiving component. The impact-receiving component is movably disposed on the blocking part, and the side of the impact-receiving component that abuts against the pull rod is an inclined surface.

[0006] In one implementation, a freely rotatable ball is embedded at the end of the second end of the pull rod, and the impacted member makes rolling contact with the ball.

[0007] In one implementation, the turntable is provided with a mounting hole, and the bottom of the culture dish has a flange that is positioned and matched with the mounting hole. The flange has an opening through which the first end of the pull rod can move and pass, and the first end of the pull rod is magnetically fixed to the elastic membrane.

[0008] In one implementation, the flange and the mounting hole are connected by a thread.

[0009] In one implementation, the traction assembly further includes a corrugated inner wall, a piston head, a compression spring, and a piston rod. The corrugated inner wall is substantially cylindrical and disposed inside the incubator. A piston groove is formed on the turntable. The first end of the piston groove communicates with the mounting groove, and the second end of the piston groove extends to the side of the turntable. The piston head is movably disposed within the piston groove. The compression spring is disposed on the side of the piston head facing the mounting groove, and the piston rod is fixed on the other side of the piston head. The piston rod extends out from the second end of the movable groove and abuts against the corrugated inner wall.

[0010] In one implementation, the corrugated inner wall divides the interior of the incubator into a culture chamber and a heating chamber arranged around the culture chamber, with the turntable disposed within the culture chamber.

[0011] In one implementation, the piston rod makes rolling contact with the corrugated inner wall via a roller.

[0012] In one implementation, the outer cylinder is provided with an air inlet, and the line connecting the air inlet and the drain nozzle passes through the axis of the inner core.

[0013] In one implementation, the incubator has an operation window on its top, an openable and closable cover plate at the operation window, ventilation holes on the cover plate, and a filter screen inside the ventilation holes.

[0014] Compared with the prior art, the present invention provides a cell culture device with the following beneficial effects: This invention features a culture medium addition mechanism. When the turntable rotates in the forward direction, the cells in the culture dish are heated evenly in the incubator. When the turntable rotates in the reverse direction, the culture medium addition mechanism automatically adds culture medium to the culture dish during the rotation process, thereby achieving accurate addition of culture medium. This invention, by setting up a traction component that works in conjunction with the drainage control component, can easily apply mechanical stimulation to cells, thereby meeting the experimental needs for studying the effect of mechanical stimulation on the physiological functions of cells. This invention utilizes the combined use of a corrugated inner wall, piston groove, piston head, compression spring, and piston rod to cause the elastic membrane to undergo tensile deformation under the action of air pressure difference during the rotation of the turntable, thereby further applying mechanical stimulation to the cells on the elastic membrane. This invention features a wavy inner wall that can not only work with the compression spring, piston rod, and piston head to apply mechanical stimulation to the cells on the elastic membrane, but also increase the heat conduction area and improve the heating efficiency of the culture chamber. Attached Figure Description

[0015] Figure 1 A three-dimensional structural diagram of a cell culture device; Figure 2 A schematic diagram of the cross-sectional structure of a cell culture device; Figure 3 This is a structural diagram of the turntable and the traction assembly; Figure 4 for Figure 3 A magnified structural diagram at point A; Figure 5 A partial structural diagram of the culture medium addition mechanism; Figure 6 for Figure 5 A schematic diagram of a partial cross-section structure; Figure 7 This is a structural diagram of the turntable, blocking part, pull rod, and impact-receiving part.

[0016] In the diagram: 100, incubator; 101, culture chamber; 102, heating chamber; 110, cover plate; 111, vent hole; 112, filter screen; 200, turntable; 201, mounting hole; 202, piston groove; 210, motor; 300, petri dish; 310, culture tank; 330, flange; 400, liquid storage container; 410, outer cylinder; 411, drain nozzle; 412, air inlet; 420, inner core; 421, liquid storage hole; 421a, liquid inlet; 421b. 430. Liquid outlet; 440. Torsion spring; 440. Fixing part; 440a. Connecting end; 440b. Restricting end; 450. Moving part; 460. Blocking part; 470. Drain pipe; 480. Rotating shaft; 510. Elastic membrane; 511. Pull rod; 512. Ball bearing; 513. Impacted part; 514. Inclined surface; 515. First permanent magnet; 516. Second permanent magnet; 520. Corrugated inner wall; 521. Piston head; 522. Compression spring; 523. Piston rod; 524. Roller. Detailed Implementation

[0017] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] As described in the background section, a stem cell culture device is disclosed in the relevant prior art (patent application number CN202010632537.5). This device allows for the addition of culture medium to a rotating culture dish without opening the door, thus facilitating the addition of culture medium. However, in practical use, this device may have the following problems: to accurately add the culture medium to the culture dish, a reasonable method for adding the culture medium needs to be set, but this application does not provide a specific solution for how to accurately add the culture medium to the culture dish.

[0019] To improve the above device. Example: Please refer to... Figures 1 to 7This invention provides a cell culture apparatus, which mainly includes a culture chamber 100 with internal temperature control, a turntable 200 rotatably disposed within the culture chamber 100, a plurality of culture dishes 300 arranged circumferentially on the turntable 200, and a culture medium adding mechanism. The culture chamber 100 may have a columnar box structure. The culture chamber 100 provides a fully enclosed, independent temperature-controlled space for cell culture. The temperature control effect can be achieved by setting a temperature-controlled heating device inside the culture chamber 100. The temperature-controlled heating device mainly includes an electric heater, a temperature sensor, and a PLC controller. The turntable 200 can rotate in both directions. The rotation of the turntable 200 can be driven by a motor 210. For example, the stator of the motor 210 can be fixedly installed at the bottom of the culture chamber 100, and the rotor of the motor 210 can be coaxially fixed with the turntable 200. The culture dish 300 can be used to hold the cells to be cultured and the culture medium necessary for cell culture. When the motor 210 drives the turntable 200 to rotate, the culture dish 300 can move within the incubator 100, thereby ensuring that the cells within the culture dish 300 are heated evenly within the incubator 100. The culture medium addition mechanism includes a storage container 400, a dispensing component, and a drainage control component. The storage container 400 is located on the incubator 100, specifically on the top of the incubator 100, and is mainly used to hold the culture medium. The dispensing component includes an outer cylinder 410 and an inner core 420. The outer cylinder 410 is connected to the storage container via a drainage pipe 470. The container 400 is connected, and the culture medium in the storage container 400 can be discharged into the outer cylinder 410 through the drain pipe 470. The outer cylinder 410 is provided with a drain nozzle 411, which is used to discharge the culture medium in the outer cylinder 410. The inner core 420 is rotatably disposed inside the outer cylinder 410. The outer wall of the inner core 420 and the inner wall of the outer cylinder 410 are dynamically sealed together. The inner core 420 is elastically connected to the outer cylinder 410 through a torsion spring 430. The inner core 420 is provided with a storage hole 421 in the radial direction, which corresponds to the position of the drain nozzle 411 and the drain pipe 470. One end of the storage hole 421 is the inlet 421a, and the other end is the outlet 421b. The inner core 420 has a first position and a second position relative to the outer cylinder 410. By rotating the inner core 420, it can be switched between these two positions. When the inner core 420 is in the first position, the inlet 421a of the storage hole 421 is connected to the drain pipe 470, and the outlet 421b of the storage hole 421 is sealed by the inner wall of the outer cylinder 410. The culture medium in the storage container 400 can be drained into the storage hole 421 through the drain pipe 470 and temporarily stored in the storage hole 421. When the inner core 420 is in the second position, the inlet 421a of the storage hole 421 is separated from the drain pipe 470, and the outlet 421b of the storage hole 421 is connected to the drain nozzle 411. The culture medium temporarily stored in the storage hole 421 can then be drained out through the drain nozzle 411.The main function of the torsion spring 430 is to provide elastic force for the inner core 420 to return from the second position to the first position. The drainage control component includes a fixed part 440, a movable part 450, and a blocking part 460. The fixed part 440 is fixed to the inner core 420, and the movable part 450 is connected to the fixed part 440 in a unidirectional rotational manner. The rotating disk 200 has a blocking part 460 corresponding to the position of each culture dish 300, which cooperates with the movable part 450. For example, Figure 6 As shown, the inner core 420 is a columnar structure with a large diameter in the middle and a small diameter at both ends. The middle part of the inner core 420 is covered by the outer cylinder 410. The two ends of the inner core 420 extend from the two ends of the outer cylinder 410. The fixed part 440 has a connecting end 440a and a limiting end 440b. The connecting end 440a of the fixed part is fixed to one end of the inner core 420 that extends from the outer cylinder 410. The movable part 450 is rotatably connected to one side of the fixed part 440 through a rotating shaft 480. Under the restriction of the limiting end 440b of the fixed part 440, the movable part 450 can only rotate to one side (i.e., the side away from the limiting end 440b of the fixed part 440).

[0020] Using the above scheme, during cell culture, when no culture medium needs to be added to the culture dish 300, the turntable 200 rotates in the forward direction, causing the culture dish 300 to rotate synchronously, ensuring that the cells in the culture dish 300 are heated evenly and that the culture temperature is consistent; when the blocking part 460 on the turntable 200 encounters the movable part 450, because the movable part 450 is connected to the fixed part 440 in a unidirectional rotation manner, the movable part 450 will flip to allow the blocking part 460 to pass. During this process, the fixed part 440 and the inner core 420 remain stationary, and the inner core 420 is in the first position; when When culture medium needs to be added to the culture dish 300, the turntable 200 rotates in the opposite direction, and the blocking part 460 pushes the moving part 450, the fixed part 440 and the inner core 420 to rotate in the same direction, so that the inner core 420 rotates from the first position to the second position. When the inner core 420 reaches the second position, the culture dish 300 corresponding to the blocking part 460 is exactly below the drain nozzle 411. The outlet 421b of the storage hole 421 is connected to the drain nozzle 411, and the culture medium temporarily stored in the storage hole 421 is accurately drained into the culture dish 300 below through the drain nozzle 411, so as to realize the accurate addition of culture medium.

[0021] In experiments studying the effects of mechanical stimulation on cellular physiological functions, it is necessary to apply mechanical stimulation to the cells. However, the aforementioned approach does not provide a convenient way to perform mechanical stimulation on the cells. Therefore, such as... Figure 3 , Figure 4 and Figure 7As shown, in some embodiments, the cell culture apparatus further includes a traction assembly, which includes an elastic membrane 510 (such as a silicone membrane), a pull rod 511, and a strike-receiving member 513. The elastic membrane 510 is disposed inside the culture dish 300, and the elastic membrane 510 and the side wall of the culture dish 300 form a culture tank 310. The first end of the pull rod 511 is connected to the elastic membrane 510, and the second end of the pull rod 511 abuts against the strike-receiving member 513. The strike-receiving member 513 is movably disposed on the blocking portion 460, and the side of the strike-receiving member 513 that abuts against the pull rod 511 is an inclined surface 514. Specifically, the blocking portion 460 can be a hollow cylinder, the second end of the pull rod 511 is disposed inside the blocking portion 460, and the strike-receiving member 513 moves through the side wall of the blocking portion 460. The end of the strike-receiving member 513 located inside the blocking portion 460 is approximately wedge-shaped, and the end of the strike-receiving member 513 located outside the blocking portion 460 is L-shaped. By setting up a traction assembly that works in conjunction with the drainage control device, mechanical stimulation of cells can be easily achieved, thereby meeting the experimental needs of studying the effect of mechanical stimulation on the physiological functions of cells. In specific operation, cells are placed on the surface of the elastic membrane 510 in the culture tank 310 for culture; during the culture process, the turntable 200 rotates in the forward direction, and the previous blocking part 460 pushes the movable part 450 to flip relative to the fixed part 440; after the previous blocking part 460 passes the movable part 450, the movable part 450 rotates back to its original position under the action of gravity, knocks on the impact member 513 on the adjacent subsequent blocking part 460 and pushes it to move; the impact member 513 drives the pull rod 511 to move through the inclined plane 514, causing the elastic membrane 510 to deform and stretch, thereby applying mechanical stimulation to the cells on the elastic membrane 510.

[0022] like Figure 7 As shown, in some embodiments, a freely rotatable ball 512 is embedded at the second end of the pull rod 511, and the impacted member 513 rolls in contact with the ball 512. This can reduce the contact friction between the pull rod 511 and the impacted member 513, and reduce the motion resistance of the pull rod 511 and the impacted member 513.

[0023] like Figure 4As shown, in some embodiments, the turntable 200 is provided with a mounting hole 201, and the bottom of the culture dish 300 has a protruding flange 330 that positions and engages with the mounting hole 201. To facilitate the assembly and disassembly of the culture dish 300, the flange 330 and the mounting hole 201 can be detachably connected by a thread. The flange 330 has an opening through which the first end of the pull rod 511 can move. The first end of the pull rod 511 is magnetically fixed to the elastic membrane 510. Specifically, a first permanent magnet 515 can be fixedly provided at the first end of the pull rod 511, and a second permanent magnet 516 can be fixedly provided at a corresponding position at the bottom of the elastic membrane 510. The first end of the pull rod 511 and the elastic membrane 510 are magnetically connected through the first permanent magnet 515 and the second permanent magnet 516. This allows the fixing and disassembly operations of the pull rod 511 and the elastic membrane 510 to be completed simultaneously with the assembly and disassembly operations of the culture dish 300.

[0024] like Figure 2 and Figure 4 As shown, in some embodiments, the traction assembly further includes a corrugated inner wall 520, a piston head 521, a compression spring 522, and a piston rod 523. The corrugated inner wall 520 is basically cylindrical and is disposed inside the incubator 100. A piston groove 202 is provided on the turntable 200. The first end of the piston groove 202 is connected to the mounting groove, and the second end of the piston groove 202 extends to the side of the turntable 200. The piston head 521 is movably disposed in the piston groove 202. A compression spring 522 is provided on the side of the piston head 521 facing the mounting groove, and a piston rod 523 is fixed on the other side of the piston head 521. The piston rod 523 extends out from the second end of the movable groove and abuts against the corrugated inner wall 520. By further including the traction assembly with a corrugated inner wall 520, a piston head 521, a compression spring 522, and a piston rod 523, during the rotation of the turntable 200, the piston head 521 and piston rod 523 reciprocate linearly along the piston groove 202 under the elastic force of the compression spring 522 and the guiding push of the corrugated inner wall 520; the reciprocating motion of the piston head 521 will continuously change the air pressure in the mounting groove, thereby causing the elastic membrane 510 to undergo tensile deformation under the action of the air pressure difference, further applying mechanical stimulation to the cells on the elastic membrane 510.

[0025] like Figure 2As shown, in some embodiments, the corrugated inner wall 520 divides the interior of the incubator 100 into a culture chamber 101 and a heating chamber 102 arranged around the culture chamber 101, with a turntable 200 disposed within the culture chamber 101. The heating chamber 102 is used to hold water and is equipped with an electric heater (preferably a resistance heater) for heating the water. When the temperature of the incubator 100 is lower than a set threshold, the electric heater heats the water in the heating chamber 102, and the heat in the water is conducted to the culture chamber 101 through the corrugated inner wall 520, thereby raising the temperature of the internal environment of the culture chamber 101. Furthermore, the electric heater can be linked with a temperature sensor, PLC controller, and other structures to control the water temperature, achieving constant temperature control within the culture chamber 101. It is evident that the corrugated inner wall 520 not only cooperates with the compression spring 522, piston rod 523, and piston head 521 to apply mechanical stimulation to the cells on the elastic membrane 510, but also increases the heat conduction area, improving the heating efficiency of the culture chamber 101.

[0026] like Figure 2 and Figure 4 As shown, in some embodiments, the piston rod 523 rolls against the corrugated inner wall 520 via a roller 524. This effectively reduces the friction between the piston rod 523 and the corrugated inner wall 520, thereby reducing wear at the contact points.

[0027] like Figure 6 As shown, in some embodiments, the outer cylinder 410 is provided with an air inlet 412, and the line connecting the air inlet 412 and the drain nozzle 411 passes through the axis of the inner core 420. By providing an air inlet 412 on the outer cylinder 410, when the inner core 420 is in the second position, the inlet 421a of the storage hole 421 is aligned and connected with the air inlet 412, and the outlet 421b of the storage hole 421 is connected with the drain nozzle 411; the air inlet 412 can maintain the air pressure balance between the inlet 421a and the outlet 421b of the storage hole 421, thereby promoting the rapid discharge of the culture medium in the storage hole 421 and effectively solving the problem of low drainage speed of the storage hole 421.

[0028] like Figure 1 and Figure 2As shown, in some embodiments, the top of the incubator 100 is provided with an operation window, and the operation window is provided with an openable and closable cover 110. The cover 110 is provided with vent holes 111, and a filter screen 112 is provided inside the vent holes 111. Experimenters can take and put the petri dish 300 through the operation window; during the culture process, the cover 110 at the operation window can effectively reduce the temperature loss inside the incubator 100, the vent holes 111 can ensure smooth air circulation inside and outside the incubator 100, and the filter screen 112 filters the air entering the incubator 100. To prevent external pathogens from entering the incubator 100, the filter screen 112 can be made of antibacterial filter material (such as polytetrafluoroethylene nanofiber membrane or HEPA-grade filter material) to further improve the aseptic protection effect.

[0029] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A cell culture device, characterized in that, It includes an incubator (100) with internal constant temperature function, a turntable (200) rotatably disposed inside the incubator (100), a plurality of petri dishes (300) disposed on the turntable (200) and arranged circumferentially on the turntable (200), and a culture medium adding mechanism; The culture medium addition mechanism includes: A liquid storage container (400) is disposed on the incubator (100); The liquid outlet includes an outer cylinder (410) and an inner core (420). The outer cylinder (410) is connected to the liquid storage container (400) via a drain pipe (470). A drain nozzle (411) is provided on the outer cylinder (410). The inner core (420) is rotatably disposed inside the outer cylinder (410) and is elastically connected to the outer cylinder (410) via a torsion spring (430). A liquid storage hole (421) is provided radially through the inner core (420) corresponding to the positions of the drain nozzle (411) and the drain pipe (470). The drain control component includes a fixed part (440), a movable part (450), and a blocking part (460). The fixed part (440) is fixed on the inner core (420), and the movable part (450) is connected to the fixed part (440) in a unidirectional rotation manner. The rotating disk (200) is provided with the blocking part (460) corresponding to the position of each of the culture dishes (300), which cooperates with the movable part (450).

2. The cell culture apparatus according to claim 1, characterized in that: It also includes a traction assembly, which includes an elastic membrane (510), a pull rod (511), and an impact-receiving component (513). The elastic membrane (510) is disposed inside the culture dish (300), and the elastic membrane (510) and the side wall of the culture dish (300) form a culture tank (310). The first end of the pull rod (511) is connected to the elastic membrane (510), and the second end of the pull rod (511) abuts against the impact-receiving component (513). The impact-receiving component (513) is movably disposed on the blocking part (460), and the side of the impact-receiving component (513) that abuts against the pull rod (511) is an inclined surface (514).

3. The cell culture apparatus according to claim 2, characterized in that: The second end of the pull rod (511) is embedded with a freely rotatable ball (512), and the impacted part (513) makes rolling contact with the ball (512).

4. The cell culture apparatus according to claim 2, characterized in that: The turntable (200) is provided with a mounting hole (201), and the bottom of the culture dish (300) has a flange (330) that is positioned and matched with the mounting hole (201). The flange (330) has an opening through which the first end of the pull rod (511) can move through. The first end of the pull rod (511) is magnetically fixed to the elastic membrane (510).

5. The cell culture apparatus according to claim 4, characterized in that: The flange (330) and the mounting hole (201) are connected by a thread.

6. The cell culture apparatus according to claim 4, characterized in that: The traction assembly further includes a corrugated inner wall (520), a piston head (521), a compression spring (522), and a piston rod (523). The corrugated inner wall (520) is basically cylindrical and is disposed inside the incubator (100). A piston groove (202) is provided on the turntable (200). The first end of the piston groove (202) is connected to the mounting groove. The second end of the piston groove (202) extends to the side of the turntable (200). The piston head (521) is movably disposed in the piston groove (202). The compression spring (522) is provided on the side of the piston head (521) facing the mounting groove. The piston rod (523) is fixed on the other side of the piston head (521). The piston rod (523) extends out from the second end of the movable groove and abuts against the corrugated inner wall (520).

7. The cell culture apparatus according to claim 6, characterized in that: The wavy inner wall (520) divides the interior of the incubator (100) into a culture chamber (101) and a heating chamber (102) arranged around the culture chamber (101), and the turntable (200) is disposed in the culture chamber (101).

8. The cell culture apparatus according to claim 6, characterized in that: The piston rod (523) makes rolling contact with the corrugated inner wall (520) via a roller (524).

9. The cell culture apparatus according to claim 1, characterized in that: The outer cylinder (410) is provided with an air inlet (412), and the line connecting the air inlet (412) and the drain nozzle (411) passes through the axis of the inner core (420).

10. The cell culture apparatus according to claim 1, characterized in that: The incubator (100) has an operation window on its top, and an openable and closable cover (110) is provided at the operation window. The cover (110) has a vent (111) and a filter (112) is provided inside the vent (111).

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