Temperature-controlled culture tank for multi-parameter collaborative optimization of CHO (Chinese hamster ovary) cell culture

Abstract

The invention belongs to the technical field of CHO cell culture, and particularly relates to a multi-parameter collaborative optimization temperature control culture tank. The culture tank comprises a tank body, a stirring assembly, a ventilation assembly and a controller, the stirring assembly comprises a vertically-arranged main stirring paddle and a transversely-arranged auxiliary stirring paddle, the main stirring paddle and the auxiliary stirring paddle are each provided with a plurality of stirring blades with different lengths, and the stirring blades are matched with the arc-shaped inner wall of the tank body; the main stirring paddle can break bubbles to increase the oxygen dissolving rate, and the auxiliary stirring paddle can form uniform axial fluid in the tank, reduce cell clusters and nutrition gradient, reduce shear force, realize efficient mass and oxygen transfer and avoid damage to shear force sensitive CHO cells; and a central controller and various sensors are matched, so that the stirring rotating speed and the introduction proportion and rate of each gas can be synergistically regulated and controlled, the culture environment is dynamically maintained to be in an optimal state, and the efficient culture of CHO cells is assisted.

Description

Technical Field

[0001] This invention belongs to the field of CHO cell culture technology, specifically relating to a temperature-controlled culture vessel for CHO cell culture that optimizes multiple parameters in a synergistic manner. Background Technology

[0002] Mammalian cell culture technology is a core pillar of modern biopharmaceuticals and an indispensable technology for the industrial production of key biological products such as recombinant protein drugs, various vaccines, and therapeutic antibodies. Among these, Chinese hamster ovary cells (CHO cells), with their unique biological advantages, have become the preferred cell line for large-scale production in this field. CHO cells possess the ability to precisely fold complex proteins and perform necessary post-translational modifications. This core characteristic directly ensures the structural integrity and biological activity of recombinant biological products, providing crucial support for their clinical application value.

[0003] In the large-scale culture of CHO cells, precise control of key physicochemical parameters such as temperature, pH, and dissolved oxygen concentration is crucial. These parameters not only play a decisive regulatory role in the proliferation activity, cell cycle distribution, and metabolic homeostasis of CHO cells, but also directly affect core quality attributes such as the expression level, purity, and biological activity of target proteins. Practice has shown that even a slight deviation in any parameter can lead to cell growth arrest, metabolic pathway disorders, or even misfolding or abnormal modification of target proteins, thus losing their clinical application value.

[0004] While current commercial bioreactors possess basic cell culture capabilities, significant technological bottlenecks remain for large-scale applications, the most prominent being the core contradiction between mixing efficiency and shear force. This contradiction stems from the design rationality of the stirring system: insufficient stirring intensity leads to concentration gradients and oxygen transfer blind spots within the reactor, resulting in low mass transfer efficiency and an inability to meet the growth and metabolic needs of high-density cell populations; conversely, excessive stirring intensity generates excessive shear force, causing irreversible damage to the cell membrane and increased apoptosis rate in shear-sensitive CHO cells, thereby significantly inhibiting cell proliferation and the efficient expression of target proteins.

[0005] In summary, how to regulate and coordinate the mixing intensity and shear force in the culture vessel to increase cell proliferation and efficient expression of target proteins has become an urgent problem for researchers in this field. Summary of the Invention

[0006] The technical problem to be solved by this invention is: how to achieve the adjustment and coordination of mixing intensity and shear force in the culture tank; To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows: This invention relates to a temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture, comprising: a vessel body; and a stirring assembly, the stirring assembly including a main stirring paddle and an auxiliary stirring paddle, wherein the rotating shaft of the main stirring paddle is vertically disposed at the bottom of the vessel body, the rotating shaft of the auxiliary stirring paddle is horizontally disposed on the vessel body, and the main stirring paddle is located below the auxiliary stirring paddle; the main stirring paddle includes a connecting rod rotatably connected to the vessel body, one end of the connecting rod located inside the vessel body is fixedly connected to a hollow shaft, and two adjustable impellers are fixedly connected to the outside of the hollow shaft, each adjustable impeller including a connecting part fixedly connected to the outside of the hollow shaft, and multiple rotating rods rotatably connected to the periphery of the connecting part, one end of each of the multiple rotating rods being fixedly connected to an impeller body; an angle adjustment mechanism is fixedly connected inside the hollow shaft, and each rotating rod is drivenly connected to the angle adjustment mechanism, the angle adjustment mechanism rotating the impeller body around the axis of the rotating rod body.

[0007] Furthermore, the angle adjustment mechanism includes a core rod rotatably connected inside the hollow shaft. Two first bevel gears are fixedly connected to the outer side of the core rod, and a second bevel gear is fixedly connected to one end of the rotating rod body located inside the hollow shaft. The first and second bevel gears are meshed together. A self-locking drive mechanism that is driven by the core rod is also fixedly connected inside the hollow shaft.

[0008] Furthermore, the self-locking drive mechanism includes a servo motor, a worm gear body, and a worm wheel body. The worm wheel body is fixedly connected to the outside of the core rod, the servo motor is fixedly connected to the inside of the hollow shaft, a first auxiliary bracket is fixedly connected to the inside of the hollow shaft, the worm gear body is rotatably connected to the first auxiliary bracket, and the worm gear body and the worm wheel body are meshed together. The output end of the servo motor is fixedly connected to the worm gear body.

[0009] Furthermore, the hollow shaft includes two cylindrical bodies, and the two connecting parts are respectively fixedly connected to the outside of the two cylindrical bodies. One of the cylindrical bodies is fixedly connected to the connecting rod, and a connecting pipe is fixedly connected to the cylindrical body. The connecting pipe is slidably connected to the other cylindrical body. The core rod includes two inner rotating rods. A second auxiliary support is fixedly connected inside each of the two cylindrical bodies. The two inner rotating rods are rotatably connected to the second auxiliary supports inside the two cylindrical bodies. Two first bevel gears are fixedly connected to the outer sides of the two inner rotating rods. One inner rotating rod is fixedly connected to a worm gear. A sleeve and a prism are fixedly connected to the ends of the two inner rotating rods that are close to each other. The sleeve and the prism are slidably connected. An electric telescopic rod is fixedly connected to the second auxiliary support inside one of the cylindrical bodies. The piston rod of the electric telescopic rod is fixedly connected to the second auxiliary support inside the other cylindrical body.

[0010] Furthermore, it also includes: a sensor assembly, which includes a temperature sensor, a pH sensor, and a dissolved oxygen sensor disposed inside the tank; Ventilation assembly, the ventilation assembly including oxygen manifold, air manifold and C Branch pipes, including the oxygen branch pipe, air branch pipe, and C... Each branch pipe is equipped with a solenoid valve, and The central controller, in which the temperature sensor, pH sensor, dissolved oxygen sensor and each solenoid valve are electrically connected, is configured to coordinately control the rotation speed of the stirring assembly and the gas introduction ratio and rate of the aeration assembly based on sensor data.

[0011] Furthermore, it also includes a temperature control component, which includes a jacketed water bath layer disposed on the outer wall of the tank and at least two temperature sensors disposed at different heights inside the tank.

[0012] Furthermore, the main stirring impeller is a radial flow turbine impeller, and the auxiliary stirring impeller is a slanted blade impeller or an airfoil impeller.

[0013] Furthermore, it also includes a first motor and a second motor. The first motor drives the main stirring paddle to rotate, and the second motor drives the auxiliary stirring paddle to rotate. The first motor and the second motor are electrically connected to the central controller, and their speeds are controlled by the central controller. A tank cover is detachably connected to the tank body via a pin, and each branch pipe of the ventilation assembly is installed on the tank cover.

[0014] Furthermore, the main stirring impeller includes two layers of radial flow turbine impellers, and the auxiliary stirring impeller includes multiple layers of oblique blades arranged in parallel.

[0015] Furthermore, the height-to-diameter ratio of the tank body is between 1.5:1 and 2:1.

[0016] The beneficial effects of this invention are as follows: This invention is a multi-parameter synergistic optimization temperature-controlled culture vessel for CHO cell culture. Through innovative stirring system design and intelligent control scheme, it effectively solves the core contradiction between mass and oxygen transfer and shear damage in traditional culture equipment. The device uses a vertically arranged main stirring paddle, which can efficiently break up air bubbles generated during the culture process and significantly improve the dissolved oxygen transfer rate in the culture medium. At the same time, it is combined with a horizontally arranged auxiliary stirring paddle, which can create a uniform axial fluid circulation in the vessel. On the one hand, it reduces CHO cell aggregation, and on the other hand, it eliminates the concentration gradient of nutrients and dissolved oxygen in the vessel. More importantly, the dual stirring synergistic design greatly reduces fluid shear force, providing a low-damage culture environment for shear-sensitive CHO cells and ensuring cell viability and proliferation efficiency. In terms of intelligent control, the equipment integrates a central controller and sensor components to achieve coordinated closed-loop control of key culture parameters. Through real-time acquisition of core parameters such as stirring speed, the proportion and flow rate of each gas (such as oxygen and carbon dioxide), and jacket water bath temperature by sensors, the central controller dynamically adjusts each execution unit based on a preset algorithm to keep the key environmental indicators of the culture system, such as dissolved oxygen concentration, pH value, and temperature, in the optimal range for CHO cell growth and target protein expression, providing stable and reliable technical support for large-scale and efficient culture. In terms of structural design, based on the CHO cell growth stage, an angle adjustment mechanism is used to adjust the angle of the leaf pulp during rotation, and an electric telescopic rod is used to adjust the height between the leaf pulps to increase cell proliferation and efficient expression of target proteins. Attached Figure Description

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0018] Figure 1 This is a 3D view of a temperature-controlled incubator; Figure 2 This is a plan view of a temperature-controlled incubator; Figure 3 This is a schematic diagram of the interior of a temperature-controlled incubator; Figure 4 This is an external schematic diagram of the main stirring paddle in Example 2; Figure 5 This is a schematic diagram of the interior of the main stirring paddle in Example 2; Figure 6 yes Figure 5 A magnified view of a section at point A in the middle; Figure 7 This is a structural schematic diagram of the core rod.

[0019] In the diagram: 1. Tank body; 2. Stirring assembly; 3. Ventilation assembly; 4. Central controller; 5. Tank lid; 6. Temperature control assembly; 7. pH sensor; 8. Dissolved oxygen sensor; 9. Frame; 10. First motor; 11. Second motor; 12. First auxiliary support; 13. Connecting rod; 14. Cylindrical body; 15. Connecting pipe; 16. Second auxiliary support; 17. Inner rotating rod; 18. Sleeve body; 19. Ribbed rod body; 20. First bevel gear; 21. Second bevel gear; 22. Rotating rod body; 23. Impeller body; 24. Connecting part; 25. Electric telescopic rod; 26. Servo motor; 27. Worm gear body; 28. Worm wheel body; 201. Main stirring paddle; 202. Auxiliary stirring paddle; 301. Oxygen branch pipe; 302. Air branch pipe; 303. CO2 branch pipe; 601. Temperature sensor; 602. Jacketed water bath. Detailed Implementation

[0020] The present invention will now be described in further detail with reference to the accompanying drawings. These drawings are simplified schematic diagrams, illustrating only the basic structure of the invention, and therefore only show the components relevant to the invention.

[0021] like Figure 1 and 2 As shown, a temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture includes a vessel body 1, a stirring assembly 2, a ventilation assembly 3, and a central controller 4. The inner wall of the vessel body 1 is equipped with a temperature sensor 601, a pH sensor 7, and a dissolved oxygen sensor 8. The stirring assembly 2 includes a main stirring paddle 201 and an auxiliary stirring paddle 202. The shaft of the main stirring paddle 201 is vertically positioned at the bottom of the vessel body 1, and the shaft of the auxiliary stirring paddle 202 is horizontally positioned at the upper part of the vessel body 1. The ventilation assembly 3 includes an oxygen manifold 301, an air manifold 302, and a C… Branch pipe 303, oxygen branch pipe 301, air branch pipe 302 and C Each of the branch pipes 303 is equipped with a solenoid valve. The central controller 4 is used to acquire data from the temperature sensor 601, pH sensor 7 and dissolved oxygen sensor 8 to coordinate the control of the rotation speed of the stirring assembly 2 and the opening degree of each solenoid valve.

[0022] A lid 5 is detachably connected to the tank body 1 via a pin. By installing a temperature sensor 601, a pH sensor 7, and a dissolved oxygen sensor 8 on the inner wall of the tank body 1, key parameters such as temperature, pH, and dissolved oxygen concentration can be monitored in real time, providing a stable environment for cell growth. The stirring assembly 2 combines a main stirring paddle 201 and an auxiliary stirring paddle 202. The main stirring paddle 201 is vertically positioned at the bottom of the tank, providing strong mixing capabilities and ensuring uniform distribution of substances within the tank body 1. The auxiliary stirring paddle 202 is horizontally positioned at the top of the tank, further optimizing fluid dynamics and reducing cell clustering and nutrient gradients. The ventilation assembly 3 includes an oxygen manifold 301, an air manifold 302, and a C... Each of the 303 sub-pipes is equipped with a solenoid valve, which can precisely adjust the gas flow rate and ratio according to the cell growth requirements to achieve good mass and oxygen transfer. The controller is responsible for acquiring sensor data and coordinating the control of the stirring component speed and the opening of each solenoid valve, thereby ensuring the conditions required for cell growth while effectively reducing shear force and avoiding damage to shear-sensitive CHO cells.

[0023] The controller employs a programmable logic controller (PLC), which boasts powerful data processing capabilities and stable control performance. It pre-stores various optimized process programs suitable for CHO cell culture, such as "high-density growth mode" and "protein expression mode," allowing users to select the appropriate mode or customize parameter curves. The PLC can receive real-time data from temperature sensor 601, pH sensor 7, and dissolved oxygen sensor 8, and based on preset programs, collaboratively control the stirring speed, the proportion and rate of each gas introduction, and the jacket water bath temperature to dynamically maintain the culture environment at its optimal state. By comparing and analyzing these data with preset parameters, the PLC will react rapidly if any parameter exceeds the preset range, adjusting the speed of the stirring assembly 2 or the opening of the corresponding solenoid valve. For example, when the dissolved oxygen sensor 8 detects that the dissolved oxygen level in tank 1 is lower than the set value, the programmable logic controller will control the opening of the solenoid valve on the oxygen branch pipe 301 to increase the oxygen flow rate; when the temperature sensor 601 detects that the temperature deviates from the suitable range, the programmable logic controller will adjust the temperature inside tank 1 by adjusting the jacket water bath temperature to maintain it in an environment suitable for CHO cell growth.

[0024] like Figure 3 As shown, the main stirring impeller 201 is located at the bottom of the tank 1, preferably a radial flow turbine impeller, used to break up bubbles and increase the dissolved oxygen rate. The auxiliary stirring impeller 202 is located at the upper part of the tank 1, preferably an oblique blade impeller or an airfoil impeller, used to form a uniform axial fluid in the tank 1, reduce cell clusters and nutrient gradients, and at the same time reduce shear force, achieving good mass and oxygen transfer and avoiding damage to shear-sensitive CHO cells.

[0025] To achieve precise temperature control within the tank 1, a temperature control component 6 is also included. This component comprises a jacketed water bath 602 fitted onto the outer wall of the tank 1 and at least two temperature sensors 601 positioned at different heights inside the tank 1. A frame 9 is provided outside the tank 1, and the jacketed water bath 602 is mounted on the frame 9. The jacketed water bath 602 regulates the temperature of the tank 1 by circulating hot or cold water, ensuring a uniform and stable temperature within the tank. The temperature sensors 601 monitor the temperature at different heights within the tank 1 in real time and transmit the data to a controller. The controller automatically adjusts the water temperature in the jacketed water bath 602 according to a preset temperature range, thereby achieving precise temperature control within the tank 1. Through multi-point monitoring and feedback, precise control of the temperature within the tank 1, especially temperature changes during variable-temperature cultivation, is achieved.

[0026] One end of the rotating shaft of the main stirring paddle 201 and the auxiliary stirring paddle 202 passes through the tank body 1, and a sealing ring is provided between the rotating shaft and the tank body 1. The function of the sealing ring is to maintain the sealing effect of the tank body 1 while ensuring rotation. The frame 9 is equipped with a first motor 10 for driving the main stirring paddle 201 and a second motor 11 for driving the auxiliary stirring paddle 202.

[0027] Preferably, the height-to-diameter ratio of the tank 1 is designed to be between 1.5:1 and 2:1, which can better meet the needs of material mixing and transfer within the bioreactor, while optimizing the uniformity of the culture environment.

[0028] Preferably, the central controller 4 integrates a touchscreen. This screen not only intuitively displays various real-time parameters, such as temperature, pH, dissolved oxygen (DO) concentration, and stirring speed, but also provides a user-friendly human-machine interface for setting and adjusting control strategies. The controller is pre-installed with an intelligent program that automatically and dynamically switches relevant parameters according to different growth stages of CHO cells. For example, when cells enter the stationary phase, the system automatically lowers the culture temperature from the conventional 37°C to between 32-34°C according to preset logic to slow down cell metabolism. The system also fine-tunes the pH and dissolved oxygen setpoints to create more suitable culture conditions, thereby extending the culture period and significantly improving protein expression levels.

[0029] like Figure 4-7 As shown, in this embodiment, the main stirring paddle 201 includes a connecting rod 13, which is rotatably connected to the tank body 1. One end of the connecting rod 13 is connected to the first motor 10. A hollow shaft is fixedly connected to one end of the connecting rod 13 inside the tank body 1. Two adjustable impellers are fixedly connected to the outside of the hollow shaft. The adjustable impellers include a connecting part 24 fixedly connected to the outside of the hollow shaft. Multiple rotating rods 22 are rotatably connected to the periphery of the connecting part 24. One end of each of the multiple rotating rods 22 is fixedly connected to an impeller 23. An angle adjustment mechanism is fixedly connected inside the hollow shaft. The rotating rods 22 are all connected to the angle adjustment mechanism. The angle of the rotating rods 22 can be adjusted through the angle adjustment mechanism, thereby adjusting the angle of the impeller 23. The angle of the impeller 23 can be dynamically adjusted according to temperature, pH, DO, speed, etc.

[0030] Among them, the angle adjustment mechanism can be adjusted independently. The number of adjustment mechanisms is the same as that of the rotating rod 22. The adjustment mechanisms and the rotating rod 22 are one-to-one. By activating the corresponding adjustment mechanism, the corresponding rotating rod 22 can be driven to rotate.

[0031] In other embodiments, the angle adjustment mechanism includes a core rod rotatably connected inside the hollow shaft. Two first bevel gears 20 are fixedly connected to the outside of the core rod. A second bevel gear 21 is fixedly connected to one end of the rotating rod body 22 located inside the hollow shaft. The first bevel gears 20 and the second bevel gears 21 are meshed together. A self-locking drive mechanism that is connected to the core rod is also fixedly connected inside the hollow shaft to drive the core rod to rotate. When the core rod rotates, it can synchronously drive multiple blade bodies 23 to rotate through the first bevel gears 20 and the second bevel gears 21.

[0032] like Figure 6 and 7 As shown, the self-locking drive mechanism includes a servo motor 26, a worm gear 27, and a worm wheel 28. The worm wheel 28 is fixedly connected to the outside of the core rod, and the servo motor 26 is fixedly connected to the inside of the hollow shaft. A first auxiliary bracket 12 is fixedly connected to the inside of the hollow shaft. The worm gear 27 is rotatably connected to the first auxiliary bracket 12, and the worm gear 27 and the worm wheel 28 are meshed together. The output end of the servo motor 26 is fixedly connected to the worm gear 27. By starting the servo motor 26, the worm gear 27 can be driven to rotate, which in turn drives the worm wheel 28 to rotate.

[0033] like Figure 4-7 As shown, in some embodiments, the hollow shaft includes two cylindrical bodies 14, and two connecting parts 24 are respectively fixedly connected to the outside of the two cylindrical bodies 14. One of the cylindrical bodies 14 is fixedly connected to the connecting rod 13, and a connecting pipe 15 is fixedly connected to the cylindrical body 14. The connecting pipe 15 is slidably connected to the other cylindrical body 14, so that the hollow shaft is a telescopic shaft, which drives the adjustable propeller located on the upper side to adjust the height.

[0034] The core rod includes two inner rotating rods 17. A second auxiliary support 16 is fixedly connected inside each of the two cylindrical bodies 14. The two inner rotating rods 17 are rotatably connected to the second auxiliary supports 16 inside the two cylindrical bodies 14. Two first bevel gears 20 are fixedly connected to the outer sides of the two inner rotating rods 17. One inner rotating rod 17 is fixedly connected to a worm gear 28. A sleeve 18 and a prism 19 are fixedly connected to the ends of the two inner rotating rods 17 that are close to each other. The sleeve 18 and the prism 19 are slidably connected. An electric telescopic rod 25 is fixedly connected to the second auxiliary support 16 inside one of the cylindrical bodies 14. The piston rod of the electric telescopic rod 25 is fixedly connected to the second auxiliary support 16 inside the other cylindrical body 14. By activating the electric telescopic rod 25 to move the second auxiliary support 16, the inner rotating rod 17 can be moved. Adjusting the length of the core rod adjusts the distance between the two first bevel gears 20, adapting to the height adjustment of the adjustable propeller.

[0035] Both the electric telescopic rod 25 and the servo motor 26 are electrically connected to the central controller 4, which can automatically switch parameters according to the growth stage of CHO cells to adjust the height and tilt angle of the blade body 23.

[0036] Based on the above-described preferred embodiments of the present invention, and through the foregoing description, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification, but must be determined according to the scope of the claims.

Claims

1. A temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture, characterized in that, include: Tank body (1); The stirring assembly (2) includes a main stirring paddle (201) and an auxiliary stirring paddle (202). The rotating shaft of the main stirring paddle (201) is vertically arranged at the bottom of the tank (1), and the rotating shaft of the auxiliary stirring paddle (202) is horizontally arranged on the tank (1). The main stirring paddle (201) is located below the auxiliary stirring paddle (202). The main stirring paddle (201) includes a connecting rod (13), which is rotatably connected to the tank (1). One end of the connecting rod (13) located inside the tank (1) is fixedly connected to a hollow shaft. Two adjustable impellers are fixedly connected to the outside of the hollow shaft. The adjustable impellers include a connecting part (24) fixedly connected to the outside of the hollow shaft. Multiple rotating rods (22) are rotatably connected to the periphery of the connecting part (24). One end of each of the multiple rotating rods (22) is fixedly connected to an impeller (23). An angle adjustment mechanism is fixedly connected inside the hollow shaft. The rotating rods (22) are all connected to the angle adjustment mechanism. The angle adjustment mechanism rotates the impeller (23) around the axis of the rotating rod (22).

2. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, The angle adjustment mechanism includes a core rod rotatably connected inside the hollow shaft. Two first bevel gears (20) are fixedly connected to the outside of the core rod. A second bevel gear (21) is fixedly connected to one end of the rotating rod body (22) located inside the hollow shaft. The first bevel gears (20) and the second bevel gears (21) are meshed together. A self-locking drive mechanism that is connected to the core rod is also fixedly connected inside the hollow shaft.

3. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 2, characterized in that, The self-locking drive mechanism includes a servo motor (26), a worm gear (27), and a worm wheel (28). The worm wheel (28) is fixedly connected to the outside of the core rod. The servo motor (26) is fixedly connected to the inside of the hollow shaft. A first auxiliary bracket (12) is fixedly connected to the inside of the hollow shaft. The worm gear (27) is rotatably connected to the first auxiliary bracket (12), and the worm gear (27) and the worm wheel (28) are meshed. The output end of the servo motor (26) is fixedly connected to the worm gear (27).

4. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 3, characterized in that, The hollow shaft includes two cylindrical bodies (14), and two connecting parts (24) are fixedly connected to the outside of the two cylindrical bodies (14). One of the cylindrical bodies (14) is fixedly connected to a connecting rod (13), and a connecting pipe (15) is fixedly connected to the cylindrical body (14). The connecting pipe (15) is slidably connected to the other cylindrical body (14). The core rod includes two inner rotating rods (17). The interior of each of the two cylindrical bodies (14) is fixedly connected to a second auxiliary support (16). The two inner rotating rods (17) are rotatably connected to the second auxiliary support (16) inside the two cylindrical bodies (14). The two first bevel gears (20) are fixedly connected to the outside of the two inner rotating rods (17). One of the inner rotating rods (17) is fixedly connected to a worm gear (28). The ends of the two inner rotating rods (17) that are close to each other are fixedly connected to a sleeve (18) and a prism body (19). The sleeve (18) and the prism body (19) are slidably connected. An electric telescopic rod (25) is fixedly connected to the second auxiliary support (16) inside one of the cylindrical bodies (14). The piston rod of the electric telescopic rod (25) is fixedly connected to the second auxiliary support (16) inside the other cylindrical body (14).

5. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, Also includes: The sensor assembly includes a temperature sensor (601), a pH sensor (7), and a dissolved oxygen sensor (8) disposed within the tank (1). Ventilation assembly (3), the ventilation assembly (3) including oxygen manifold (301), air manifold (302) and C Branch pipe (303), the oxygen branch pipe (301), the air branch pipe (302) and C Each branch pipe (303) is equipped with a solenoid valve, and The central controller (4) is electrically connected to the temperature sensor (601), pH sensor (7), dissolved oxygen sensor (8) and each solenoid valve. The central controller (4) is configured to coordinately control the rotation speed of the stirring assembly (2) and the gas introduction ratio and rate of the ventilation assembly (3) based on sensor data.

6. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, It also includes a temperature control component (6), which includes a jacketed water bath layer (602) disposed on the outer wall of the tank (1) and at least two temperature sensors (601) disposed at different heights inside the tank (1).

7. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, The main stirring impeller (201) is a radial flow turbine impeller, and the auxiliary stirring impeller (202) is a slanted blade impeller or an airfoil impeller.

8. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, It also includes a first motor (10) and a second motor (11). The first motor (10) drives the main stirring paddle (201) to rotate, and the second motor (11) drives the auxiliary stirring paddle (202) to rotate. The first motor (10) and the second motor (11) are electrically connected to the central controller (4), and their speed is controlled by the central controller (4). The tank body (1) is detachably connected to the tank cover (5) by a pin, and each branch pipe of the ventilation assembly (3) is installed on the tank cover (5).

9. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, The main stirring impeller (201) includes two layers of radial flow turbine impellers, and the auxiliary stirring impeller (202) includes multiple layers of parallel inclined blade impellers.

10. The temperature-controlled culture vessel for multi-parameter synergistic optimization of CHO cell culture according to claim 1, characterized in that, The height-to-diameter ratio of the tank (1) is between 1.5:1 and 2:1.

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