AI-based CAR-NK cell culture device and method

Abstract

The invention relates to the technical field of cell culture, in particular to an AI-based CAR-NK cell culture device and method.The device comprises a support and a control system used for controlling operation of the device, the support is rotationally connected with an obliquely-installed straight pipe, and the two ends of the straight pipe are each provided with a connector communicated with the outside; the control system is used for controlling the opening and closing of the electromagnetic valves; the straight pipe is communicated with a spiral pipe, and the support is provided with a driving assembly used for driving the straight pipe to rotate. The driving assembly comprises double output pieces fixedly connected to the bottom of the support, and the control system is used for controlling the double output pieces to rotate. An output shaft of the double-output piece is coaxially and fixedly connected with a main bevel gear, the main bevel gear is meshed with an auxiliary bevel gear, and the auxiliary bevel gear is coaxially and fixedly connected with the straight pipe. Through the synergistic effect of the conveying assembly and the adjusting assembly, the nutrient solution components, the gas concentration and the flow can be flexibly adjusted according to different culture stages.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, and specifically to an AI-based CAR-NK cell culture device and method. Background Technology

[0002] CAR-NK (Chimeric Antigen Receptor Natural Killer) cell therapy is an emerging immunotherapy approach designed to fight diseases such as cancer by enhancing a patient's own immune system. In CAR-NK cell culture, platforms like Ronggu Biotechnology's LEONCAR™ platform cover the entire process from NK cell isolation to lentiviral transduction of the CAR gene, efficient in vitro expansion and production of CAR-NK cells, and programmed freezing for CAR-NK cell preparation.

[0003] While the aforementioned products can culture cells, they lack real-time analysis of environmental parameters. Their regulatory strategies often rely on pre-set programs rather than dynamically responding to changes in actual cellular metabolic activity. The frequency of acquiring biological indicators such as cell growth stages and metabolic needs is low, potentially preventing timely detection and intervention of changes in cell state.

[0004] In summary, how to solve the problem of the lack of real-time analysis of environmental parameters in existing technologies, which may prevent timely detection of changes in cell state and intervention, has become an urgent problem to be solved in this field. Therefore, it is necessary to propose an AI-based CAR-NK cell culture device and method. Summary of the Invention

[0005] To address the aforementioned issues, this invention provides an AI-based CAR-NK cell culture device and method. Through the synergistic effect of delivery and regulation components, the nutrient solution composition, gas concentration, and flow rate can be flexibly adjusted for different culture stages. This regulatory capability not only shortens the culture cycle but also reduces the toxic effects of metabolic waste accumulation on cells, ensuring CAR-NK cells grow in an optimal environment and improving product consistency and safety.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows: an AI-based CAR-NK cell culture device includes a support and a control system for controlling the operation of the device. A straight tube installed at an incline is rotatably connected to the support, and both ends of the straight tube have interfaces for communicating with the outside. Solenoid valves are connected to the interfaces, and the control system is used to control the opening and closing of the solenoid valves. A spiral tube is connected to the straight tube, and a drive component for driving the straight tube to rotate is provided on the support.

[0007] The drive assembly includes a dual output component fixedly connected to the bottom of the bracket. The control system is used to control the rotation of the dual output component. The output shaft of the dual output component is coaxially fixedly connected to a main bevel gear, which meshes with a secondary bevel gear. The secondary bevel gear is coaxially fixedly connected to a straight tube.

[0008] The support is also equipped with a delivery assembly for delivering nutrient solution to the spiral tube and a regulating assembly for regulating the gas environment inside the spiral tube.

[0009] The drive assembly is equipped with a transmission assembly for driving the conveying assembly and the regulating assembly; the transmission assembly is equipped with an opening and closing assembly for driving the conveying assembly and the regulating assembly independently; the control system is used to control the operation of the transmission assembly and the opening and closing assembly respectively to regulate the cell growth environment.

[0010] The technical principles of the above solution are as follows:

[0011] The cells to be cultured are fed into the straight tube by opening the solenoid valve. Since the output shafts of the dual output components are coaxially fixedly connected to a main bevel gear, which meshes with a secondary bevel gear, and the secondary bevel gear is coaxially fixedly connected to the straight tube, the dual output components drive the main bevel gear to rotate, which in turn drives the secondary bevel gear to rotate synchronously, thus driving the straight tube to rotate synchronously. Because the straight tube is connected to a spiral tube, the rotation of the straight tube drives the spiral tube to rotate synchronously. Utilizing the special design of the spiral tube, the cells rise slowly within it, and upon reaching the apex, flow back to the bottom of the straight tube due to gravity, promoting cell adhesion and proliferation. During this process, the delivery and regulating components deliver nutrient solution and required gases to the spiral tube, providing the cells with the necessary environment. The flow rate and composition of the nutrient solution, as well as the gas concentration and flow rate, can be controlled using the opening and closing components and the transmission components according to the cell growth stage and metabolic needs, accelerating cell proliferation and shortening the culture period.

[0012] The above approach has the following beneficial effects:

[0013] 1. This invention utilizes a linked rotational design of a helical tube and a straight tube, allowing cells to continuously contact fresh culture medium during their spiral ascent and naturally settle under gravity, creating a dynamic three-dimensional culture environment. This combination of non-uniform shear force and periodic gravitational stimulation can simulate the in vivo microenvironment, promoting the natural killing activity of NK cells and the stability of CAR structures. Simultaneously, the hydrodynamic effects generated by the helical tube rotation enhance intercellular interactions, thereby improving the anti-tumor efficacy of CAR-NK cells, enhancing cell functional stability, and providing more active cell products for clinical applications.

[0014] 2. This invention, through the synergistic action of the delivery and regulation components, can flexibly adjust the nutrient solution composition, gas concentration, and flow rate for different culture stages. The linked design of the opening / closing and transmission components enables independent control of nutrient solution and gas delivery, reducing the risk of imbalances in other environmental factors due to adjustments of a single parameter. This regulatory capability not only shortens the culture cycle but also reduces the toxic effects of metabolic waste accumulation on cells, ensuring CAR-NK cells grow in an optimal environment and improving product consistency and safety.

[0015] 3. This invention achieves real-time monitoring and dynamic adjustment of culture parameters through the automated integration of the control system and its components. The coordinated control of the solenoid valve and dual output components precisely controls the cell seeding rate and culture medium circulation speed. This automated process effectively reduces manual intervention, thereby lowering the risk of contamination, while standardized operating procedures improve batch-to-batch consistency. Furthermore, the modular design of the device facilitates maintenance and upgrades, further reducing long-term operating costs and providing an economically feasible solution for the large-scale production of CAR-NK cell therapy.

[0016] Furthermore, the conveying assembly includes a first cylinder, a first rod, and a first plate. The first cylinder is fixedly connected to the bottom of the support. The first plate slides vertically against the inner wall of the first cylinder, and the first rod is located above the first plate. The opening and closing assembly is used to connect the first rod and the first plate, and the transmission assembly is used to drive the first rod to reciprocate.

[0017] The bottom of the first cylinder is connected to an input pipe and an output pipe, and the connection between the input pipe and the output pipe is connected to a first one-way valve; the end of the input pipe away from the first cylinder is connected to a storage tank for storing nutrient solution, and the end of the output pipe away from the first cylinder is connected to the inside of a straight pipe.

[0018] Beneficial effects: Precise nutrient solution supply is achieved through mechanical linkage. Driven by a transmission assembly, the first lever pushes the first plate to slide back and forth within the first cylinder, creating a piston pump effect. During upward movement, nutrient solution is drawn in through the input pipe and the first one-way valve; during downward movement, it is pushed unidirectionally into the straight pipe through the output pipe. This mechanical structure, combined with a controllable opening and closing assembly, ensures quantitative delivery of nutrient solution according to cell metabolic needs. Simultaneously, the fully enclosed pipeline design helps reduce the risk of exogenous contamination, meets the aseptic requirements of CAR-NK culture, and guarantees batch stability.

[0019] Furthermore, the adjustment assembly includes a second cylinder, a second rod, and a second plate. The second cylinder is fixedly connected to the side of the bracket away from the first cylinder. The second plate slides vertically against the inner wall of the second cylinder, and the second rod is located above the second plate. The opening and closing assembly is used to connect the second rod and the second plate, and the transmission assembly is used to drive the second rod to reciprocate.

[0020] The bottom of the second cylinder is connected to an inlet pipe and an outlet pipe, and a second one-way valve is connected to the connection between the inlet pipe and the outlet pipe. The end of the inlet pipe away from the second cylinder is connected to a gas storage tank for storing gas, and the end of the outlet pipe away from the second cylinder is connected to the inside of a straight pipe.

[0021] Beneficial effects: Precise gas environment control is achieved through mechanical linkage. Driven by the transmission component, the second rod pushes the second plate to slide back and forth within the second cylinder, forming a piston-like gas pump. When moving upwards, a specific gas mixture is drawn in through the inlet pipe and the second one-way valve; when moving downwards, it is injected unidirectionally into the straight pipe through the outlet pipe. This structure, combined with an on / off controllable mechanism, ensures that the gas concentration and flow rate are dynamically adjusted according to the needs of cell growth stages. Simultaneously, the fully enclosed gas circulation path reduces external contamination, maintains a sterile environment in the culture system, and ensures the functional stability of CAR-NK cells.

[0022] Furthermore, the transmission assembly includes a first gear and a second gear. The first gear is coaxially and fixedly connected to the side of the dual output component away from the main bevel gear. A fixed frame is fixedly connected to the bracket, and the first gear is rotatably connected to the fixed frame. The second gear meshes with the first gear, and an actuating rod is eccentrically and fixedly connected to the second gear. A movable seat is vertically slidably fitted on the fixed frame, and the first rod and the second rod are respectively fixedly connected to both sides of the movable seat. The movable seat has a groove for the movement of the actuating rod. The bracket is also provided with a telescopic assembly for disengaging the first gear and the second gear.

[0023] Beneficial effects: By cooperating with the eccentric actuating rod and the sliding groove, the rotational motion is converted into the smooth reciprocating motion of the moving seat. The second gear drives the actuating rod to slide within the sliding groove of the moving seat. When the actuating rod rotates, its end away from the second gear drives the moving seat to reciprocate, thereby driving the first and second rods to rise and fall synchronously, ensuring controllable frequency of nutrient solution and gas delivery. The device has a high degree of overall structural integration and low transmission loss, which helps to improve the response speed and reliability of environmental parameter adjustment and ensures the dynamic stability of the CAR-NK cell culture process.

[0024] Furthermore, the telescopic assembly includes a telescopic member fixedly connected to the bracket, a control system for controlling the telescopic member to extend and retract, and the output shaft of the telescopic member being rotatably connected to the second gear.

[0025] Beneficial effects: By driving the axial displacement of the telescopic component through the control system, the second gear disengages from or engages with the first gear, directly controlling the power supply to and from the conveying and regulating components. Based on cell growth stages or AI commands, it can cut off or resume power transmission, reducing resource waste or metabolic imbalances caused by ineffective transport.

[0026] Furthermore, the opening and closing assembly includes several electromagnets, and the control system is used to control the opening and closing of the electromagnets; the electromagnets are respectively fixedly connected to the bottom ends of the first rod and the second rod, and magnetic blocks that magnetically cooperate with the electromagnets are embedded inside the first plate and the second plate.

[0027] Beneficial effects: Precise control is achieved through magnetic non-contact transmission. The control system activates and deactivates electromagnets as needed. When energized, the electromagnets attract the magnetic blocks embedded in the first or second plate, causing the rods to rigidly link and perform conveying or regulating actions, synchronously driving the delivery of nutrient solution or gas. When de-energized, the magnetism disappears and the electromagnets automatically separate, cutting off the power transmission. It can independently activate or deactivate nutrient or gas pathways according to AI commands, meeting the environmental parameter regulation requirements for cell culture.

[0028] Furthermore, a gas sensor and a pH sensor are fixedly connected to the inner wall of the spiral tube. The control system is used to receive gas information and pH information emitted by the gas sensor and pH sensor, and to control the opening and closing of the telescopic component and the electromagnet based on the gas information and pH information respectively.

[0029] Beneficial effects: By monitoring key parameters inside the spiral tube through gas and pH sensors, the control system analyzes the data stream in real time and links the actuators. Based on the gas concentration, the expansion joint is started and stopped, and the electromagnet is switched on and off according to pH fluctuations to match the nutrient solution pulse rhythm, ensuring that the cell metabolic microenvironment is in the optimal state. This realizes the change of the culture environment from passive maintenance to active adaptation, providing high-precision physiological support for CAR-NK cell expansion.

[0030] Furthermore, both the straight tube and the spiral tube are made of transparent material, and an image sensor is fixedly connected to the bracket; the control system is used to receive the image information emitted by the image sensor, and control the opening and closing of the telescopic component and the electromagnet based on the image information.

[0031] Beneficial effects: By using image sensors to capture cell morphology and distribution information in real time, the control system analyzes key indicators such as cell aggregation degree and adhesion state through AI vision algorithms, adjusts the gas supply through linkage with telescopic components, and synchronously controls the opening and closing frequency of electromagnets to optimize the nutrient pulse rhythm, thus providing growth protection for CAR-NK cells.

[0032] Furthermore, the control system includes the following modules:

[0033] The data acquisition module is used to collect environmental parameters inside the spiral tube through gas sensors and pH sensors; to collect cell morphology distribution information through image sensors; and to transmit the collected environmental parameters and morphology distribution information to the data analysis module.

[0034] The data analysis module receives data transmitted from the data acquisition module, uses a deep learning model to analyze cell density, activity, and CAR expression rate, and integrates gas information, pH information, and cell morphology distribution information to generate environmental regulation instructions and nutrient supply strategies, and outputs analysis results.

[0035] The execution control module is used to control the start and stop of the telescopic component and electromagnet based on the analysis results of the data analysis module, inject specific mixed gas and deliver nutrient solution as needed, and control the output of the dual output component to adjust the rotation speed of the straight tube and the spiral tube.

[0036] The abnormal calibration module is used to set a threshold for cell growth status based on environmental parameters and morphological distribution information. When the threshold is reached, it controls the opening and closing of the corresponding dual output components, electromagnets and telescopic components, and adjusts the gas injection frequency, nutrient solution supply amount and rotation parameters to maintain the optimal growth status.

[0037] Beneficial Effects: By integrating gas, pH, and image sensors for identification and monitoring, and utilizing a deep learning model to analyze cell metabolic status in real time, a dynamic strategy for generating gas components and nutrient solution pulses is generated. Based on the analysis results, the system synchronously controls the opening and closing of gas pathways via telescopic components, the switching of nutrient supply via electromagnets, and the adjustment of helical tube speed via dual output components, maintaining the optimal balance between cell adhesion and suspension. Furthermore, a growth threshold is set to cut off abnormal supply chains and self-calibrate parameters, reducing the risk of metabolic imbalance. The collaborative design of these modules overcomes the limitations of traditional manual intervention, achieving zero-hysteresis optimization of the culture environment through data-driven approaches, ensuring high activity, high consistency, and suitability for solid tumor treatment in CAR-NK cell expansion.

[0038] Furthermore, an AI-based CAR-NK cell culture method includes the following steps:

[0039] S1. Cell Seeding and Initial Culture: Inject the CAR gene-modified NK cell suspension into the helical tube through the interface at the bottom of the straight tube; start the dual output device to drive the straight tube and helical tube to rotate, and set the initial speed.

[0040] S2. Real-time sensing of multi-source data: The gas information in the spiral tube is monitored in real time by a gas sensor, the pH value of the culture medium is collected by a pH sensor, the cell morphology distribution image is captured by an image sensor, and each monitoring threshold is set.

[0041] S3. Dynamic Decision-Making and Control: If the gas information exceeds the preset parameter threshold, the regulating component will inject mixed gas into the spiral tube; if the pH value exceeds the preset acid-base range, the delivery component will replenish the nutrient solution to adjust the pH value range; when the cell density growth rate is lower than the preset threshold, the rotation speed of the dual output components will be increased and the nutrient solution replenishment frequency will be increased.

[0042] S4. Closed-loop adaptive amplification: Continuously compare the actual cell activity with the preset growth curve. If the actual cell activity exceeds the preset growth curve, adjust the injection frequency of the mixed gas, the amount of nutrient solution supplied at one time, and the rotation period of the spiral tube.

[0043] S5. Harvesting and Quality Control: When the cell density reaches the standard, the solenoid valve is opened to collect the cell suspension; and the killing activity of the cells is verified by flow cytometry to complete batch production.

[0044] Beneficial effects: The automated operation process enables dynamic optimization and expansion of CAR-NK cells. It integrates multi-source data based on gas and environmental information in real time and uses algorithms to intelligently determine environmental regulation thresholds and trigger gas supply, nutrient pulses, and rotation speed adjustment to match cell metabolic rhythms and proliferation needs. It can effectively overcome the bottleneck of metabolic waste accumulation in traditional static culture. The gravity circulation mechanism combined with dynamic parameter domestication maintains high viability and functional stability. Through real-time comparison and adaptive calibration of growth curves, it ensures batch-to-batch consistency and enhances tolerance to the solid tumor microenvironment, providing an efficient and standardized process pathway.

[0045] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0046] Figure 1 This is a side cross-sectional view of the AI-based CAR-NK cell culture device of the present invention.

[0047] Figure 2 This is an isometric view of the installation of the transmission components in the AI-based CAR-NK cell culture device of the present invention.

[0048] Figure 3 This is an isometric view of the installation of the telescopic component in the AI-based CAR-NK cell culture device of the present invention.

[0049] Figure 4 This is a front view of the transmission component in the AI-based CAR-NK cell culture device of the present invention.

[0050] Figure 5 This is a cross-sectional view of the delivery component in the AI-based CAR-NK cell culture device of the present invention.

[0051] Figure 6 This is a cross-sectional view of the regulating component in the AI-based CAR-NK cell culture device of the present invention.

[0052] Figure 7 This is a structural block diagram of the control system in the AI-based CAR-NK cell culture device of the present invention.

[0053] Figure 8 This is a flowchart of the AI-based CAR-NK cell culture method of the present invention.

[0054] The reference numerals in the accompanying drawings of the instruction manual include: 1. bracket; 2. straight tube; 3. spiral tube; 4. dual output component; 5. main bevel gear; 6. secondary bevel gear; 7. first cylinder; 8. first rod; 9. first plate; 10. second cylinder; 11. second rod; 12. second plate; 13. first gear; 14. second gear; 15. fixed frame; 16. actuating rod; 17. movable seat; 18. telescopic component; 19. electromagnet; 20. magnetic block. Detailed Implementation

[0055] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. 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.

[0056] In the description of this invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0057] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0058] The following detailed description illustrates the specific implementation method:

[0059] Example 1:

[0060] As attached Figure 1 As shown: An AI-based CAR-NK cell culture device includes a support 1 and a control system for controlling the operation of the device. A straight tube 2 installed at an angle is rotatably connected to the support 1. Both ends of the straight tube 2 have interfaces for communicating with the outside. Solenoid valves are connected to the interfaces. The control system is used to control the opening and closing of the solenoid valves. A spiral tube 3 is connected to the straight tube 2. The support 1 is provided with a drive component for driving the straight tube 2 to rotate.

[0061] The drive assembly includes a dual output component 4 bolted to the bottom of the bracket 1. In this embodiment, the dual output component 4 is a dual-head motor. The control system is used to control the rotation of the dual output component 4. The output shaft of the dual output component 4 is coaxially fixedly connected to a main bevel gear 5 via a coupling. The main bevel gear 5 meshes with a secondary bevel gear 6. The secondary bevel gear 6 is coaxially bolted to the straight tube 2.

[0062] Combination Figure 2 As shown, the support 1 is also equipped with a delivery component for delivering nutrient solution to the spiral tube 3 and a regulating component for regulating the gas environment inside the spiral tube 3.

[0063] The drive assembly is equipped with a transmission assembly for driving the conveying assembly and the regulating assembly; the transmission assembly is equipped with an opening and closing assembly for driving the conveying assembly and the regulating assembly independently; the control system is used to control the operation of the transmission assembly and the opening and closing assembly respectively to regulate the cell growth environment.

[0064] Combination Figure 5 As shown, the conveying assembly includes a first cylinder 7, a first rod 8, and a first plate 9. The first cylinder 7 is bolted to the bottom of the bracket 1. The first plate 9 is vertically slidably fitted with the inner wall of the first cylinder 7. The first rod 8 is located above the first plate 9. The opening and closing assembly is used to connect the first rod 8 and the first plate 9. The transmission assembly is used to drive the first rod 8 to reciprocate.

[0065] The bottom of the first cylinder 7 is connected to an input pipe and an output pipe. The connection between the input pipe and the output pipe is connected to a first one-way valve. In this embodiment, the first one-way valve is used to guide the unidirectional flow of the medium. The end of the input pipe away from the first cylinder 7 is connected to a storage tank for storing nutrient solution. In this embodiment, the storage tank is used to store the corresponding nutrient solution according to the growth needs of the cells, such as RPMI-1640 culture medium. The end of the output pipe away from the first cylinder 7 is connected to the inside of the straight pipe 2.

[0066] Specifically, when the first rod 8 is driven upward by the transmission assembly, it simultaneously drives the first plate 9 to move upward as well. When the first plate 9 moves upward, a negative pressure is formed in the first cylinder 7, the first one-way valve of the input pipe opens, and the negative pressure draws in the nutrient solution from the storage tank, which then flows into the first cylinder 7 through the input pipe. When the first rod 8 is driven downward by the transmission assembly, the first plate 9 simultaneously presses down, the first one-way valve of the output pipe opens, and the nutrient solution is unidirectionally forced into the straight pipe 2 through the output pipe.

[0067] Combination Figure 6 As shown, the adjustment assembly includes a second cylinder 10, a second rod 11, and a second plate 12. The second cylinder 10 is bolted to the side of the bracket 1 away from the first cylinder 7. The second plate 12 is vertically slidably engaged with the inner wall of the second cylinder 10. The second rod 11 is located above the second plate 12. The opening and closing assembly is used to connect the second rod 11 and the second plate 12. The transmission assembly is used to drive the second rod 11 to reciprocate.

[0068] The bottom of the second cylinder 10 is connected to an inlet pipe and an outlet pipe. A second one-way valve is connected to the connection between the inlet pipe and the outlet pipe. In this embodiment, the second one-way valve is used to guide the unidirectional flow of the medium. The end of the inlet pipe away from the second cylinder 10 is connected to a gas storage tank for storing gas. In this embodiment, the gas storage tank is used to store the gas required by the cells, such as oxygen or carbon dioxide. The end of the outlet pipe away from the second cylinder 10 is connected to the inside of the straight pipe 2.

[0069] Specifically, when the second rod 11 moves upward, the second plate 12 moves upward accordingly, creating a negative pressure inside the second cylinder 10. This negative pressure draws in a specific mixed gas (such as 5% CO2 or a low-oxygen mixed gas) from the gas storage tank, which then flows into the second cylinder 10 through the inlet pipe. When the second rod 11 moves downward, the second plate 12 simultaneously presses down, and the gas is injected unidirectionally into the straight pipe 2 through the outlet pipe, replacing the gas environment inside the spiral tube 3.

[0070] Combination Figure 2 and Figure 4 As shown, the transmission assembly includes a first gear 13 and a second gear 14. The first gear 13 is coaxially and fixedly connected to the side of the double output component 4 away from the main bevel gear 5 via a coupling. A fixed frame 15 is bolted to the bracket 1, and the first gear 13 is rotatably connected to the fixed frame 15. The second gear 14 meshes with the first gear 13, and an actuating rod 16 is eccentrically bolted to the second gear 14. A movable seat 17 is vertically slidably fitted on the fixed frame 15, and the first rod 8 and the second rod 11 are respectively screwed to both sides of the movable seat 17. A sliding groove is opened on the movable seat 17 for the movement of the actuating rod 16.

[0071] Specifically, the dual output components 4 drive the coaxial first gear 13 to rotate, and the first gear 13 meshes with the second gear 14 to rotate synchronously. The eccentric actuating rod 16 of the second gear 14 rotates in a circular motion. The actuating rod 16 slides within the groove of the moving seat 17, at which point the circular motion of the actuating rod 16 is converted into the vertical reciprocating motion of the moving seat 17. Figure 2 For example, when the second gear 14 drives the lever 16 to rotate above it, the lever 16 drives the movable seat 17 to move upward. Conversely, when the lever 16 rotates below the second gear 14, it can cause the movable seat 17 to move downward.

[0072] Combination Figure 3As shown, the bracket 1 is also equipped with a telescopic assembly for disengaging the first gear 13 and the second gear 14. The telescopic assembly includes a telescopic member 18 bolted to the bracket 1. In this embodiment, the telescopic member 18 is an electric actuator. The control system controls the telescopic member 18 to extend and retract. The output shaft of the telescopic member 18 is rotatably connected to the second gear 14. In this embodiment, the rotatable connection between the output shaft of the telescopic member 18 and the second gear 14 enables the second gear 14 to rotate, and allows the output shaft of the telescopic member 18 to drive the second gear 14 to move.

[0073] When the telescopic component 18 is not activated, the second gear 14 remains engaged with the first gear 13, and power is continuously transmitted. When it is necessary to cancel the power transmission, the control system triggers the telescopic component 18 to move its output shaft upward. The telescopic component 18 drives the second gear 14 to move upward, causing the second gear 14 to disengage from the first gear 13, interrupting the power transmission and canceling the delivery of gas and nutrient solution.

[0074] The opening and closing assembly includes several electromagnets 19, and the control system is used to control the opening and closing of the electromagnets 19. The electromagnets 19 are fixedly bonded to the bottom ends of the first rod 8 and the second rod 11 respectively. The first plate 9 and the second plate 12 are both embedded with magnetic blocks 20 that are magnetically matched with the electromagnets 19.

[0075] Specifically, the control system activates electromagnet 19, which generates magnetic attraction with the embedded magnetic blocks 20 in the first plate 9 and the second plate 12, connecting the first rod 8 to the first plate 9 (or the second rod 11 to the second plate 12). When electromagnet 19 is de-energized, the magnetic force disappears, and the first plate 9 and the second plate 12 are no longer actuated. In this embodiment, the on / off frequency of the corresponding electromagnet 19 is intelligently controlled based on the cell growth stage or metabolic needs; the control system activates and deactivates the corresponding electromagnet 19 as needed. When energized, it attracts the embedded magnetic blocks 20 in the first plate 9 or the second plate 12, connecting the first rod 8 to the first plate 9, or the second rod 11 to the second plate 12; nutrient solution or mixed gas is pumped into the straight tube 2 as needed to adjust the cell culture environment.

[0076] The specific implementation process is as follows:

[0077] First, by opening the solenoid valve at the top of the straight tube 2, the cells to be cultured are pumped into the straight tube 2 at a low speed. After settling, the control system starts the dual-head motor. The output shaft of one of the dual-head motors drives the main bevel gear 5 to rotate through the coupling. The main bevel gear 5 meshes with the secondary bevel gear 6, so that the secondary bevel gear 6 drives the straight tube 2 and the spiral tube 3 to rotate synchronously. This causes the cells to circulate and rise, and the spiral tube 3 rotates to promote cell suspension culture and prevent sedimentation.

[0078] The other output shaft of the dual-head motor drives the first gear 13 to rotate through a coupling. The first gear 13 meshes with the second gear 14, causing the eccentric actuating rod 16 of the second gear 14 to perform circular motion. Since the actuating rod 16 is embedded in the sliding groove of the moving seat 17, the circular motion is converted into the vertical reciprocating motion of the moving seat 17, so that the first rod 8 (nutrient solution control) and the second rod 11 (gas control) fixed on both sides of the moving seat 17 rise and fall synchronously.

[0079] The control system determines the current metabolic environment of the cell based on its growth stage or metabolic needs (e.g., high nutrient flow is required during the expansion phase); and dynamically generates control commands according to the needs. When nutrient solution needs to be delivered, the control system controls the telescopic component 18 to contract, causing the first gear 13 to mesh with the second gear 14. The second gear 14 then drives the movable seat 17 to move back and forth via the lever 16. At this time, the electromagnet 19 between the first rod 8 and the first plate 9 is activated, connecting the first rod 8 and the first plate 9, causing the movable seat 17 to drive the first plate 9 to move back and forth, thereby pumping the nutrient solution into the straight tube 2.

[0080] When only gas needs to be delivered, the electromagnet 19 of the first rod 8 and the first plate 9 is closed, and the electromagnet 19 of the second rod 11 and the second plate 12 is opened. When the moving seat 17 moves, it will not drive the first plate 9 to move back and forth, and the nutrient solution delivery channel is closed. The connection between the second rod 11 and the second plate 12 allows the moving seat 17 to drive the second plate 12 to move back and forth, thereby pumping the gas required by the cells into the straight tube 2 to promote cell culture.

[0081] When there is no need to deliver nutrient solution or gas, the telescopic component 18 is extended by the control system. The telescopic component 18 drives the second gear 14 to disengage from the first gear 13, so that the second gear 14 will not drive the moving seat 17 to move, thus cutting off the power transmission channel.

[0082] This embodiment, through the coordinated design of its components, allows for flexible adjustment of nutrient solution composition, gas concentration, and flow rate at different culture stages. This enables independent control of nutrient solution and gas delivery, reducing the risk of imbalances in other environmental factors due to adjustments in a single parameter. This regulatory capability not only shortens the culture cycle but also reduces the toxic effects of accumulated metabolic waste on cells, ensuring CAR-NK cells grow in optimal conditions and improving product consistency and safety.

[0083] Example 2:

[0084] The difference from the above embodiments is that a gas sensor and a pH sensor are also fixedly bonded to the inner wall of the spiral tube 3. The control system is used to receive the gas information and pH information emitted by the gas sensor and pH sensor, and control the opening and closing of the telescopic component 18 and the electromagnet 19 based on the gas information and pH information respectively.

[0085] The specific implementation process is as follows: Key parameters inside the spiral tube 3 are monitored by gas and pH sensors. The control system analyzes the data stream in real time and links the actuator. The gas concentration adjustment telescopic component 18 is started and stopped. The electromagnet 19 is switched on and off according to the pH fluctuation to match the nutrient solution pulse rhythm, ensuring that the cell metabolic microenvironment is in the optimal state. This realizes the change of the culture environment from passive maintenance to active adaptation, providing high-precision physiological support for CAR-NK cell expansion.

[0086] Example 3:

[0087] As attached Figure 1 As shown, the difference from the above embodiment is that both the straight tube 2 and the spiral tube 3 are made of transparent material, and an image sensor is fixedly connected to the bracket 1 with screws; the control system is used to receive the image information emitted by the image sensor, and control the opening and closing of the telescopic component 18 and the electromagnet 19 based on the image information.

[0088] The specific implementation process is as follows: The image sensor is used to capture cell morphology and distribution information in real time. The control system uses AI vision algorithm to analyze key indicators such as cell aggregation degree and adhesion state. The telescopic component 18 is linked to adjust the gas supply (such as increasing the O2 flow when aggregation is abnormal). The opening and closing frequency of the electromagnet 19 is controlled synchronously to optimize the nutrient pulse rhythm (such as reducing the flow rate when apoptosis signs appear) to provide growth protection for CAR-NK cells.

[0089] Example 4:

[0090] As attached Figure 7 As shown, the control system differs from the above embodiments in that it includes a data acquisition module for collecting information, a data analysis module for analyzing information, an execution control module for controlling the operation of the device, and an abnormal calibration module for continuously monitoring cell growth.

[0091] The functions of each module are as follows:

[0092] The data acquisition module is used to collect environmental parameters inside the helical tube 3 through gas sensors (such as optical dissolved oxygen sensors or infrared CO2 sensors) and pH sensors; to collect cell morphological distribution information through image sensors (such as cameras); and to transmit the collected environmental parameters and morphological distribution information to the data analysis module. In this embodiment, the environmental parameters and raw image data are packaged into JSON format via RS485 bus and transmitted to the data analysis module with low latency (<100ms).

[0093] The data analysis module receives data transmitted from the data acquisition module, uses a deep learning model to analyze cell density, viability, and CAR expression rate, and integrates gas information, pH information, and cell morphology distribution information to generate environmental regulation instructions and nutrient supply strategies, outputting the analysis results. In this embodiment, a convolutional neural network (CNN) is used to process image data, outputting cell density (cells / mL), viability (%), and CAR expression rate; a long short-term memory (LSTM) network model is used to integrate gas information (dissolved oxygen or CO2) and pH time-series data to predict glucose consumption rate and lactic acid accumulation trends.

[0094] The execution control module controls the start and stop of the telescopic component 18 and the electromagnet 19 based on the analysis results of the data analysis module, injects specific mixed gas as needed and delivers a quantitative amount of nutrient solution; and controls the output of the dual output component 4 to adjust the rotation speed of the straight tube 2 and the spiral tube 3 (e.g., set to 20 rpm during the amplification period). In this embodiment, the execution control module adopts a PLC controller, which integrates a PID algorithm to dynamically adjust the speed of the dual-head motor (15-30 rpm ± 1%).

[0095] The anomaly calibration module is used to set thresholds for cell growth status based on environmental parameters and morphological distribution information. When the threshold is reached, it controls the opening and closing of the corresponding dual output component 4, electromagnet 19, and telescopic component 18, adjusting the gas injection frequency, nutrient solution supply amount, and rotation parameters to maintain optimal growth status. In this embodiment, a basic threshold library is set (such as boundary threshold ranges like pH < 7.0, O2 > 20%, or cell aggregation > 60%), and the thresholds are dynamically adjusted according to historical amplification curves (e.g., the aggregation threshold rises to 70% on day 5).

[0096] Example 5:

[0097] As attached Figure 8 As shown, unlike the above embodiments, this embodiment also provides an AI-based CAR-NK cell culture method, including the following steps:

[0098] S1. Cell seeding and initial culture: Inject the CAR gene-modified NK cell suspension into the spiral tube 3 through the interface at the bottom of the straight tube 2 to form a closed culture environment; start the dual output device 4 to drive the straight tube 2 and spiral tube 3 to rotate, and set the initial speed to prevent cell sedimentation.

[0099] S2. Real-time sensing of multi-source data: The gas information (such as dissolved oxygen or CO2 concentration) in the spiral tube 3 is monitored in real time by a gas sensor, the pH of the culture medium is collected by a pH sensor, the cell morphology distribution image is captured by an image sensor, and each monitoring threshold is set.

[0100] S3. Dynamic Decision-Making and Control: If the gas information exceeds the preset parameter threshold, the regulating component will inject mixed gas into the spiral tube 3; if the pH value exceeds the preset acid-base range, the delivery component will replenish the nutrient solution to adjust the pH value range; when the cell density growth rate is lower than the preset threshold, the rotation speed of the dual output component 4 will be increased and the nutrient solution replenishment frequency will be increased.

[0101] Specifically, for example, if the dissolved oxygen level is below 5 mg / L or the CO2 concentration is above 7%, the drive regulating component injects a mixed gas (containing 5% CO2 and 95% air) into the spiral tube 3; if the pH value exceeds the range of 7.0-7.6, the drive delivery component replenishes bicarbonate buffer or a dedicated pH adjusting solution to adjust the pH. When the cell density growth rate is below a preset threshold, the rotation speed of the dual output component 4 is increased by 2-5 rpm and the nutrient solution replenishment frequency is increased; when cell viability is <85%, a fortified culture medium containing IL-15 / IL-21 is injected.

[0102] S4. Closed-loop adaptive amplification: Continuously compare the actual cell activity with the preset growth curve. If the actual cell activity exceeds the preset growth curve, adjust the injection frequency of the mixed gas (calculated in real time based on the dissolved oxygen change rate), the single replenishment volume of nutrient solution (e.g., 0.1-0.5 mL / time, determined based on pH offset and cell metabolic rate), and the rotation period of the spiral tube 3 (e.g., switch the variable speed rotation of 0.5-5 rpm every 30 minutes to promote uniform nutrient distribution).

[0103] S5. Harvesting and Quality Control: When cell density reaches the standard (e.g., 1×10⁻⁶), harvesting and quality control are carried out. 7 When the cell count reaches (≥70%), the solenoid valve is opened to collect the cell suspension; and the cytotoxic activity of the cells is verified by flow cytometry to complete batch production. In this embodiment, the activity and other indicators of CAR-NK cells are detected by flow cytometry, or the gene expression of CAR-NK cells is detected by gene sequencing and other methods.

[0104] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. An AI-based CAR-NK cell culture device, comprising a scaffold (1), characterized in that, It also includes a control system for controlling the operation of the device. A straight pipe (2) installed at an incline is rotatably connected to the support (1). Both ends of the straight pipe (2) have interfaces that communicate with the outside. Solenoid valves are connected to the interfaces. The control system is used to control the opening and closing of the solenoid valves. A spiral pipe (3) is connected to the straight pipe (2). A drive assembly for driving the straight pipe (2) to rotate is provided on the support (1). The drive assembly includes a dual output component (4) fixedly connected to the bottom of the bracket (1), and the control system is used to control the dual output component (4) to rotate; the output shaft of the dual output component (4) is coaxially fixedly connected to a main bevel gear (5), the main bevel gear (5) meshes with a secondary bevel gear (6), and the secondary bevel gear (6) is coaxially fixedly connected to the straight tube (2); The support (1) is also provided with a delivery component for delivering nutrient solution to the spiral tube (3) and a regulating component for regulating the gas environment inside the spiral tube (3); The drive assembly is equipped with a transmission assembly for driving the conveying assembly and the regulating assembly; the transmission assembly is equipped with an opening and closing assembly for driving the conveying assembly and the regulating assembly independently; the control system is used to control the operation of the transmission assembly and the opening and closing assembly respectively to regulate the cell growth environment.

2. The AI-based CAR-NK cell culture device according to claim 1, characterized in that, The conveying assembly includes a first cylinder (7), a first rod (8), and a first plate (9). The first cylinder (7) is fixedly connected to the bottom of the bracket (1). The first plate (9) slides vertically with the inner wall of the first cylinder (7). The first rod (8) is located above the first plate (9). The opening and closing assembly is used to connect the first rod (8) and the first plate (9). The transmission assembly is used to drive the first rod (8) to reciprocate. The bottom of the first cylinder (7) is connected to an input pipe and an output pipe, and the connection between the input pipe and the output pipe is connected to a first one-way valve; the end of the input pipe away from the first cylinder (7) is connected to a storage tank for storing nutrient solution, and the end of the output pipe away from the first cylinder (7) is connected to the inside of the straight pipe (2).

3. The AI-based CAR-NK cell culture device according to claim 2, characterized in that, The adjustment assembly includes a second cylinder (10), a second rod (11), and a second plate (12). The second cylinder (10) is fixedly connected to the side of the bracket (1) away from the first cylinder (7). The second plate (12) slides vertically against the inner wall of the second cylinder (10). The second rod (11) is located above the second plate (12). The opening and closing assembly is used to connect the second rod (11) and the second plate (12). The transmission assembly is used to drive the second rod (11) to reciprocate. The bottom of the second cylinder (10) is connected to an inlet pipe and an outlet pipe. The connection between the inlet pipe and the outlet pipe is connected to a second one-way valve. The end of the inlet pipe away from the second cylinder (10) is connected to a gas storage tank for storing gas, and the end of the outlet pipe away from the second cylinder (10) is connected to the inside of the straight pipe (2).

4. The AI-based CAR-NK cell culture device according to claim 3, characterized in that, The transmission assembly includes a first gear (13) and a second gear (14). The first gear (13) is coaxially fixedly connected to the side of the double output component (4) away from the main bevel gear (5). A fixed frame (15) is fixedly connected to the bracket (1), and the first gear (13) is rotatably connected to the fixed frame (15). The second gear (14) meshes with the first gear (13), and an actuating rod (16) is eccentrically fixedly connected to the second gear (14). A movable seat (17) is vertically slidably fitted on the fixed frame (15), and the first rod (8) and the second rod (11) are fixedly connected to both sides of the movable seat (17). A sliding groove is opened on the movable seat (17) for the actuating rod (16) to move. The bracket (1) is also provided with a telescopic assembly for disengaging the first gear (13) and the second gear (14).

5. The AI-based CAR-NK cell culture device according to claim 4, characterized in that, The telescopic assembly includes a telescopic member (18) fixedly connected to the bracket (1), and a control system is used to control the telescopic member (18) to extend and retract. The output shaft of the telescopic member (18) is rotatably connected to the second gear (14).

6. The AI-based CAR-NK cell culture device according to claim 5, characterized in that, The opening and closing assembly includes several electromagnets (19), and the control system is used to control the opening and closing of the electromagnets (19). The electromagnets (19) are fixedly connected to the bottom ends of the first rod (8) and the second rod (11), respectively. The first plate (9) and the second plate (12) are both embedded with magnetic blocks (20) that are magnetically matched with the electromagnets (19).

7. The AI-based CAR-NK cell culture device according to claim 6, characterized in that, A gas sensor and a pH sensor are also fixedly connected to the inner wall of the spiral tube (3). The control system is used to receive gas information and pH information emitted by the gas sensor and pH sensor, and to control the opening and closing of the telescopic component (18) and the electromagnet (19) based on the gas information and pH information respectively.

8. The AI-based CAR-NK cell culture device according to claim 7, characterized in that, Both the straight tube (2) and the spiral tube (3) are made of transparent material. An image sensor is also fixedly connected to the bracket (1). The control system is used to receive the image information emitted by the image sensor and control the opening and closing of the telescopic component (18) and the electromagnet (19) based on the image information.

9. The AI-based CAR-NK cell culture device according to claim 8, characterized in that, The control system includes the following modules: The data acquisition module is used to collect environmental parameters inside the spiral tube (3) through gas sensors and pH sensors; to collect cell morphology distribution information through image sensors; and to transmit the collected environmental parameters and morphology distribution information to the data analysis module. The data analysis module receives data transmitted from the data acquisition module, uses a deep learning model to analyze cell density, activity, and CAR expression rate, and integrates gas information, pH information, and cell morphology distribution information to generate environmental regulation instructions and nutrient supply strategies, and outputs analysis results. The execution control module is used to control the start and stop of the telescopic component (18) and the electromagnet (19) based on the analysis results of the data analysis module, inject specific mixed gas and deliver nutrient solution as needed; and control the output size of the dual output component (4) to adjust the rotation speed of the straight tube (2) and the spiral tube (3); An abnormal calibration module is used to set a threshold for cell growth status based on environmental parameters and morphological distribution information. When the threshold is reached, the corresponding dual output device (4), electromagnet (19) and telescopic device (18) are controlled to open and close, and the gas injection frequency, nutrient solution supply amount and rotation parameters are adjusted to maintain the best growth status.

10. An AI-based CAR-NK cell culture method, performed according to the AI-based CAR-NK cell culture apparatus according to any one of claims 1-9, characterized in that, Includes the following steps: S1. Cell seeding and initial culture: Inject the CAR gene-modified NK cell suspension into the spiral tube (3) through the interface at the bottom of the straight tube (2); start the dual output device (4) to drive the straight tube (2) and the spiral tube (3) to rotate, and set the initial rotation speed; S2. Real-time sensing of multi-source data: real-time monitoring of gas information in the spiral tube (3) by gas sensor, collection of acidity and alkalinity of culture medium by pH sensor, capture of cell morphology distribution image by image sensor, and setting of monitoring thresholds for each. S3. Dynamic decision-making and control: If the gas information exceeds the preset parameter threshold, the mixed gas is injected into the spiral tube (3) using the adjustment component; if the pH value exceeds the preset acid-base range, the nutrient solution is replenished using the delivery component to adjust the pH value range; when the cell density growth rate is lower than the preset threshold, the rotation speed of the dual output component (4) is increased and the nutrient solution replenishment frequency is increased. S4. Closed-loop adaptive amplification: continuously compare the actual cell activity with the preset growth curve. If the actual cell activity exceeds the preset growth curve, adjust the injection frequency of the mixed gas, the amount of nutrient solution supplied at one time, and the rotation period of the spiral tube (3). S5. Harvesting and Quality Control: When the cell density reaches the standard, the solenoid valve is opened to collect the cell suspension; and the killing activity of the cells is verified by flow cytometry to complete batch production.

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