A suspended cell culture apparatus and a suspended cell culture method

Abstract

This invention relates to the field of cell culture technology, specifically to a suspension cell culture device and method. The device includes a constant-temperature culture chamber, a swing-type mounting mechanism, a sterile ventilation component, an environmental monitoring module, and a ventilation structure. The inner wall of the constant-temperature culture chamber is equipped with a heat-insulating layer. The swing-type mounting mechanism uses a motor to drive a bracket to perform low-amplitude, low-frequency reciprocating swings. The bracket is equipped with a limiting component to quickly fix multiple culture bottles. Each culture bottle is equipped with an independent sterile ventilation component at its opening, enabling control of the ventilation flux and sterile protection for each bottle. An environmental monitoring module and a sterilization ventilation structure are installed inside the chamber to maintain a uniform and clean environment. This invention, along with a standardized culture method, solves the problems of easy cell sedimentation, susceptibility to shear damage, uneven environment, susceptibility to bacterial contamination, and uncontrollable culture in traditional culture methods. It exhibits good batch stability, strong versatility, and is suitable for small-scale suspension cell culture in laboratories.

Description

Technical Field

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

[0002] Suspension cell culture is a core technical step in biopharmaceutical and cell biology research. Currently, small-scale suspension cell culture in laboratories mostly uses ordinary cell culture flasks for static culture or a general shaker for shaking culture. However, the existing culture methods still have the following shortcomings in practical applications: 1. In static culture mode, the culture medium in the bottle is in a static state, and the suspended cells are prone to rapid sedimentation and deposition, and a large number of them will gather and clump together at the bottom of the bottle, causing cell accumulation and hypoxia, a decrease in metabolic rate, and consequently, reduced cell activity and increased apoptosis, making it difficult to achieve efficient expansion. In contrast, the shaking drive of conventional shakers is simple, and the shaking angle and frequency cannot be precisely controlled. The turbulent state of the liquid in the bottle is unstable. It is easy to generate high liquid shear force due to excessive disturbance intensity, which can damage the fragile suspended cells. It is also easy to fail to completely avoid cell sedimentation due to insufficient disturbance.

[0003] Second, the ventilation structure of ordinary culture flasks is simple, making it difficult to control the amount of gas exchange evenly. The concentration of carbon dioxide and dissolved oxygen in the flasks are unevenly distributed, and the environment of the upper and lower culture media is significantly different, which ultimately leads to uneven cell growth. Summary of the Invention

[0004] The purpose of this invention is to provide a suspension cell culture device and a suspension cell culture method to solve the problems mentioned in the background art.

[0005] The objective of this invention can be achieved through the following technical solutions: A suspension cell culture device includes a constant temperature culture chamber, a chamber cover at the top of the chamber, a control panel on the outer side of the chamber cover, and a heat insulation layer on the inner wall of the constant temperature culture chamber; it also includes: The culture flask is equipped with a sterile, breathable component at its mouth; A swing-type mounting mechanism is fixedly installed on the bottom surface of the constant temperature incubation chamber, which is used to limit the installation of multiple culture bottles and to gently swing the multiple culture bottles. An environmental monitoring module is installed in the constant temperature incubation chamber; And the ventilation structure set on the constant temperature incubation chamber.

[0006] Furthermore, the swing-type installation mechanism includes two mounting seats fixedly installed on the bottom surface of the constant temperature incubation chamber, a bracket is provided between the two mounting seats, and a rotating shaft is fixedly installed on both sides of the bracket. The ends of the two rotating shafts away from the bracket are rotatably connected to the mounting seats at corresponding positions. A motor is fixedly mounted on the outside of one of the mounting bases, and a rotating shaft corresponding to the motor rotates through the mounting base and is fixedly connected to the output shaft end of the motor. The top surface of the bracket is evenly provided with multiple limiting grooves that are adapted to the culture flasks, and the top of the through groove is provided with a limiting component.

[0007] Furthermore, the limiting assembly includes a spring and a hinge seat; The spring is coaxially mounted on the top of the limiting groove, and an abutment ring is fixedly mounted on the top of the spring. Multiple hinge seats are provided, and the multiple hinge seats are fixedly installed on the top surface of the bracket, and the multiple hinge seats are circumferentially arrayed about the axis of the limiting groove.

[0008] Furthermore, a limiting ring is provided on the periphery of the culture bottle near the top. When the culture bottle is in the installation state, the culture bottle passes through the limiting groove, and the limiting ring and the abutment ring are pressed into contact. The spring is in a compressed state, and the bent parts of the multiple L-shaped locking rods are all locked on the top surface of the limiting ring. The restoring force of the spring makes the L-shaped locking rod and the limiting ring in a pressed contact state.

[0009] Furthermore, the sterile ventilation component includes a tube, and the inside of the tube is provided with a sealing limiting sleeve, an adjustable ventilation valve and a sterile ventilation membrane in sequence from bottom to top. The tube is sealed and snapped into the open end of the culture bottle through the sealing limiting sleeve.

[0010] Furthermore, the environmental monitoring module includes a temperature sensor, a humidity sensor, and a gas concentration monitoring probe inside the chamber, used to monitor environmental parameters in the constant temperature incubation chamber in real time.

[0011] Furthermore, the ventilation structure includes a through groove at one end of the constant temperature incubation chamber and a mounting cylinder that is fixedly installed through the other end of the constant temperature incubation chamber. A filter element one is provided in the through groove, a fan is installed in the mounting cylinder, and a filter element two is provided in the mounting cylinder. The filter element two is located on the side of the fan that is close to the inside of the constant temperature incubation chamber.

[0012] Another object of the present invention is to provide a method for culturing suspension cells, comprising the following steps: S1: Equipment disinfection and pretreatment: Wipe and disinfect the inside of the constant temperature incubation chamber, the racks and sterile ventilation components with ultraviolet sterilization, let it stand to purify the environment inside the chamber, and complete the sterile pretreatment. S2: Culture medium preparation and bottling: Select a suspension cell culture flask, clean and sterilize it, prepare the suspension cell culture medium under aseptic conditions and add it quantitatively into the culture flask; S3: Bottle installation and sealing: Secure the filled culture bottles one by one in the limiting groove of the bracket, align the sterile ventilation component at the bottle mouth, and close the cover to create a sealed culture space; S4: Culture environment pre-adjustment: Pre-set culture temperature, swing parameters, and ventilation parameters through the control panel, start the equipment to preheat and stabilize the pressure, so that the environment inside the chamber meets the cell inoculation standard; S5: Inoculation and dynamic culture: Under aseptic conditions, inoculate each culture flask with the suspended cell seed solution, seal the flask opening and start the swing mechanism to carry out constant temperature suspension culture by low-frequency small-amplitude reciprocating swing. During the culture process, monitor environmental parameters in real time and dynamically adjust them until the cells reach the target density. S6: Collection and Reset: After stopping the shaking and allowing the culture flask to stand still, remove the culture flask to complete the cell collection. Disinfect the equipment and store it for later use.

[0013] The beneficial effects of this invention are: 1. This invention adopts a low-amplitude, low-frequency, stable reciprocating oscillation driving method, which gently drives the culture medium in the bottle to flow slowly, forming a gentle liquid flow environment, solving the problems of suspended cells sinking, accumulating, and clumping during static culture; the liquid flow shear force is extremely low throughout the process, which will not damage the fragile suspended cells, effectively improving cell survival rate and proliferation efficiency.

[0014] 2. The closed constant temperature incubation chamber, combined with the uniformly driven rack structure, allows for batch control of the incubation environment of all culture flasks. The temperature, gas environment, and shaking state of each culture flask are completely consistent, resulting in small batch differences in cell culture and strong culture stability.

[0015] 3. This invention adopts a dual protection structure of overall sterilization and ventilation of the chamber and independent sterile ventilation of each bottle. The ventilation structure of the chamber achieves full filtration of incoming and outgoing air through double-sided filter elements to maintain a clean environment inside the chamber. Each culture bottle is independently equipped with a sterile ventilation component, which can control the ventilation flow and independently block the invasion of microorganisms, avoid cross-contamination between bottles, and meet the needs of long-term closed amplification culture.

[0016] 4. The spring-loaded limiting assembly allows for quick assembly and disassembly of culture bottles by pressing and locking, making operation simple and convenient. The circumferentially distributed L-shaped locking rods, combined with spring force, ensure a secure fixation, preventing the bottles from shifting, tipping, or sliding during shaking, making it suitable for rapid batch operations.

[0017] 5. It comes with a dedicated suspension cell culture process, with standardized and continuous steps, and aseptic operation throughout the process. The process is easy to replicate and facilitates reproducible experimental culture. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort. Figure 1 This is a schematic diagram of the overall structure of the present invention; Figure 2 This is a schematic diagram of the internal structure of the constant temperature incubation chamber in this invention; Figure 3 This is a three-dimensional schematic diagram of the swing-type installation mechanism in this invention; Figure 4 yes Figure 3 Enlarged view of section C; Figure 5 yes Figure 2 Enlarged view of section A; Figure 6 This is a schematic diagram of the culture flask structure in this invention; Figure 7 yes Figure 6 Enlarged view of section B; The attached figures are labeled as follows: 1-Constant temperature incubation chamber, 2-Control panel, 3-Mounting cylinder, 4-Bed cover, 5-Passage groove, 6-Filter element one, 7-Environmental monitoring module, 8-Filter element two, 10-Fan, 11-Mounting base, 12-Bracket, 13-Cultivation bottle, 14-Motor, 15-Limiting ring, 16-Passage pipe, 17-Adjustable venting valve, 18-Sterile venting membrane, 19-Sealing limiting sleeve, 20-Spring, 21-Abutting ring, 22-Hinge seat, 23-L-shaped locking rod, 24-Rotating shaft, 25-Limiting groove. Detailed Implementation

[0019] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] Example 1: Please see Figure 1 and Figure 2In this embodiment of the invention, a suspension cell culture device includes a constant temperature culture chamber 1, a chamber cover 4 at the top of the constant temperature culture chamber 1, a control panel 2 on the outer side of the chamber cover 4, and a heat insulation layer on the inner wall of the constant temperature culture chamber 1. The constant temperature heating and temperature control functions of the constant temperature culture chamber 1 are implemented using existing technologies such as conventional electric heating components and temperature control feedback loops. This invention does not improve the principle and structure of constant temperature regulation, so it will not be described in detail here. It also includes: Culture flask 13 has a sterile, breathable component installed at the sealed joint of its mouth. A swing-type installation mechanism is fixedly installed on the bottom surface of the constant temperature incubation chamber 1 to limit the installation of multiple culture bottles 13 and to gently swing the multiple culture bottles 13. An environmental monitoring module 7 is installed in the constant temperature incubation chamber 1; And the ventilation structure set on the constant temperature incubation chamber 1.

[0021] The constant temperature incubation chamber 1 is a closed chamber, which, together with the heat insulation layer on the inner wall, forms an independent constant temperature incubation space. It can isolate external dust, bacteria and temperature fluctuations, and provide a stable basic environment for cell culture. The swing-type mounting mechanism integrates the functions of fixing the culture flask and swinging drive, providing power for cell suspension; The aseptic and breathable components provide breathability control and aseptic protection for individual bottles. The environmental monitoring module 7 enables real-time sensing of the environment inside the warehouse; the ventilation structure enables the circulation and purification of air inside the warehouse.

[0022] This integrated setup combines temperature control, oscillation, ventilation, monitoring, and sterilization functions into one unit, solving the problems of fragmented functions and weak environmental control capabilities in existing culture equipment.

[0023] Example 2: Please see Figures 1-6 Based on Example 1, the swing-type installation mechanism includes two mounting seats 11 fixedly installed on the bottom surface of the constant temperature incubation chamber 1, a bracket 12 is provided between the two mounting seats 11, and a rotating shaft 24 is fixedly installed on both sides of the bracket 12. The ends of the two rotating shafts 24 away from the bracket 12 are rotatably connected to the mounting seats 11 at the corresponding positions. A motor 14 is fixedly mounted on the outside of a mounting base 11, and a rotating shaft 24 corresponding to the motor 14 rotates through the mounting base 11 and is fixedly connected to the output shaft end of the motor 14. The top surface of the bracket 12 is evenly provided with multiple limiting grooves 25 that are adapted to the culture bottle 13, and the top of the through groove 5 is provided with a limiting component.

[0024] The limiting assembly includes a spring 20 and a hinge seat 22; A spring 20 coaxially arranged is fixedly installed on the top of the limiting groove 25, and an abutment ring 21 is fixedly installed on the top of the spring 20; Multiple hinge seats 22 are provided, and multiple hinge seats 22 are fixedly installed on the top surface of the bracket 12, and multiple hinge seats 22 are circumferentially distributed about the axis of the limiting groove 25.

[0025] A limiting ring 15 is provided on the periphery of the culture bottle 13 near the top. When the culture bottle 13 is in the installation state, the culture bottle 13 passes through the limiting groove 25, and the limiting ring 15 is pressed into contact with the abutment ring 21. The spring 20 is in a compressed state, and the bent parts of multiple L-shaped locking rods 23 are all locked on the top surface of the limiting ring 15. The restoring elasticity of the spring 20 makes the L-shaped locking rods 23 and the limiting ring 15 in a pressed contact state.

[0026] During operation, the motor 14 outputs power, which drives the bracket 12 to make a low-amplitude, low-frequency, and stable reciprocating swing around the axis of the rotating shaft 24. All the culture bottles 13 on the bracket 12 swing synchronously, causing the culture medium inside the bottles to flow slowly, forming a gentle liquid flow environment.

[0027] When installing culture bottle 13, insert culture bottle 13 downward into limiting groove 25. Limiting ring 15 presses down on abutment ring 21 to compress spring 20. Rotate L-shaped locking rod 23 so that its bent part is aligned with the top surface of limiting ring 15. After releasing culture bottle 13, the restoring force of spring 20 pushes abutment ring 21 upward, so that limiting ring 15 is clamped between L-shaped locking rod 23 and abutment ring 21, completing quick fixation. When disassembling, press down on culture bottle 13 to release the lock. After turning out L-shaped locking rod 23, culture bottle 13 can be removed.

[0028] This structure solves the problems of cell sinking, accumulation, and clumping during static culture by small-amplitude, low-frequency reciprocating oscillation. The shear force of the liquid flow is extremely low throughout the process, which can effectively protect the suspended cells and improve cell survival rate and proliferation efficiency. On the other hand, the spring-type limiting component is easy to assemble and disassemble, and is firmly fixed. During the oscillation, the culture flask 13 will not shift, tip over, or slide. All culture flasks 13 move in the same way, ensuring the uniformity of batch culture.

[0029] Example 3: Please see Figure 6 and Figure 7 Based on Example 2, the aseptic ventilation component includes a tube 16. The tube 16 has a sealing limiting sleeve 19, an adjustable ventilation valve 17 and a aseptic ventilation membrane 18 arranged sequentially from bottom to top inside. The tube 16 is sealed and snapped to the open end of the culture bottle 13 through the sealing limiting sleeve 19.

[0030] The tube 16 forms a sealed connection with the mouth of the culture bottle 13 through the sealing and limiting sleeve 19. All gases entering and leaving the culture bottle 13 must flow along the internal passage of the tube 16: external gases first pass through the sterile breathable membrane 18 for sterilization and filtration, then the ventilation flow is regulated by the adjustable breathable valve 17, and finally enter the interior of the culture bottle 13 through the sealing and limiting sleeve 19; waste gases generated by cell metabolism are discharged in the opposite direction along the same path.

[0031] This structure can uniformly control the air permeability of each culture flask 13, precisely match the cell's respiratory and metabolic needs, and reasonably balance the oxygen and carbon dioxide content in the flask, avoiding differences in cell growth caused by uneven air permeability; at the same time, the sterile breathable membrane 18 can effectively block the invasion of external microorganisms such as bacteria, fungi, and mycoplasma, building an independent sterile protective barrier for each flask. Even if the environment inside the chamber is accidentally contaminated, it will not cause cross-contamination between flasks, which is suitable for the needs of long-term closed amplification culture of culture flasks.

[0032] Example 4: Based on Example 1, the environmental monitoring module 7 includes a temperature sensor, a humidity sensor, and a gas concentration monitoring probe inside the chamber, which are used to monitor the environmental parameters in the constant temperature incubation chamber 1 in real time.

[0033] The various probes in the environmental monitoring module 7 can collect parameters such as temperature, humidity, and carbon dioxide concentration in the constant temperature incubation chamber 1 in real time, and transmit the data to the control panel 2 for display. Operators can indirectly and accurately control the incubation environment status inside all culture bottles 13 through the monitoring data. This achieves visualized control of the incubation process, and parameters can be adjusted in a timely manner when abnormalities occur, reducing unnecessary manual intervention by opening the lids, lowering the risk of contamination by other microorganisms, and improving the reproducibility of batch culture, ensuring a stable and controllable incubation process.

[0034] Example 5: Please see Figure 1 and Figure 5 Figure 2 Based on Example 1, the ventilation structure includes a through groove 5 opened at one end of the constant temperature incubation chamber 1 and an installation cylinder 3 fixedly installed at the other end of the constant temperature incubation chamber 1. A filter element 6 is provided in the through groove 5, a fan 10 is installed in the installation cylinder 3, and a filter element 8 is provided in the installation cylinder 3. The filter element 8 is located on the side of the fan 10 close to the inside of the constant temperature incubation chamber 1.

[0035] When in operation, the fan 10 starts, and the outside air is filtered and sterilized by filter element 2 8 and then sent into the constant temperature incubation chamber 1. The exhaust gas in the chamber is filtered by filter element 1 6 in the channel 5 and then discharged, forming a directional air circulation channel to continuously refresh the air in the chamber and balance the gas distribution.

[0036] This structure can circulate, filter, and clean the air inside the constant temperature incubation chamber 1, balancing the gas atmosphere throughout the chamber and ensuring a uniform and balanced incubation environment for all culture bottles 13, further reducing batch culture differences. At the same time, filter element 6 and filter element 8 form a dual sterilization system for air intake and exhaust, which, together with the sterile ventilation components at the bottle openings of culture bottles 13, forms a dual sterile barrier for overall chamber protection and individual bottle protection, improving the sterility protection level of the equipment and effectively reducing the probability of contamination by other microorganisms.

[0037] Example 6: Please see Figures 1 to 7 This example provides a method for culturing suspension cells, including the following steps: S1. Equipment disinfection and pretreatment: Wipe and disinfect all parts of the constant temperature incubation chamber 1, bracket 12, and sterile and breathable components. Use ultraviolet irradiation to complete the overall sterilization. Let the chamber environment stand still to purify the environment. After completing the initial sterile pretreatment of the equipment, it is ready for use. S2. Preparation and bottling of culture medium: Select a suitable suspension cell culture flask 13, clean and sterilize the culture flask 13, prepare the suspension cell culture medium under sterile conditions, and quantitatively add it into the culture flask 13. S3. Bottle installation and chamber closure: The culture bottles 13 that have been filled are sequentially clamped and fixed in the limiting grooves 25 of the bracket 12. Aseptic ventilation components are installed at the bottle mouth of each culture bottle 13. After confirming that the installation is secure, the chamber cover 4 is closed to create a sealed and sterile culture space. S4. Pre-conditioning of the culture environment: By setting parameters such as culture temperature, shaking frequency, shaking amplitude, and air exchange cycle through the control panel 2, the equipment is started to preheat and stabilize the pressure in advance, so that the overall environment in the constant temperature culture chamber 1 reaches the standard conditions for suspension cell inoculation. S5. Inoculation and Dynamic Culture: Under aseptic conditions, inoculate each culture flask 13 with the suspended cell seed solution. After inoculation, seal the flask opening. Activate the swing-type installation mechanism to drive the culture medium to flow smoothly with a low-frequency, small-amplitude, gentle reciprocating shaking motion, ensuring uniform suspension and dispersion of cells, and entering the isothermal suspension culture stage. During the culture process, the equipment monitors the temperature, humidity, and gas concentration parameters in the chamber in real time, automatically fine-tuning the isothermal value, swing rhythm, and air exchange frequency to maintain a stable culture environment until the cells expand to the target density. S6. End of culture and equipment reset: After the cells reach the target density, stop the rocking drive and maintain a static buffer. Take out culture flask 13 in sequence to complete cell collection, passage or sampling. After the culture is completed, disinfect the inside of the equipment again and keep it for subsequent recycling.

[0038] This embodiment provides a standardized suspension cell culture method matched with the above-mentioned apparatus, and connects various functions of the apparatus in series through the procedure of "disinfection and sterilization - solution preparation - installation - pre-adjustment - inoculation - cultivation - collection". The method has standardized steps, achieves aseptic operation throughout the whole process, features strong reproducibility of the procedure, and facilitates the development of repeated experimental culture; when combined with the low-shear rocking, dual aseptic protection and real-time environmental control of the apparatus, the method is suitable for various scenarios such as scientific research experiments and small-scale cell amplification.

[0039] The foregoing shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and the description in the specification are only intended to illustrate the principle of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention, and all these changes and modifications fall within the scope of the claimed present invention.

Claims

1. A suspension cell culture device, comprising a constant temperature culture chamber (1), wherein a chamber cover (4) is provided at the top of the constant temperature culture chamber (1), a control panel (2) is provided on the outside of the chamber cover (4), and a heat insulation layer is provided on the inner wall of the constant temperature culture chamber (1); characterized in that, Also includes: Culture bottle (13), wherein a sterile air-permeable component is installed at the mouth of the culture bottle (13) in a sealed connection; A swing-type installation mechanism is fixedly installed on the bottom surface of the constant temperature incubation chamber (1) for limiting the installation of multiple culture bottles (13) and for gently swinging the multiple culture bottles (13); An environmental monitoring module (7) is installed in the constant temperature incubation chamber (1); And the ventilation structure set on the constant temperature incubation chamber (1).

2. The suspension cell culture device according to claim 1, characterized in that, The swing-type installation mechanism includes two mounting seats (11) fixedly installed on the bottom surface of the constant temperature incubation chamber (1), a bracket (12) is provided between the two mounting seats (11), and a rotating shaft (24) is fixedly installed on both sides of the bracket (12). The ends of the two rotating shafts (24) away from the bracket (12) are rotatably connected to the mounting seats (11) at the corresponding positions. A motor (14) is fixedly mounted on the outside of one of the mounting bases (11), and a rotating shaft (24) corresponding to the motor (14) rotates through the mounting base (11) and is fixedly connected to the output shaft end of the motor (14). The top surface of the bracket (12) is evenly provided with multiple limiting grooves (25) that are adapted to the culture bottle (13), and the top of the two through grooves (5) is provided with a limiting component.

3. The suspension cell culture device according to claim 2, characterized in that, The limiting component includes a spring (20) and a hinge seat (22); The spring (20) is fixedly installed on the top of the limiting groove (25) and the spring (20) is fixedly installed on the top of the spring (20). Multiple hinge seats (22) are provided, and multiple hinge seats (22) are fixedly installed on the top surface of the bracket (12), and multiple hinge seats (22) are circumferentially distributed about the axis of the limiting groove (25).

4. The suspension cell culture device according to claim 3, characterized in that, The culture bottle (13) has a limiting ring (15) near the top on its periphery. When the culture bottle (13) is in the installation state, the culture bottle (13) passes through the limiting groove (25), the limiting ring (15) and the abutment ring (21) are pressed together, the spring (20) is in a compressed state, and the bent parts of the multiple L-shaped locking rods (23) are all stuck on the top surface of the limiting ring (15). The restoring force of the spring (20) makes the L-shaped locking rods (23) and the limiting ring (15) in a pressed contact state.

5. The suspension cell culture device according to claim 1, characterized in that, The sterile ventilation assembly includes a tube (16), inside which a sealing limiting sleeve (19), an adjustable ventilation valve (17) and a sterile ventilation membrane (18) are arranged sequentially from bottom to top. The tube (16) is sealed and snapped to the open end of the culture bottle (13) through the sealing limiting sleeve (19).

6. The suspension cell culture device according to claim 1, characterized in that, The environmental monitoring module (7) includes a temperature sensor, a humidity sensor, and a gas concentration monitoring probe in the chamber, which is used to monitor the environmental parameters in the constant temperature incubation chamber (1) in real time.

7. The suspension cell culture device according to claim 1, characterized in that, The ventilation structure includes a through groove (5) at one end of the constant temperature incubation chamber (1) and an installation cylinder (3) fixedly installed at the other end of the constant temperature incubation chamber (1). A filter element (6) is provided in the through groove (5), a fan (10) is installed in the installation cylinder (3), and a filter element (8) is provided in the installation cylinder (3). The filter element (8) is located on the side of the fan (10) close to the inside of the constant temperature incubation chamber (1).

8. A method for culturing suspension cells, implemented using the suspension cell culture apparatus according to any one of claims 1-7, characterized in that, Includes the following steps S1: Equipment disinfection and pretreatment: Wipe and disinfect the inside of the constant temperature incubation chamber, the racks and sterile ventilation components with ultraviolet sterilization, let it stand to purify the environment inside the chamber, and complete the sterile pretreatment. S2: Culture medium preparation and bottling: Select a suspension cell culture flask, clean and sterilize it, prepare the suspension cell culture medium under aseptic conditions and add it quantitatively into the culture flask; S3: Bottle installation and sealing: Secure the filled culture bottles one by one in the limiting groove of the bracket, align the sterile ventilation component at the bottle mouth, and close the cover to create a sealed culture space; S4: Culture environment pre-adjustment: Pre-set the culture temperature, swing parameters, and ventilation parameters through the control panel, start the equipment to preheat and stabilize the pressure, so that the environment inside the chamber meets the cell inoculation standard; S5: Inoculation and dynamic culture: Under aseptic conditions, inoculate each culture flask with the suspended cell seed solution, seal the flask opening and start the swing mechanism to carry out constant temperature suspension culture by low-frequency small-amplitude reciprocating swing. During the culture process, monitor environmental parameters in real time and dynamically adjust them until the cells reach the target density. S6: Collection and Reset: After stopping the shaking and allowing the culture flask to stand still, remove the culture flask to complete the cell collection. Disinfect the equipment and store it for later use.

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