Totally enclosed cell culture device and culture method

Abstract

The application relates to the technical field of cell culture, and discloses a fully-closed cell culture device, which comprises a three-dimensional culture bin, the inside of the three-dimensional culture bin is provided with a main culture space, the three-dimensional culture bin and a closable bin door structure are combined, the bin door structure comprises a lifting sealing door arranged on the front side of the culture bin body and a switchable partition located in the inside of the culture bin body, and the switchable partition divides the culture space into a plurality of array-arranged culture areas; the device adopts a square three-dimensional culture bin structure, fully utilizes the space geometric characteristics, realizes the maximized utilization of the internal culture space under the same land area, the square structure facilitates the array-arranged culture areas in the horizontal and vertical directions, improves the integration and loading density of the equipment, and significantly enhances the culture flux, so that an efficient and intensive physical platform is provided for multi-batch and large-scale cell culture.

Description

Technical Field

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

[0002] Cell therapy, as an important direction in modern biomedicine, has shown great potential in areas such as oncology, autoimmune diseases, and tissue repair. During in vitro culture, all types of cells generally rely on specific physicochemical environments to maintain their basic physiological functions. Among them, functional mammalian cells—including immune cells (such as T cells and NK cells), stem cells (such as mesenchymal stem cells and induced pluripotent stem cells), and genetically engineered cell therapy products (such as CAR-T and CAR-NK)—have more stringent requirements for in vitro culture conditions due to their highly specialized biological functions. These cells not only require stable temperature, humidity, and... Concentration is used to maintain pH and metabolic balance, and it is also extremely sensitive to microbial contamination, airflow disturbance and environmental fluctuations. Any slight deviation may affect its proliferation capacity, phenotypic stability and even therapeutic efficacy.

[0003] However, currently widely used traditional cell culture devices have significant limitations in meeting the culture requirements of highly sensitive cells. Most devices employ a single-chamber structure with a loose internal spatial layout and lack effective regional isolation mechanisms, leading to easy cross-interference between different culture batches. More critically, their environmental control systems suffer from sluggish response, limited precision in temperature, humidity, and gas concentration regulation, and difficulty in maintaining high stability over extended periods. During operation, loading, changing, or observing culture dishes typically requires manually opening the chamber door, causing direct contact between the internal environment and the external laboratory air, resulting in sudden temperature drops, humidity loss, and other adverse effects. Risks of leakage and potential contamination. Such disturbances may have limited impact on conventional adherent cells, but for functional mammalian cells, they are often sufficient to cause cellular stress, functional decline, or even culture failure.

[0004] Furthermore, traditional culture devices generally have low space utilization rates, with a limited number of effective culture sites per unit area, making it difficult to support high-throughput, multi-batch parallel culture needs. To meet the scale requirements of clinical-grade cell preparation, multiple devices are often required, not only occupying a large amount of cleanroom space but also requiring more manpower for operation and monitoring, significantly increasing production costs. Simultaneously, the lack of automated transport and environmental pre-adaptation mechanisms makes the entire culture process highly dependent on manual intervention, further increasing the probability of operational errors and contamination. Against the backdrop of accelerated standardization and industrialization of cell therapy products, the shortcomings of existing culture equipment in terms of environmental stability, operational secrecy, throughput efficiency, and economics have become key bottlenecks restricting the high-quality, low-cost, and large-scale preparation of functional mammalian cells. Summary of the Invention

[0005] The purpose of this invention is to provide a fully enclosed cell culture device to solve the problems mentioned in the background art.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a fully enclosed cell culture device, comprising:

[0007] A three-dimensional culture chamber has a main culture space inside, which consists of a culture chamber body and an openable and closable door structure. The door structure includes a liftable and sealed door located on the front side of the culture chamber body and an openable and closable partition located inside the culture chamber body. The openable and closable partition divides the main culture space into multiple culture areas arranged in an array.

[0008] The three-dimensional culture unit consists of multiple three-dimensional culture positions that can be raised and lowered within each of the culture areas;

[0009] A docking unit is correspondingly set below each of the culture zones and together with the lifting and sealing door, forms a sealed pre-culture space. The docking unit includes a docking chamber and doors on both sides thereon, one of which is an internal lifting door opposite to the lifting and sealing door.

[0010] A transport and positioning unit is provided for each culture zone and is used to receive and transport the culture dish from the docking unit to the corresponding three-dimensional culture position. The transport and positioning unit includes a lifting component and a receiving support fixed on the lifting component. The receiving support can extend into the docking chamber.

[0011] According to the above technical solution, it also includes:

[0012] The sensing control unit includes:

[0013] A temperature control plate and a humidification port are located at the bottom of the culture zone. Culture inlet;

[0014] The culture vent is located at the top of the culture zone;

[0015] Temperature and humidity sensors for the culture zone are installed within the culture zone.

[0016] Temperature control panel for the docking zone, humidification port for the docking zone, and docking zone are installed inside the docking chamber. Inlet, outlet of the docking area, temperature sensor of the docking area, and humidity sensor of the docking area.

[0017] According to the above technical solution, the switchable partition includes:

[0018] An outer partition plate is fixedly installed inside the culture chamber, and a first vent hole is opened on its side wall;

[0019] An inner partition plate is fitted inside the outer partition plate. The inner partition plate is connected to an electric telescopic rod located inside the outer partition plate, and a second vent hole corresponding to the first vent hole is opened on the side wall.

[0020] According to the above technical solution, the three-dimensional culture site includes:

[0021] A mirror-symmetrical arc-shaped snap-fit ​​seat is fixedly installed on a guide seat on the side wall of the culture zone, and the top of the arc-shaped snap-fit ​​seat is provided with a snap-fit ​​groove;

[0022] The servo motor has a guide rod fixedly mounted on its output end, and the guide rod is threadedly connected to the guide seat through a threaded transmission rod.

[0023] According to the above technical solution, the lifting assembly includes:

[0024] A lifting chamber fixed to the outside of the three-dimensional culture chamber;

[0025] The drive motor is fixed inside the lifting chamber, and its output end is connected to the lifting transmission screw;

[0026] The lifting seat is threadedly fitted with the lifting drive screw.

[0027] According to the above technical solution, the supporting component includes:

[0028] A hollow support frame fixed to the lifting base;

[0029] Telescopic drive component, fixed inside the hollow support frame;

[0030] A fixed plate is fixedly connected to the telescopic end of the telescopic drive component;

[0031] Connecting plates are symmetrically fixed to one side of the fixing plate;

[0032] A petri dish receiving plate is fixed to the end of the connecting plate, and a receiving groove is provided at its upper end;

[0033] The arc-shaped connector has an opening area in the middle for the petri dish receiving plate to move up and down.

[0034] According to the above technical solution, the bottom of the lifting sealing door is symmetrically provided with openings and sealing structures adapted to the connecting plate, and the sealing structure includes:

[0035] An elastic sealing plate is movable inside the opening, and its top is connected to the top wall of the opening by a compression spring.

[0036] The sealing gasket is fixed to the bottom of the elastic sealing plate.

[0037] According to the above technical solution, the culture dish receiving plate is provided with a first positioning component, including at least two sets of first positioning driving components that can move centripetally and a first pressure detection component disposed at its end, and a first magnetic element is embedded at the upper end of the culture dish receiving plate.

[0038] The arc-shaped card holder is provided with a second positioning component, including at least two sets of second positioning drive members that can move centripetally and a second pressure detection member located at its end, and a magnetic element is embedded at the upper end of the arc-shaped card holder.

[0039] According to the above technical solution, the top of the docking chamber is equipped with a rapid sterilization component, which includes:

[0040] The ultraviolet light source is fixed to the top of the docking chamber.

[0041] The atomizing nozzle is connected to an external disinfectant supply line.

[0042] A cell culture method, comprising the following steps:

[0043] S1, External Loading and Sealing

[0044] Open the external lifting and sealing door, and the operator can place the culture dish containing cells directly onto the receiving support in the pre-culture space through the external opening. Close the external lifting and sealing door to ensure that the pre-culture space forms an independent and closed environment, completely isolated from the main culture space and the outside.

[0045] S2, Sterilization and Pre-culture

[0046] Rapid sterilization is initiated by sterilizing the enclosed pre-culture space using a rapid sterilization component. Ultraviolet light irradiation sterilizes the space, while atomizing nozzles spray disinfectant. Simultaneous environmental control utilizes a sensor control unit to regulate pre-culture space environmental parameters. The docking zone temperature control board heats or cools the space, with real-time temperature feedback from a temperature sensor. Humidity is adjusted via the docking zone humidification port, monitored by a humidity sensor. Gas is injected into the inlet to the target concentration, and environmental parameters are quickly adjusted to be completely consistent with the main culture space within the micro-enclosed space.

[0047] S3, Transfer the culture dish to the main culture zone.

[0048] After confirming that the pre-culture environment is consistent with the main culture space, the internal lifting door between the docking unit and the main culture chamber is opened, and the transport positioning unit drives the receiving support to rise vertically, transporting the petri dish from the pre-culture space to the designated three-dimensional culture position in the target culture area.

[0049] S4, Independent control of petri dish fixation and environment.

[0050] A servo motor drives the arc-shaped locking seat to close, locking the culture dish through the locking groove. The supporting component descends and resets, closing the internal lifting door. The adjustable partitions can be switched on and off as needed. In independent culture mode, the partitions can be switched off to close, allowing independent control of temperature and humidity in each culture zone. In co-culture mode, the adjacent switchable partitions are activated to achieve environmental homogenization across multiple zones. The temperature control board in the culture zone is linked with the sensor to maintain constant temperature and humidity. Dynamically balance gas concentration between the inlet and outlet;

[0051] S5, Incubation complete and aseptic removal

[0052] Open the internal lifting door, the transport positioning unit rises to the target three-dimensional culture position, the arc-shaped locking seat is released, the receiving support receives the culture dish, the receiving support carries the culture dish down to the pre-culture space, close the internal lifting door, open the external lifting sealing door, remove the culture dish from the receiving support, and close the external lifting sealing door.

[0053] Compared with the prior art, the beneficial effects achieved by the present invention are:

[0054] (1) This device adopts a square three-dimensional culture chamber structure, which makes full use of the spatial geometric characteristics and maximizes the use of the internal culture space under the same floor area. The square structure facilitates the array arrangement of multiple culture zones in the horizontal and vertical directions, improves the integration and loading density of the equipment, and significantly enhances the culture throughput. Each culture zone is equipped with an independent docking unit and a transport positioning unit to form a modular operation architecture. This design supports multi-channel parallel operation. Different culture zones can independently complete the loading, sterilization, pre-adaptation and transfer operations of culture dishes without interference, effectively improving the overall culture throughput. At the same time, this structure is conducive to the internal airflow organization and uniform distribution of environmental parameters, reducing dead zones and providing an efficient and intensive physical platform for multi-batch, large-scale cell culture, reducing cell culture costs.

[0055] (2) The main culture space consists of multiple independent culture zones and is in a fully enclosed operation state. It is completely isolated from the external environment through switchable partitions and double-layer door structure. During normal culture, both the lifting and sealing door and the internal lifting door are kept closed to ensure that the main culture space is not directly connected to the outside air, avoiding interference from temperature fluctuations, humidity changes and microbial contamination on cell growth. This zero-contact design effectively maintains constant temperature, constant humidity and stability. The high-precision environment of the concentration meets the high sensitivity and strict requirements of cells to culture conditions.

[0056] (3) This device is designed with a small, enclosed pre-culture space that can only accommodate a single culture dish, which is conducive to achieving rapid and efficient sterilization and environmental control. By combining the rapid sterilization component with ultraviolet irradiation and atomization disinfection, the culture dish and local space can be completely sterilized in a short time. At the same time, the sensor control unit adjusts the temperature control plate of the docking area, the humidification port of the docking area, and the docking area in real time. The injection port allows the pre-culture space to quickly reach the same environmental parameters as the target culture area, completing the cell pre-adaptation process, shortening the preparation cycle, and improving operational efficiency.

[0057] (4) By limiting the pre-culture operation to an independent pre-culture space, the main culture space remains stable throughout the entire culture cycle and is not affected by front-end operations such as loading, sterilization or transfer of culture dishes. All externally introduced culture dishes must first be sterilized and pre-adapted to the environment in the pre-culture space before being transferred to the main culture area through the internal lifting door, forming a one-way clean logistics path. This layered control mechanism further strengthens the independence and cleanliness level of the main culture environment, providing a highly controllable safety guarantee for the long-term stable expansion of cells.

[0058] (5) This device divides the main culture space into multiple arrayed culture zones and combines them with on / off control that allows for flexible switching between independent and combined culture. Temperature, humidity, and humidity can be independently set in each culture zone. Environmental parameters such as concentration can meet the differentiated culture needs of different experimental batches or cell states. At the same time, when high-throughput and consistent culture is required, multiple culture zones can be connected by opening adjacent switchable partitions to share a unified environment, ensuring a high degree of uniformity in cell growth conditions and significantly improving the adaptability and efficiency of the equipment.

[0059] (6) This device achieves automated transfer and precise handover of culture dishes through the coordinated cooperation of the transport positioning unit and the three-dimensional culture position. The entire process is automated. The support component is vertically raised and lowered under the drive of the lifting component, and horizontally extended and retracted through the telescopic drive component to complete the precise positioning in three-dimensional space. The arc-shaped clamping seat is raised and lowered as a whole under the drive of the servo motor to smoothly support the culture dish and avoid clamping stress damage. The whole process does not require manual intervention, which improves the repeatability and safety of the operation. Attached Figure Description

[0060] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used in conjunction with embodiments of the invention to explain the invention and do not constitute a limitation thereof. In the drawings:

[0061] Figure 1 This is a first perspective view of the present invention;

[0062] Figure 2This is a second perspective view of the present invention;

[0063] Figure 3 This is a third perspective view of the present invention;

[0064] Figure 4 This is a first partial three-dimensional schematic diagram of the present invention;

[0065] Figure 5 This is a second partial perspective view of the present invention;

[0066] Figure 6 This is a third partial perspective view of the present invention;

[0067] Figure 7 This is a fourth partial perspective view of the present invention;

[0068] Figure 8 This is a fifth partial perspective view of the present invention;

[0069] Figure 9 This is a sixth partial perspective view of the present invention;

[0070] Figure 10 This is a third-dimensional schematic diagram of the seventh part of the present invention;

[0071] Figure 11 This is the eighth partial perspective view of the present invention;

[0072] Figure 12 This is a third-dimensional schematic diagram of the ninth part of the present invention;

[0073] Figure 13 This is the present invention. Figure 7 Enlarged view of point A in the middle;

[0074] Figure 14 This is the present invention. Figure 11 Enlarged view of point B in the middle;

[0075] Figure 15 This is the present invention. Figure 11 Enlarged view of point C in the middle;

[0076] In the diagram: 100-Three-dimensional culture chamber, 101-Cultivation chamber body, 102-Lifting sealing door, 102a-Opening, 102b-Elastic sealing plate, 102c-Compression spring, 102d-Sealing gasket, 103-Openable partition, 103a-Outer partition plate, 103b-First vent, 103c-Inner partition plate, 103d-Second vent, 103e-Electric telescopic rod, 104-Cultivation area, 200-Three-dimensional culture unit, 201-Three-dimensional culture position, 201a-Arc-shaped snap-fit ​​seat, 201a1-Snap-fit ​​groove, 201b-Guide seat, 201c-Servo motor, 201 d-Threaded drive rod, 300-Dating unit, 301-Dating chamber, 302-Internal lifting door, 400-Transportation positioning unit, 401-Lifting assembly, 401a-Lifting chamber, 401b-Drive motor, 401c-Lifting drive screw, 401d-Lifting seat, 402-Supporting support component, 402a-Hollowed support frame, 402b-Telescopic drive component, 402c-Fixing plate, 402d-Connecting plate, 402e-Cultural dish receiving plate, 402e1-Receiving groove, 500-Sensing control unit, 501-Cultural temperature control plate, 502-Cultural humidification port, 503-Cultural... Inlet port, 504 - Culture outlet, 505 - Culture zone temperature sensor, 506 - Culture zone humidity sensor, 507 - Docking zone temperature control board, 508 - Docking zone humidification port, 509 - Docking zone 510 - Air outlet of docking area, 511 - Temperature sensor of docking area, 512 - Humidity sensor of docking area, 600 - First positioning component, 601 - First positioning drive component, 602 - First pressure detection component, 603 - First magnetic element, 604 - Second positioning component, 605 - Second positioning drive component, 606 - Second pressure detection component, 607 - Second magnetic element, 700 - Rapid sterilization component, 701 - Ultraviolet light source, 702 - Atomizing nozzle. Detailed Implementation

[0077] 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.

[0078] Please see Figure 1-15 The present invention provides a technical solution: a fully enclosed cell culture device, comprising:

[0079] The three-dimensional culture chamber 100 has a main culture space inside, which is composed of a culture chamber body 101 and an openable and closable door structure. The door structure includes a lifting and sealing door 102 located on the front side of the culture chamber body 101 and an openable and closable partition 103 located inside the culture chamber body 101. The openable and closable partition 103 divides the main culture space into multiple arrayed culture areas 104.

[0080] The three-dimensional culture unit 200 is composed of a plurality of three-dimensional culture positions 201 that can be raised and lowered and arranged in each of the culture areas 104;

[0081] The docking unit 300 is correspondingly disposed below each of the culture zones 104 and together with the lifting sealing door 102, forms a sealed pre-culture space. The docking unit 300 includes a docking chamber 301 and a chamber door disposed on both sides thereon, wherein one chamber door is an internal lifting door 302 opposite to the lifting sealing door 102.

[0082] A transport positioning unit 400 is provided for each culture zone 104 and is used to receive and transport the culture dish from the docking unit 300 to the corresponding three-dimensional culture position 201. The transport positioning unit includes a lifting component 401 and a receiving support 402 fixed on the lifting component 401. The receiving support 402 can extend into the docking chamber 301.

[0083] Specifically, it also includes:

[0084] Sensing control unit 500, comprising:

[0085] The culture temperature control plate 501 and the culture humidification port 502 are located at the bottom of the culture zone 104. Note entry 503;

[0086] The culture vent 504 is located at the top of the culture zone 104;

[0087] A temperature sensor 505 and a humidity sensor 506 are installed in the culture zone 104.

[0088] Temperature control panel 507, humidification port 508, and docking area humidification port are installed inside docking chamber 301. Inlet 509, outlet 510 of docking area, temperature sensor 511 of docking area, and humidity sensor 512 of docking area;

[0089] The humidification port 502 is located at the bottom of the culture zone 104 and is used to release water vapor into the culture space to increase air humidity. This component is connected to the humidity control system and can precisely replenish water according to actual needs, preventing the cultured genes from evaporating and condensing, thereby ensuring the stability of the cell growth microenvironment. Inlet 503 is also located at the bottom of the culture zone 104, used to inject carbon dioxide gas into the culture space to maintain a suitable culture environment. The concentration is adjusted to stabilize the pH value of the culture medium and meet the acid-base balance requirements of cell metabolism. A culture vent 504 is set at the top of the culture zone 104. Its function is to discharge excess gas, metabolic waste gas or residual gas after sterilization accumulated during the culture process into the culture space, while maintaining the internal air pressure balance and preventing excessive positive pressure or negative pressure from affecting the sealing structure. The vent can be equipped with a filter to prevent external pollutants from flowing back in. To achieve accurate sensing of environmental parameters, a culture zone temperature sensor 505 and a culture zone humidity sensor 506 are also installed in the culture zone 104. The culture zone temperature sensor 505 is used to collect the temperature data inside the culture zone 104 in real time and feed the signal back to the control system as the basis for adjusting the working status of the culture temperature control plate 501. The culture zone humidity sensor 506 is used to monitor the relative humidity of the air to ensure the accuracy and timeliness of the humidification operation and avoid condensation caused by excessive humidity or dryness caused by excessively low humidity.

[0090] Inside the miniature docking chamber 301, a complete environmental control and sensing system is also configured to ensure the pre-acclimatization process of the sample before it enters the main culture area. The docking area temperature control plate 507 is located inside the docking chamber 301, and its function is to independently regulate the temperature inside the docking unit 300, ensuring it quickly matches the environment of the main culture area 104, thus avoiding cell stress caused by temperature differences. The docking area humidification port 508 is used to supply water vapor into the docking chamber 301, regulating local air humidity and ensuring that external air introduced during opening and closing does not cause sudden changes in humidity, thereby maintaining the continuity of the cell microenvironment. Inlet 509 is used to inject carbon dioxide into the docking chamber 301, gradually bringing the gas environment closer to the formal culture conditions and achieving a smooth transition of cells from the outside to the culture system. Outlet 510, located inside the docking chamber 301, is used to expel outside air, residual sterilization gases, or operational waste gases that may be introduced during docking, maintaining the cleanliness and stable airflow of the docking space. Temperature sensor 511 and humidity sensor 512 are used to monitor the temperature and humidity inside the docking chamber 301 in real time. These two sensors transmit the collected data to the control system for automatic adjustment of the docking temperature control plate 507, humidification port 508, and other components in the docking area. The working status of the 509 inlet ensures that the pre-culture space has reached the set environmental standards before sample transfer. Two independently controllable yet coordinated microenvironment control systems were constructed, which not only ensured the long-term stable operation of the main culture area, but also achieved rapid environmental matching and post-sterilization recovery of the docking area, effectively improving the continuity, safety and success rate of cell culture.

[0091] Specifically, the switchable partition 103 includes:

[0092] An outer partition plate 103a is fixedly installed inside the culture chamber 101, and a first ventilation hole 103b is opened on its side wall;

[0093] An inner partition plate 103c is installed inside the outer partition plate 103a. The inner partition plate 103c is connected to an electric telescopic rod 103e located inside the outer partition plate 103a, and a second vent 103d corresponding to the first vent 103b is provided on the side wall.

[0094] The switchable partition 103 is used to regulate the gas flow between different culture zones 104 inside the three-dimensional culture chamber 100, enabling independent control or coordinated communication of the environment in each culture zone to meet the differentiated environmental requirements of different batches and stages of cell culture. This structure consists of an outer partition plate 103a and an inner partition plate 103c, which work together to form an adjustable ventilation control mechanism. The outer partition plate 103a is fixedly installed inside the culture chamber 101, serving as the main support structure for the switchable partition 103. Its position is relatively static. A first ventilation hole 103b is provided on the side wall of the outer partition plate 103a. This hole is one of the main channels for gas flow between adjacent culture zones 104. When the first ventilation hole 103b connects with the inner partition plate 103c, the gas flow is controlled. When the second vent 103d on the partition plate 103c is aligned, gas can pass through, realizing airflow exchange between areas. When misaligned, gas flow is blocked, forming a physical isolation. The inner partition plate 103c is movably fitted inside the outer partition plate 103a and can move under the action of the electric telescopic rod 103e, thereby changing its relative position with the outer partition plate 103a. The inner partition plate 103c has a second vent 103d on its side wall, which corresponds structurally to the first vent 103b. By adjusting the position of the inner partition plate 103c, the degree of overlap between the second vent 103d and the first vent 103b can be controlled, thereby realizing continuous or graded adjustment of the ventilation area. The electric telescopic rod 103e is set on the outer partition plate 103a. Inside the partition plate 103a, which is connected to the inner partition plate 103c, the electric telescopic rod 103e drives the movement of the inner partition plate 103c. This component executes actions according to the control system commands, switching the inner partition plate 103c between three states: open, partially open, and closed. When multiple culture zones 104 require unified environmental parameters, the electric telescopic rod 103e moves the inner partition plate 103c to a position where the first vent 103b and the second vent 103d are completely aligned, forming a smooth airflow channel. When a certain culture zone 104 needs to operate independently, the electric telescopic rod 103e adjusts the inner partition plate 103c to a state where the two vents are misaligned, achieving airtight isolation of that area. This structural design allows the three-dimensional culture chamber 100 to simultaneously operate within the same device. Running multiple independent culture programs, such as experimental groups under different temperature gradients, CO2 concentrations, or humidity conditions, significantly improves the flexibility and efficiency of the equipment. At the same time, during sterilization or troubleshooting, the affected area can be limited by closing the switchable partition 103 to ensure the normal operation of other areas. The switchable partition 103 achieves precise control of the gas communication status between culture zones 104 through the nesting cooperation of the outer partition plate 103a and the inner partition plate 103c, combined with the relative displacement adjustment of the first vent 103b and the second vent 103d. With the automatic operation of the electric telescopic rod 103e, this structure supports multi-area environmental zoning management, enhancing the controllability, safety, and adaptability of the cell culture process.

[0095] Specifically, the stereotactic culture site 201 includes:

[0096] A mirror-symmetrical arc-shaped snap-fit ​​seat 201a is fixedly installed on a guide seat 201b on the side wall of the culture zone 104. The top of the arc-shaped snap-fit ​​seat 201a is provided with a snap-fit ​​groove 201a1.

[0097] The servo motor 201c has a guide rod fixedly installed at its output end. The guide rod is threadedly connected to the guide seat 201b through the threaded transmission rod 201d.

[0098] The three-dimensional culture station 201 is used to support and fix the culture dish. It has a lifting function to cooperate with the transport and positioning unit 400 to complete the automatic handover and positioning of the culture dish. This structure realizes vertical movement through the servo motor 201c, ensuring that the transport process is not affected by adjacent culture stations in the multi-layer three-dimensional culture environment, improving operational safety and docking accuracy. The three-dimensional culture station 201 includes a pair of mirror-symmetrical arc-shaped locking seats 201a. This component is fixedly installed on the guide seat 201b, and the guide seat 201b is connected to the culture area 1. The sidewall connection of 04 forms a stable support base. The inner surface of the arc-shaped locking seat 201a is arc-shaped, which fits the outer edge of the standard round culture dish and can provide good fit and support. A locking groove 201a1 is provided on the top of the arc-shaped locking seat 201a. This groove is used to support the edge of the culture dish, preventing it from falling off in the vertical direction, and to limit it in the horizontal direction, ensuring that the culture dish is placed stably and centered. When the servo motor 201c is started, its rotational motion is transmitted to the threaded transmission rod 201d through the guide rod. 201d and guide seat 201b form a helical pair. Their rotation is converted into linear motion along the axial direction, thereby driving the entire arc-shaped retaining seat 201a to vertically rise and fall along the path set by guide seat 201b. Under normal culture conditions, the target three-dimensional culture position 201 is in a lower position, ensuring unobstructed space above it and preventing interference with other culture positions or transport paths. When the transport positioning unit 400 carries the culture dish to above the target position, the control system activates the servo motor 201c, driving the threaded transmission rod 201d to move, causing... The arc-shaped locking seat 201a of this layer moves upward along the guide seat 201b and gradually approaches the culture dish held by the transport positioning unit 400. When the arc-shaped locking seat 201a rises to a position that contacts or is close to the bottom of the culture dish, the transport positioning unit 400 releases the support for the culture dish, and the locking groove 201a1 at the top of the arc-shaped locking seat 201a supports the weight of the culture dish, completing the handover. Subsequently, the servo motor 201c runs in reverse, driving the arc-shaped locking seat 201a and the culture dish to descend to the preset culture height and enter a stable culture state.

[0099] Specifically, the lifting assembly 401 includes:

[0100] A lifting chamber 401a fixed to the outside of the three-dimensional culture chamber 100;

[0101] The drive motor 401b is fixed inside the lifting chamber 401a, and its output end is connected to the lifting transmission screw 401c.

[0102] The lifting seat 401d is threadedly engaged with the lifting transmission screw 401c.

[0103] The lifting assembly 401 is used to achieve precise vertical movement of the transport positioning unit 400, enabling it to lift the culture dish from the docking unit 300 to the three-dimensional culture position 201 at different heights for docking and placement. The drive motor 401b is installed inside the lifting chamber 401a, serving as the power source for the lifting motion. Its output end is connected to the lifting transmission screw 401c, driving the lifting transmission screw 401c to rotate synchronously through rotation. The drive motor 401b has precise speed and position control capabilities, and can achieve forward and reverse rotation and multi-level speed adjustment according to set commands, thereby controlling the movement direction and speed of the lifting seat 401d to meet the positioning requirements at different heights. The lifting transmission screw 401c is arranged along the axial direction of the lifting chamber 401a, with one end connected to the output shaft of the drive motor 401b and the other end supported by a bearing, forming a stable rotation axis. When the drive motor 401b works, the lifting transmission screw 401c rotates accordingly, driven by the screw thread. The lifting seat 401d, which works in conjunction with the lifting seat 401d, moves linearly along the axial direction. The lifting seat 401d is connected to the lifting transmission screw 401c by a threaded connection. The lifting seat 401d has a threaded hole inside that matches the lifting transmission screw 401c. It is fixedly connected to the support member 402 on the outside. When the lifting transmission screw 401c rotates, the lifting seat 401d moves up and down along the guide structure of the lifting chamber 401a under the action of the threaded thrust. This movement drives the support member 402 and the culture dish receiving plate 402e on it to rise and fall synchronously, thereby realizing the precise vertical transport of the culture dish. The movement trajectory of the lifting seat 401d is limited by the guide mechanism inside the lifting chamber 401a to ensure that it remains stable and without deflection during the lifting process, avoiding the impact of shaking on the positioning accuracy of the culture dish. At the same time, this component supports multi-point positioning function and can accurately stop at the height position of the corresponding three-dimensional culture position 201. It works with the transport positioning unit 400 to complete the layer-by-layer distribution and retrieval of the culture dish.

[0104] Specifically, the receiving support 402 includes:

[0105] A hollow support frame 402a is fixed on the lifting base 401d;

[0106] The telescopic drive component 402b is fixed inside the hollow support frame 402a;

[0107] Fixed plate 402c is fixedly connected to the telescopic end of telescopic drive component 402b;

[0108] Connecting plate 402d is symmetrically fixed to one side of fixing plate 402c;

[0109] The petri dish receiving plate 402e is fixed to the end of the connecting plate 402d, and its upper end is provided with a receiving groove 402e1;

[0110] The arc-shaped connector 201a has an opening area 201a2 in the middle for the petri dish receiving plate 402e to move up and down;

[0111] The receiving support 402 is used in the transport and positioning unit 400 to stably receive and horizontally telescopically transport the culture dish. Its structural design takes into account lightweight, precise transmission, and space avoidance, ensuring that the culture dish can be successfully grasped, lifted, positioned, and released in a multi-layer three-dimensional culture environment. The hollow support frame 402a is fixed on the lifting seat 401d and serves as the basic load-bearing structure for the receiving support 402. This support frame adopts a hollow design, which reduces the overall weight while ensuring structural strength and reducing the load on the lifting assembly 401. It also facilitates the installation of the telescopic drive component 402b, which is installed inside the hollow support frame 402a, serving as the horizontal support. The power actuator drives the subsequent connecting components to complete the telescopic movement, enabling the lateral transfer of the culture dish between the docking unit 300 and the three-dimensional culture position 201. The telescopic drive component 402b has controllable stroke and thrust output, which can precisely control the telescopic speed and position to ensure smooth and reliable operation. The fixing plate 402c is fixedly connected to the telescopic end of the telescopic drive component 402b and moves synchronously with the movement of the telescopic drive component 402b. This plate acts as an intermediate connecting component, transmitting the telescopic driving force to the subsequent structure and maintaining a rigid connection during the movement to prevent displacement or jamming due to uneven force. The connecting plate 402d is symmetrically fixed to one side of the fixing plate 402c, forming a left-right pair. The arrangement forms a stable double-arm support structure, which improves the balance of the end load and avoids unilateral tilting when carrying the culture dish, ensuring that the culture dish remains horizontal during transportation. The culture dish receiving plate 402e is fixed to the end of the connecting plate 402d and is used to directly support the culture dish. Its upper end is provided with a receiving groove 402e1. The shape of the groove matches the bottom contour of the culture dish, which can limit the outer edge of the culture dish and prevent it from sliding or tipping over during transportation. The depth and width of the receiving groove 402e1 are moderate, which can not only firmly support the culture dish, but also facilitate smooth release during handover. In terms of structural fit, the arc-shaped locking seat 201a has an opening area in the middle. Domain 201a2 is a reserved vertical channel space for the culture dish receiving plate 402e to pass freely during the lifting and lowering process. When the transport positioning unit 400 delivers the culture dish to the target position, the culture dish receiving plate 402e can be raised from below into the arc-shaped locking seat 201a until the bottom of the culture dish contacts the locking groove 201a1 to complete the handover. After the handover is completed, the culture dish receiving plate 402e can be withdrawn to the side and detached through the opening area 201a2, providing space for subsequent operations. This not only avoids the movement interference between mechanical structures, but also ensures the smoothness and reliability of the handover process, providing an efficient and safe solution for material transfer in the automated cell culture process.

[0112] Specifically, the bottom of the lifting sealing door 102 is symmetrically provided with openings 102a adapted to the connecting plate 402d and a sealing structure, the sealing structure including:

[0113] An elastic sealing plate 102b is movable within the opening 102a, and its top is connected to the top wall of the opening 102a by a compression spring 102c.

[0114] The sealing gasket 102d is fixed to the bottom end of the elastic sealing plate 102b;

[0115] The lifting and sealing door 102, as an openable and closable component on the front side of the three-dimensional culture chamber 100, participates in forming the pre-culture space above the docking unit 300 during the culture process. This space is a critical transition area when the transport and positioning unit 400 transfers the culture dish from the docking unit 300 to the culture area 104. It needs to have good airtightness to maintain a constant temperature, humidity, and CO2 environment. To achieve this goal, the bottom of the lifting and sealing door 102 is provided with an opening 102a that cooperates with the transport and positioning unit 400 and a matching sealing structure to ensure reliable closure after the transport action is completed, thus ensuring the integrity of the pre-culture space. In the operation process, the transport and positioning unit 400 performs the action first. Its supporting member 402 is transported under the drive of the telescopic drive member 402b, and the culture dish is transferred to the culture area. The culture dish receiving plate 402e, along with the culture dish, is fed into the docking unit 300. At this time, the lifting sealing door 102 is still open and not fully closed, providing a passage for the transport positioning unit 400. After the culture dish receiving plate 402e rises to the predetermined position, the lifting sealing door 102 begins to descend until its bottom forms a closed fit with the top structure of the docking unit 300. As the lifting sealing door 102 falls, the opening 102a at its bottom aligns with the connecting plate 402d on the transport positioning unit 400, allowing the connecting plate 402d to pass through, so that the culture dish receiving plate 402e can remain stably in the pre-culture space. To ensure the sealing performance in this state, the bottom of the lifting sealing door 102 is provided with a sealing structure, which includes a movable structure... The opening 102a contains an elastic sealing plate 102b, a compression spring 102c, and a sealing gasket 102d. The elastic sealing plate 102b is located inside the opening 102a and can move vertically up and down. Its top is connected to the top wall of the opening 102a via the compression spring 102c, which provides a downward preload, keeping the elastic sealing plate 102b in a downward trend when there is no external interference. The sealing gasket 102d is fixed to the bottom end of the elastic sealing plate 102b and is made of flexible sealing material, possessing good fit and durability. When the lifting sealing door 102 completes its descent and together with the docking unit 300 to form a pre-cultivation space, the connecting plate 402d has passed through the opening 102a and is in a fixed position. At this time, the elastic sealing... Under the action of the compression spring 102c, plate 102b is pressed downward against the side wall or surrounding support structure of connecting plate 402d. Sealing gasket 102d tightly adheres to the contact surface, effectively sealing the gaps around opening 102a. This ensures that the space remains sealed even when transport components pass through. Connecting plate 402d, as a stationary penetrating component, maintains stable sealing pressure even with minor assembly errors or thermal expansion and contraction due to the compensation capability of compression spring 102c. During the entire culture dish transport process, transport positioning unit 400 is the first action unit. After the culture dish is transported and positioned, lifting sealing door 102 performs a lowering and closing action, ultimately working in conjunction with docking unit 300 to construct a complete micro pre-culture space.The internal environment of this space is independently controlled by a sensor control unit 500 to ensure temperature, humidity, and humidity levels are maintained. Once the concentration reaches the set value, it prepares the cell culture device for subsequent entry into the main culture zone 104. This design ensures the airtightness of the pre-culture space, enabling the transportation process and environmental control to operate in tandem, thereby improving the overall automation level and environmental stability of the cell culture device.

[0116] Specifically, the culture dish receiving plate 402e is provided with a first positioning component 600, including at least two sets of first positioning driving members 601 that can move centripetally and a first pressure detection member 602 provided at its end, and a first magnetic element 603 is embedded at the upper end of the culture dish receiving plate 402e.

[0117] The arc-shaped card holder 201a is provided with a second positioning component 604, including at least two sets of second positioning drive members 605 that can move centripetally and a second pressure detection member 606 provided at its end, and a second magnetic element 607 is embedded at the upper end of the arc-shaped card holder 201a.

[0118] The first positioning component 600 on the petri dish receiving plate 402e and the second positioning component 604 on the arc-shaped locking seat 201a together constitute a high-precision docking and positioning system. This system not only achieves precise positioning of the petri dish during transport but also features a structurally optimized design specifically for compatibility with co-cultured dishes, ensuring stable adaptation and reliable operation of different or composite culture containers within the automated system. The first positioning component 600, located on the petri dish receiving plate 402e, includes at least two sets of centripetally movable first positioning drive components 601. These first positioning drive components 601 are distributed circumferentially along the petri dish receiving plate 402e and can synchronously extend or retract towards the center under the drive of the control system to adapt to the edges of petri dishes with different outer diameters. Each first positioning drive component 601 has a first pressure detection component 602 at its end. It is used to sense the pressure generated when it comes into contact with the side wall of the culture dish in real time and feed the signal back to the control system to determine whether effective contact has been completed, whether there is off-center loading or tilting, and then adjust the stroke of each driving component to achieve centering positioning. The upper end of the culture dish receiving plate 402e is embedded with a first magnetic element 603. This element is a permanent magnet material or electromagnetic unit with stable magnetic field characteristics. Its main function is not simply to generate magnetic attraction with the upper component, but as part of the identification and adaptation mechanism, specifically serving the composite culture container with co-culture dish. Co-culture dish usually refers to a special container used for co-culture of multi-cell systems, such as double-layer chambers, insert culture plugs or other integrated microfluidic culture devices. These containers often have magnetic mating structures integrated at the bottom or side wall to facilitate rapid alignment and stable fixation in the automated system.

[0119] The second positioning component 604 is disposed on the arc-shaped mounting base 201a and includes at least two sets of centripetally movable second positioning drive components 605. These second positioning drive components 605 are symmetrically installed on both sides of the arc-shaped mounting base 201a and can move independently or synchronously in the radial direction. They are used to perform secondary positioning calibration of the culture dish from above. Each second positioning drive component 605 has a second pressure detection component 606 at its end, which is used to detect the pressure state when it contacts the outer edge of the culture dish and provide upper positioning feedback information. It forms a double-layer pressure monitoring network with the first pressure detection component 602 to improve the overall positioning accuracy. A second magnetic element 607 is embedded at the upper end of the arc-shaped mounting base 201a. This element corresponds in spatial position to the first magnetic element 603 on the culture dish receiving plate 402e. When using a compatible container with a co-culture dish, the container itself may not have a standard outer edge structure, or its center of gravity distribution may differ from that of a conventional culture dish. In this case, the traditional mechanical limiting method is difficult to ensure stable support. Through the magnetic coupling between the first magnetic element 603 and the second magnetic element 607, the container can be pre-guided into the correct position in a non-contact state, and auxiliary adsorption force can be provided during the handover process to prevent lightweight or suspended co-culture components from shifting or flipping. More importantly, this magnetic pairing design allows the system to identify different types of culture containers. For example, when the control system detects the expected magnetic response between the first magnetic element 603 and the second magnetic element 607, it can determine that the current object being operated on is a co-culture dish that supports magnetic positioning, thereby automatically calling the corresponding transport parameters, lifting speed, positioning stroke, and clamping. Strategy: If no magnetic response is detected, the standard culture dish procedure is followed. The setting of the first magnetic element 603 and the second magnetic element 607 is not only an auxiliary means of physical positioning, but also a key technical feature for realizing intelligent identification and adaptive compatibility of multiple types of culture containers. It enables this device to not only be compatible with conventional round culture dishes, but also to support various integrated, modular, and magnetically labeled co-culture systems. This significantly improves the versatility and flexibility of the device in complex cell experiment scenarios, and realizes accurate identification, guidance and stable handover of non-standard or composite culture containers, enhancing the adaptability of cell culture devices in diverse experimental applications.

[0120] Specifically, the top of the docking chamber 301 is provided with a rapid sterilization component 700, which includes:

[0121] The ultraviolet light source 701 is fixed to the top of the docking chamber 301;

[0122] Atomizing nozzle 702 is connected to an external disinfectant supply line;

[0123] The rapid sterilization component 700 installed on the top of the docking chamber 301 is used to perform efficient and comprehensive microbial sterilization treatment on the internal space of the docking unit 300 before the start of the culture operation. This ensures that the culture dishes carrying cells are sterile before entering the main culture environment, preventing external contamination from entering the three-dimensional culture chamber 100. This component adopts a dual sterilization method combining physical and chemical methods to improve sterilization effect and response speed. The ultraviolet light source 701 is fixed on the top of the docking chamber 301. The short-wave ultraviolet light emitted by it has a strong bactericidal ability and can destroy the DNA or RNA structure of microorganisms, causing them to... Having lost their ability to reproduce and become active, the light source covers the main interior space of the docking chamber 301, directly irradiating the surface, walls, and bottom of the petri dishes. During the sterilization stage, the ultraviolet light source 701 is activated and operates continuously for a certain period of time, effectively killing bacteria, viruses, and fungal spores attached to the outer surface of the petri dishes and in the air, achieving surface sterilization in a static environment. The atomizing nozzle 702 is also located at the top of the docking chamber 301 and connected to the external disinfectant supply pipeline, which can evenly spray the disinfectant into the interior space of the docking chamber 301 in the form of fine droplets. The disinfectant used is usually hydrogen peroxide. Ethanol or other broad-spectrum disinfectants proven suitable for cellular operating environments are used. The atomizing nozzle 702 atomizes the liquid into suspended particles, allowing the disinfectant components to fully diffuse into corners, crevices, and other areas difficult for direct ultraviolet light to reach, achieving full three-dimensional sterilization coverage. In actual operation, the rapid sterilization component 700 typically executes the process step-by-step according to a preset program. First, the atomizing nozzle 702 sprays disinfectant mist, wetting the entire cavity and bringing it into contact with the disinfectant components. After a period of settling to ensure sufficient action, the ultraviolet light source 701 is activated to further decompose residual organic matter under light conditions and enhance... It has a strong inactivation effect on microorganisms. Some disinfectants, such as hydrogen peroxide, can also generate active oxygen under ultraviolet irradiation, which enhances the synergistic bactericidal ability. After sterilization, the system can be ventilated through the air outlet 510 in the docking area to remove residual mist and volatile substances, ensuring environmental safety when entering the next stage. The sterilization component is located at the top of the docking chamber 301, which is conducive to the downward radiation of ultraviolet light to form the maximum irradiation area. At the same time, it allows the atomized droplets to settle naturally, improving space utilization and sterilization uniformity. In addition, the top installation method facilitates maintenance and cleaning, and avoids affecting the placement and transportation of the culture dishes below.

[0124] A cell culture method, comprising the following steps:

[0125] S1, External Loading and Sealing

[0126] Open the external lifting and sealing door 102, and the operator can place the culture dish containing cells directly onto the receiving support 402 in the pre-culture space through the external opening. Close the external lifting and sealing door 102 to ensure that the pre-culture space forms an independent and closed environment, completely isolated from the main culture space and the outside.

[0127] S2, Sterilization and Pre-culture

[0128] Rapid sterilization is initiated by sterilizing the enclosed pre-culture space through the rapid sterilization component 700, irradiating the space with ultraviolet light source 701, spraying disinfectant through atomizing nozzle 702, and simultaneously controlling the environment by using the sensor control unit 500 to regulate the environmental parameters of the pre-culture space. The docking area temperature control plate 507 heats or cools the space, and the temperature sensor 511 provides real-time feedback. The humidification port 508 in the docking area adjusts the humidity, which is monitored by the humidity sensor 512. The CO2 injection port 509 in the docking area injects gas to the target concentration. The environmental parameters in the micro-enclosed space are quickly adjusted to be completely consistent with those of the main culture space.

[0129] S3, Transfer the culture dish to the main culture zone.

[0130] After confirming that the pre-culture environment is consistent with the main culture space, the internal lifting door 302 between the docking unit 300 and the main culture chamber 100 is opened, and the transport positioning unit 400 drives the receiving support 402 to rise vertically, transporting the culture dish from the pre-culture space to the designated three-dimensional culture position 201 of the target culture area 104.

[0131] S4, Independent control of petri dish fixation and environment.

[0132] Servo motor 201c drives arc-shaped locking seat 201a to close, locking the culture dish through locking groove 201a1, the support component 402 descends and resets, and the internal lifting door 302 closes. The switchable partition 103 can be adjusted as needed. In independent culture mode, the switchable partition 103 is closed, and the temperature, humidity and CO2 concentration of each culture zone 104 are independently controlled. In joint culture mode, adjacent switchable partitions 103 are opened to achieve environmental homogenization in multiple zones. The temperature control board 501 of the culture zone is linked with the sensor to maintain constant temperature and humidity. The CO2 injection port 503 and the gas outlet 504 of the culture zone dynamically balance the gas concentration.

[0133] S5, Incubation complete and aseptic removal

[0134] Open the internal lifting door 302, the transport positioning unit 400 rises to the target three-dimensional culture position 201, the arc-shaped locking seat 201a is released, the receiving support 402 receives the culture dish, the receiving support 402 carries the culture dish down to the pre-culture space, close the internal lifting door 302, open the external lifting sealing door 102, take the culture dish out from the receiving support 402, and close the external lifting sealing door 102.

[0135] Working principle: This device is a fully enclosed cell culture device, mainly designed for functional mammalian cells that need to be expanded in vitro in a strictly sterile, constant temperature and humidity and stable gas environment. It is especially suitable for immune cells that have high sensitivity or high cleanliness requirements for the culture environment. It focuses on three core aspects: sterility assurance, precise environmental control and automated operation, to ensure that immune cells are always in a stable, clean and controllable growth environment during the in vitro expansion process.

[0136] During the initial operation phase, the external lifting and sealing door is opened, and the operator places the culture container containing immune cells onto the receiving support within the pre-culture space. The door is then closed, creating a completely sealed cavity isolated from the outside environment. At this time, the main culture space remains closed, and each culture zone within it maintains the predetermined constant temperature, humidity, and temperature. Concentration is not affected by external operations.

[0137] Subsequently, the device initiates a rapid sterilization program, irradiating the interior of the chamber with an integrated ultraviolet light source at the top, while simultaneously spraying disinfectant mist from atomizing nozzles. This achieves combined sterilization of the outer surface of the culture container and the inner wall of the pre-culture space. After sterilization, the sensor control unit immediately intervenes, adjusting the temperature control plate and humidification port in the docking area. By injecting the inlet, the environment of the pre-culture space is quickly adjusted to be completely consistent with that of the main culture space according to the set parameters of the target culture area, so that the immune cells can complete environmental pre-adaptation before transfer and avoid stress or functional damage caused by sudden changes in physical and chemical conditions.

[0138] Once the environmental parameters stabilize, the internal lifting door opens, connecting the pre-culture space and the main culture space. The transport and positioning unit then activates, with its lifting components driving the support members to rise vertically, transporting the culture container to the corresponding height in the designated culture area. Subsequently, the telescopic drive pushes the culture dish receiving plate horizontally into place, precisely positioning it within the arc-shaped locking seat of the three-dimensional culture position. During this process, the upper and lower positioning components work together, using pressure detection feedback and magnetic guidance mechanisms to ensure the culture container is centered and transferred smoothly. After the transfer is completed, the arc-shaped locking seat locks in place under the drive of a servo motor, the transport and positioning unit resets, the internal lifting door closes, and the main culture space returns to a fully enclosed state.

[0139] Throughout the culture process, each culture zone can operate independently or be jointly controlled through switchable partitions to maintain differentiated culture conditions or a uniform, highly consistent environment, according to experimental needs. The sensor control unit continuously monitors and dynamically adjusts temperature, humidity, and gas concentration to ensure that immune cells expand under optimal conditions. After culture, the device reverses the transport process to aseptically remove the culture containers. No manual entry into the main culture space is required throughout the process, minimizing the risk of contamination. This achieves automation, standardization, and high reliability in immune cell culture, making it particularly suitable for large-scale in vitro expansion of immune cell types such as NK cells, T cells, CAR-T, and CAR-NK, which have stringent requirements for environmental cleanliness and stability.

[0140] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0141] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A fully enclosed cell culture device, characterized in that, include: The three-dimensional culture chamber (100) has a main culture space inside, which is composed of a culture chamber body (101) and an openable and closable door structure. The door structure includes a lifting and sealing door (102) located on the front side of the culture chamber body (101) and an openable partition (103) located inside the culture chamber body (101). The openable partition (103) divides the main culture space into multiple arrayed culture areas (104). The three-dimensional culture unit (200) is composed of a plurality of three-dimensional culture positions (201) that can be raised and lowered within each of the culture areas (104); The docking unit (300) is correspondingly disposed below each of the culture areas (104) and together with the lifting sealing door (102) forms a sealed pre-culture space. The docking unit (300) includes a docking chamber (301) and a chamber door disposed on both sides thereof, wherein one chamber door is an internal lifting door (302) opposite to the lifting sealing door (102). A transport positioning unit (400) is provided for each culture zone (104) to receive and transport the culture dish from the docking unit (300) to the corresponding three-dimensional culture position (201). The transport positioning unit includes a lifting component (401) and a receiving support component (402) fixed on the lifting component (401). The receiving support component (402) can extend into the docking chamber (301) in a telescopic manner. Sensing control unit (500), comprising: The culture temperature control plate (501), culture humidification port (502), and culture [other components] are located at the bottom of the culture zone (104). Note entry (503); The culture vent (504) is located at the top of the culture zone (104). A temperature sensor (505) and a humidity sensor (506) for the culture area are installed in the culture area (104). The docking zone temperature control panel (507), docking zone humidification port (508), and docking zone are installed inside the docking chamber (301). Inlet (509), outlet (510) of docking area, temperature sensor (511) of docking area and humidity sensor (512) of docking area; The docking chamber (301) is equipped with a rapid sterilization component (700) at its top, which includes: An ultraviolet light source (701) is fixed to the top of the docking chamber (301); Atomizing nozzle (702) is connected to an external disinfectant supply line.

2. The apparatus according to claim 1, characterized in that: The switchable partition (103) includes: An outer partition plate (103a) is fixedly installed inside the culture chamber (101), and a first ventilation hole (103b) is opened on its side wall. An inner partition plate (103c) is installed inside the outer partition plate (103a). The inner partition plate (103c) is connected to an electric telescopic rod (103e) located inside the outer partition plate (103a), and a second vent (103d) is provided on the side wall corresponding to the first vent (103b).

3. The apparatus according to claim 1, characterized in that: The stereotactic culture site (201) includes: A mirror-symmetrical arc-shaped snap-fit ​​seat (201a) is fixedly installed on a guide seat (201b) on the side wall of the culture area (104). The top of the arc-shaped snap-fit ​​seat (201a) is provided with a snap-fit ​​groove (201a1). The servo motor (201c) has a guide rod fixedly installed at its output end. The guide rod is threadedly connected to the guide seat (201b) through a threaded transmission rod (201d).

4. The apparatus according to claim 3, characterized in that: The lifting assembly (401) includes: A lifting chamber (401a) fixed to the outside of the three-dimensional culture chamber (100); The drive motor (401b) is fixed inside the lifting chamber (401a), and its output end is connected to the lifting transmission screw (401c). The lifting seat (401d) is threadedly engaged with the lifting transmission screw (401c).

5. The apparatus according to claim 4, characterized in that: The receiving support (402) includes: A hollow support frame (402a) is fixed on the lifting base (401d); The telescopic drive component (402b) is fixed inside the hollow support frame (402a); The fixed plate (402c) is fixedly connected to the telescopic end of the telescopic drive component (402b); The connecting plate (402d) is symmetrically fixed to one side of the fixing plate (402c); The petri dish receiving plate (402e) is fixed to the end of the connecting plate (402d), and its upper end is provided with a receiving groove (402e1). The arc-shaped connector (201a) has an opening area (201a2) in the middle for the petri dish receiving plate (402e) to move up and down.

6. The apparatus according to claim 5, characterized in that: The bottom of the lifting sealing door (102) is symmetrically provided with openings (102a) and sealing structures adapted to the connecting plate (402d), the sealing structure including: An elastic sealing plate (102b) is movable inside the opening (102a), and its top is connected to the top wall of the opening (102a) by a compression spring (102c); The sealing gasket (102d) is fixed to the bottom end of the elastic sealing plate (102b).

7. The apparatus according to claim 5, characterized in that: The petri dish receiving plate (402e) is provided with a first positioning component (600), including at least two sets of first positioning driving members (601) that can move centripetally and a first pressure detection member (602) provided at its end, and a first magnetic element (603) is embedded at the upper end of the petri dish receiving plate (402e). The arc-shaped card holder (201a) is provided with a second positioning component (604), including at least two sets of second positioning drive members (605) that can move centripetally and a second pressure detection member (606) located at its end, and a second magnetic element (607) is embedded at the upper end of the arc-shaped card holder (201a).

8. The cell culture method according to any one of claims 1-7, characterized in that, Includes the following steps: S1, External Loading and Sealing Open the external lifting and sealing door (102), place the culture dish containing cells on the receiving support (402) in the pre-culture space, and close the external lifting and sealing door (102) to form a closed pre-culture space; S2, Sterilization and Pre-culture The pre-culture space is sterilized by a rapid sterilization assembly (700), and the environmental parameters of the pre-culture space are adjusted to be consistent with those of the main culture space by a sensor control unit (500). S3, Transfer the culture dish to the main culture area. Open the internal lifting door (302), and the transport positioning unit (400) drives the receiving support (402) to rise, transporting the culture dish to the target three-dimensional culture position (201). S4, Independent control of petri dish fixation and environment. The culture dish is fixed in the three-dimensional culture position (201), the support component (402) is reset, the internal lifting door (302) is closed, and the ventilation status of the culture area (104) is adjusted as needed through the switchable partition (103); S5, Culture complete and aseptic removal Open the internal lifting door (302), raise the support component (402) to the target three-dimensional culture position (201), release the culture dish, and lower the support component (402) carrying the culture dish to the pre-culture space. Close the internal lifting door (302) and open the external lifting sealing door (102) to take out the culture dish.

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