Circuitry for facilitating disassembly of a cell culture device

Abstract

The application discloses a convenient dismounting loop type cell culture device. The application sets the convenient dismounting loop type cell culture device to include a cell culture cavity, an intermediate layer partition plate and a liquid supply and sampling cavity, realizes a sealed environment of a cell culture process, avoids a pollution risk, sets an integrated monitoring device in the cell culture cavity to realize monitoring of a culture bottle placed above the integrated monitoring device, sets a liquid supplement peristaltic pump, an alkali supplement peristaltic pump and a sampling peristaltic pump in the liquid supply and sampling cavity, cooperates with a control host to realize real-time supplement of culture liquid or alkali liquid in the culture bottle based on monitoring, can extract cell liquid in the culture bottle for sampling at a suitable time, can completely realize automatic processing, and the liquid supplement loop, the alkali supplement loop, the sampling loop and the culture bottle are convenient to dismount and replace, improve a success rate and efficiency of cell culture, reduce manual intervention and reduce a pollution risk.

Description

Technical Field

[0001] This application relates to the field of cell culture technology, specifically to a conveniently detachable loop cell culture device. Background Technology

[0002] Traditional cell culture monitoring systems primarily focus on monitoring single parameters, such as systems that only monitor oxygen concentration. Taking a detachable loop cell culture device with only medium replenishment as an example, it typically consists of a medium storage unit, a peristaltic pump, and connecting tubing. The medium storage unit is usually a sealed container for holding the cell culture medium. The peristaltic pump, connected to the medium storage unit via tubing, generates pressure to propel the medium through the tubing into the cell culture flask. Valves may be installed on the tubing to control the volume and flow rate of the medium. To accurately control cell growth, operations such as replenishing the medium and alkali solution, and sample collection must be performed manually, increasing the risk of contamination and workload. Furthermore, after a period of use, the culture flask, medium storage unit, alkali solution bottle, and connecting tubing need to be replaced, which is difficult due to the inconvenience of disassembly. Therefore, there is currently a lack of easily disassembled loop cell culture devices that can comprehensively monitor cell culture process data and automatically perform operations such as replenishing culture medium and alkali, and sample collection. These devices should also allow for easy disassembly and replacement of culture flasks, culture medium storage devices, alkali bottles, and connecting tubing to improve the success rate and efficiency of cell culture, reduce human intervention, and lower the risk of contamination. Summary of the Invention

[0003] This application provides a convenient disassembly loop cell culture device, which can solve the technical problems of current convenient disassembly loop cell culture devices lacking the ability to comprehensively monitor cell culture process data and automatically perform operations such as replenishing culture medium and alkali, and sample collection. It is also inconvenient to disassemble and replace culture flasks, culture medium storage devices, alkali bottles and connecting pipelines, resulting in low cell culture success rate and efficiency, and increased risk of contamination and workload.

[0004] This application provides a conveniently disassembled loop cell culture device, including:

[0005] The cell culture chamber is equipped with a sealed door;

[0006] The shaking device is located inside the cell culture chamber;

[0007] An integrated monitoring device is disposed within the cell culture chamber and is mounted on the shaker device;

[0008] At least one culture flask is disposed within the cell culture chamber and located above the integrated monitoring device;

[0009] An intermediate partition is disposed at the top of the cell culture chamber; the intermediate partition is provided with a sealing communication hole;

[0010] A liquid supply and sampling chamber is located above the cell culture chamber; the liquid supply and sampling chamber is connected to the cell culture chamber through the sealed communication hole;

[0011] A replenishment circuit is provided in the liquid supply and sampling chamber, and the replenishment circuit is connected to the culture flask through the sealed communication hole;

[0012] An alkali replenishment circuit is provided in the liquid supply sampling chamber, and the alkali replenishment circuit is connected to the culture flask through the sealed communication hole;

[0013] A sampling circuit is provided in the liquid supply sampling chamber, and the sampling circuit is connected to the culture flask through the sealed communication hole;

[0014] The control host is located outside the liquid supply and sampling chamber, and the control host is connected to the integrated monitoring device, the liquid replenishment circuit, the alkali replenishment circuit and the sampling circuit.

[0015] Furthermore, the culture flask is equipped with a sealing cap at its opening, and the sealing cap has three connecting tubes, which are respectively connected to the liquid replenishment circuit, the alkali replenishment circuit, and the sampling circuit.

[0016] Furthermore, the fluid replenishment circuit includes:

[0017] At least one replenishment bottle is fixed inside the liquid supply and sampling chamber;

[0018] At least one rehydration peristaltic pump is provided in the liquid supply and sampling chamber. The rehydration peristaltic pump is used to draw culture medium from the rehydration bottle and replenish it into the culture bottle.

[0019] The alkali replenishment circuit includes:

[0020] At least one alkali replenishment bottle is fixed inside the liquid supply sampling chamber;

[0021] At least one alkali replenishment peristaltic pump is installed in the liquid supply sampling chamber. The alkali replenishment peristaltic pump is used to draw alkali solution from the alkali replenishment bottle and replenish it to the culture bottle.

[0022] The sampling circuit includes:

[0023] At least one sampling peristaltic pump is disposed in the liquid supply sampling chamber, and the sampling peristaltic pump is used to draw cell fluid from the culture flask;

[0024] A sampling and dispensing device is connected to the sampling peristaltic pump, and the sampling and dispensing device distributes the cell fluid extracted by the sampling peristaltic pump into the sampling tube.

[0025] Furthermore, the liquid supply sampling chamber is also provided with a culture medium bottle holder, an alkali bottle holder, and a liquid tube holder. The culture medium bottle holder is located adjacent to the replenishment peristaltic pump, and the replenishment bottle is fixed on the culture medium bottle holder. The alkali bottle holder is located adjacent to the alkali replenishment peristaltic pump, and the alkali replenishment bottle is fixed on the alkali bottle holder. The liquid tube holder is provided with a liquid tube fixing hole.

[0026] Furthermore, the easily detachable loop cell culture device also includes:

[0027] A cooling module is located within the liquid supply and sampling chamber and is positioned corresponding to the culture medium bottle fixing seat.

[0028] Furthermore, there are four culture bottles, which are evenly distributed on the integrated monitoring device; the alkali replenishment bottle and the liquid replenishment peristaltic pump are connected to the culture bottles one by one, and the sampling peristaltic pump is connected to the culture bottles one by one.

[0029] Furthermore, the sampling and dispensing device includes:

[0030] The liquid separation module is provided with an inlet and multiple outlets. The inlet is connected to the sampling peristaltic pump, and the outlets are located on the lower surface of the liquid separation module.

[0031] A sampling box is removable and installed in the liquid supply sampling chamber. The sampling box is provided with a sampling slot, which is arranged along the extension direction of the liquid dispensing module and is located below the liquid outlet.

[0032] A sampling tube is inserted into the sampling slot and receives cell fluid flowing out from the outlet.

[0033] Furthermore, the sampling and dispensing device also includes:

[0034] A waste liquid pool is located inside the sampling box, arranged along the extension direction of the liquid separation module, and parallel to the sampling slot;

[0035] A transverse drive structure is connected to the liquid dispensing module to control the liquid dispensing module to move along the bottom plate direction parallel to the liquid supply sampling chamber. The transverse drive structure drives the liquid dispensing module to move between the sampling slot and the waste liquid pool.

[0036] Furthermore, the top of the cell culture chamber is provided with a guide groove corresponding to the sealed communication hole, and the intermediate partition includes a liquid tube clamping device. The liquid tube clamping device is located on the bottom plate of the liquid supply and sampling chamber and is located on one side of the sealed communication hole. A first liquid tube contour groove is provided on the other side of the sealed communication hole, and a second liquid tube contour groove is provided on the liquid tube clamping device corresponding to the first liquid tube contour groove. The first liquid tube contour groove and the second liquid tube contour groove form the sealed communication hole, and the liquid tube clamping device presses and seals the liquid tube in the sealed communication hole by pushing and pulling.

[0037] Furthermore, the liquid tube clamping device includes a rotary knob, a rotating shaft, a cam, a push plate, and a reset elastic element; the rotating shaft is perpendicularly arranged to the bottom plate of the liquid supply sampling chamber, the cam is connected to the middle of the rotating shaft, the rotary knob is connected to the top of the rotating shaft, the push plate abuts against the cam, the reset elastic element connects the bottom plate of the liquid supply sampling chamber to the push plate, and the push plate has a second liquid tube conforming groove on the side away from the reset elastic element, clamping the liquid tube between the second liquid tube conforming groove and the first liquid tube conforming groove.

[0038] The easily detachable loop-type cell culture device provided in this application embodiment, by setting up an easily detachable loop-type cell culture device including a cell culture chamber, an intermediate partition, and a liquid supply and sampling chamber, achieves a sealed environment for the cell culture process, avoiding the risk of contamination. Moreover, the integrated monitoring device set in the cell culture chamber can monitor the culture flasks placed above the integrated monitoring device, and the liquid supply and sampling chamber is equipped with a liquid replenishment peristaltic pump, an alkali replenishment peristaltic pump, and a sampling peristaltic pump. In conjunction with the control host, it can realize the real-time replenishment of culture medium or alkali based on the monitored conditions in the culture flasks, and can extract cell fluid from the culture flasks for sampling at appropriate times. It can achieve fully automated processing, and the liquid replenishment circuit, alkali replenishment circuit, sampling circuit, and culture flasks are easy to disassemble and replace, improving the success rate and efficiency of cell culture, reducing manual intervention, and reducing the risk of contamination. Attached Figure Description

[0039] The technical solution and other beneficial effects of this application will become apparent from the following detailed description of specific embodiments in conjunction with the accompanying drawings.

[0040] Figure 1 A schematic diagram showing the connection between a storage bottle, a waste bottle, a peristaltic pump, and a culture bottle in the existing technology;

[0041] Figure 2 This is a schematic diagram of the structure of the easily disassembled loop cell culture device provided in the embodiments of this application;

[0042] Figure 3A schematic diagram of the internal structure of the easily disassembled loop-type cell culture device provided in the embodiments of this application;

[0043] Figure 4 A schematic diagram of the structure of the easily disassembled loop-type cell culture device provided in the embodiments of this application with the tubing structure removed;

[0044] Figure 5 This is a schematic diagram of the internal structure of the liquid supply and sampling chamber provided in the embodiments of this application;

[0045] Figure 6 This is a schematic diagram of the structure of the intermediate layer partition provided in the embodiments of this application;

[0046] Figure 7 This is a schematic diagram of the structure of the liquid pipe clamping device provided in the embodiments of this application;

[0047] Figure 8 This is a schematic diagram of the structure of the cell culture chamber provided in the embodiments of this application;

[0048] Figure 9 This is a schematic diagram of the structure of the sealing cap of the culture flask provided in an embodiment of this application.

[0049] The markings in the diagram are as follows:

[0050] Cell culture chamber 1, sealed door 111, guide groove 112, inner protective shell 113, intermediate layer partition 2, sealed connecting hole 21, first liquid tube contour groove 211, second liquid tube contour groove 212, liquid tube clamping device 22, rotary knob 221, rotating shaft 222, cam 223, push plate 224, reset elastic element 225, liquid supply and sampling chamber 3, culture medium bottle fixing seat 31, alkali bottle fixing seat 32, liquid tube fixing bracket 33, fixing bracket 34, integrated monitoring device 4, culture bottle 5, sealed bottle cap 51, connecting pipe 52, liquid replenishment peristaltic pump 6, alkali replenishment peristaltic pump 7, sampling peristaltic pump 8, control host 9, sampling and dispensing device 10, dispensing module 101, sampling box 102, sampling card slot 103, waste liquid pool 104, transverse movement drive structure 105, shaking device 11, liquid replenishment bottle 12, alkali replenishment bottle 13, refrigeration module 14. Detailed Implementation

[0051] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are within the scope of protection of this application.

[0052] In the description of this application, it should be understood that the terms "center," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," and "counterclockwise," etc., indicating orientation or positional relationships based on the orientation or positional relationships shown in the accompanying drawings, are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this application. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, features defined with "first" and "second" may explicitly or implicitly include one or more of the stated features. In the description of this application, "a plurality of" means two or more, unless otherwise explicitly specified.

[0053] In the description of this application, it should be noted that, unless otherwise expressly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection, an electrical connection, or a connection that allows communication between them; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication between two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances.

[0054] In this application, unless otherwise expressly specified and limited, "above" or "below" the second feature can include direct contact between the first and second features, or contact between the first and second features through another feature between them. Furthermore, "above," "over," and "on top" of the second feature includes the first feature being directly above or diagonally above the second feature, or simply indicates that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature includes the first feature being directly below or diagonally below the second feature, or simply indicates that the first feature is at a lower horizontal level than the second feature.

[0055] Please see Figure 1Three independent glass shake flasks or cell culture flasks are placed on the lab bench or in the incubator. Each flask has a special cap with multiple holes (such as a three- or four-hole cap). A bunch of silicone tubes of varying lengths emerge from the holes in each cap. These tubes connect like a spider web to several multi-channel peristaltic pumps nearby. At the other end of the peristaltic pumps, some tubing connects to a large fresh culture medium reservoir, and other tubing connects to a waste liquid collection bottle. The reservoir, waste liquid bottle, peristaltic pumps, and culture flasks are scattered in different locations on the lab bench or incubator. The following will describe this in detail from three aspects: structure, spatial connection, and process, and analyze its inherent defects.

[0056] First, the core structural components of this temporary assembly system include:

[0057] Culture containers: typically standard laboratory glass or disposable plastic shake flasks, serum bottles or special cell culture flasks (Spinner Flasks).

[0058] Multi-channel caps / stoppers: To allow for the entry and exit of liquids and gases, silicone stoppers or custom-made screw caps with 2 to 4 channels are typically used. Each channel is used to insert a tube or a sterile filter.

[0059] Piping system: mainly medical or food-grade silicone tubing, Teflon tubing and other flexible tubing.

[0060] Fluid drive devices: The most common type is the multi-channel peristaltic pump, which delivers liquid by controlling the pump's speed and start / stop.

[0061] External containers: including storage bottles for storing fresh culture medium and waste bottles for collecting waste liquid.

[0062] Gas exchange device: Typically, a 0.22µm sterile air filter is installed in one passage of the bottle cap to allow for gas exchange between the inside and outside while blocking microorganisms.

[0063] Secondly, the positions and connections between the components are as follows:

[0064] Locational relationship: The components are spatially dispersed and non-integrated. Culture flasks are placed in incubators or shakers; peristaltic pumps, storage bottles, and waste bottles are placed outside or inside the incubator depending on convenience, resulting in a large footprint and a chaotic layout.

[0065] Connection relationships:

[0066] Internal connections: Each culture flask cap has at least three tubes: an inlet tube (usually extending below the liquid surface for feeding), an outlet tube (which can be an venting / sampling tube above the liquid surface or a waste discharge tube below the liquid surface), and a vent tube connected to a sterile filter.

[0067] External connections: The inlet tube of each culture flask needs to be connected to a channel of the peristaltic pump via a separate tubing, and then from that channel to the main reservoir. Similarly, the outlet tube of each culture flask is connected to another channel of the peristaltic pump via another separate tubing (or directly using gravity / pressure), and then to the main waste bottle.

[0068] Result: This connection method leads to an extremely complex and chaotic tubing system. With N culture flasks, there are typically at least 2N long tubing lines crisscrossing between the flasks, pumps, and storage bottles, forming a so-called "tubing jungle." These tubing lines lack effective organization and fixation, making them highly susceptible to accidental stretching or crushing.

[0069] Finally, the process is as follows:

[0070] Preparation phase: Operators need to prepare each culture flask, cap, and all connecting tubing separately. Components including caps and tubing need to be assembled and sterilized individually (usually by autoclaving).

[0071] Assembly stage: After sterilization, all independent tubing systems are aseptically connected in a sterile operating room. For example, the inlet end of the culture flask assembly is aseptically connected to the tubing extending from the peristaltic pump. This is a high-risk operation, highly susceptible to contamination.

[0072] Operation phase: The peristaltic pump is used to replenish or drain specific culture flasks. When an operation is required on a particular flask, the operator needs to locate and configure the corresponding pump tubing and valve (if applicable) within the complex piping system.

[0073] The traditional independent culture flask method is the most basic, which uses multiple independent culture flasks, petri dishes, or shake flasks, each inoculated with samples and placed in an incubator or shaker. When operations such as adding materials, taking samples, or changing liquids are required, the operator needs to remove each culture container from its culture environment and open the cap of each flask or dish one by one. This method has the following significant disadvantages: It is cumbersome and inefficient: especially when there are many samples, operating each one individually is time-consuming and labor-intensive, greatly reducing experimental efficiency; it has a high risk of contamination: each time the culture container is opened, its internal environment is exposed to the outside environment, increasing the risk of introducing airborne microorganisms and contaminants brought in by the operator, leading to culture failure or unreliable experimental results; it has a risk of cross-contamination: when handling multiple samples, improper operation (such as mixing equipment or generating aerosols) can easily lead to cross-contamination between samples; and it is difficult to achieve precise synchronous control: because it is a manual, time-sharing operation, it is difficult to ensure that all samples are precisely exchanged or processed at the same time.

[0074] Partially Integrated or Automated Culture Systems: Some systems on the market attempt to integrate multiple culture units, such as micro-bioreactor arrays or automated cell culture workstations. While these systems offer improvements in automation and control precision, they often suffer from the following drawbacks: Complex Structure and High Cost: These systems typically include sophisticated fluid control components, sensors, and control software, resulting in complex equipment structures, high purchase and maintenance costs, and unsuitability for all laboratories; Insufficient Flexibility and Independence in Liquid Management: Even integrated systems may present design compromises or operational inconveniences in ensuring complete independence, sealing, and ease of liquid supply, discharge, and sampling for each culture unit. For example, piping connections may remain cumbersome, or it may be difficult to operate a single unit flexibly without affecting other units; Difficult Cleaning and Sterilization: Complex piping and cavity structures can pose challenges to thorough cleaning and sterilization.

[0075] In view of the problems of disordered, numerous, difficult to manage, and easily confused operation and accidental interference in the existing pipeline system, this application aims to provide a structure that can manage the liquid exchange pipelines required for multiple culture bottles in an orderly and integrated manner, so as to make the operation interface clear, the pipeline connection stable, and avoid misoperation or connection failure caused by pipeline disorder.

[0076] In view of the problems of poor overall system sealing and high risk of contamination caused by numerous exposed pipe connection points in the prior art, this application aims to provide a structure that can enhance the overall sealing of the system, especially to effectively protect and isolate the interface part of liquid exchange, thereby significantly reducing the risk of external environmental pollutants or cross-contaminants entering the culture system and improving the aseptic assurance level of the culture process.

[0077] In view of the problems of low space utilization caused by the dispersed layout of components in the existing technology, and the sharp increase in system complexity and difficulty in expansion when the number of culture units is increased, this application aims to provide a multi-bottle culture scheme with compact structure, high space utilization and easy expansion, so that more samples can be cultured independently in a limited space, and the increase of culture units will not lead to a catastrophic deterioration of pipeline management.

[0078] To address the problems of lack of standardization, low integration, and difficulty in ensuring operational consistency and experimental repeatability in existing temporary assembly systems, this application aims to provide an integrated and standardized structural design that simplifies the construction and operation process of multi-bottle independent culture systems, and improves the reliability of the system and the repeatability of experimental results.

[0079] For details, please refer to Figure 2 , Figure 3 , Figure 4This application provides a conveniently disassembled loop cell culture device, which includes:

[0080] Cell culture chamber 1 is equipped with a sealed door 111;

[0081] The shaking device 11 is disposed inside the cell culture chamber 1;

[0082] An integrated monitoring device 4 is disposed inside the cell culture chamber 1 and on the shaker device 11;

[0083] At least one culture flask 5 is disposed inside the cell culture chamber 1 and located above the integrated monitoring device 4;

[0084] An intermediate partition 2 is disposed at the top of the cell culture chamber 1; the intermediate partition 2 is provided with a sealing communication hole 21;

[0085] The liquid supply sampling chamber 3 is located above the cell culture chamber 1; the liquid supply sampling chamber 3 and the cell culture chamber 1 are connected through the sealing communication hole 21;

[0086] A replenishment circuit is provided inside the liquid supply and sampling chamber 3, and the replenishment circuit is connected to the culture bottle 5 through the sealing communication hole 21.

[0087] An alkali replenishment circuit is located inside the liquid supply sampling chamber 3, and the alkali replenishment circuit passes through the sealing communication hole 21 and is connected to the culture bottle 5.

[0088] A sampling circuit is provided inside the liquid supply sampling chamber 3, and the sampling circuit passes through the sealing communication hole 21 and is connected to the culture bottle 5.

[0089] The control host 9 is located outside the liquid supply and sampling chamber 3. The control host 9 is connected to the integrated monitoring device 4, the liquid replenishment circuit, the alkali replenishment circuit and the sampling circuit.

[0090] The cell culture chamber 1 is used to contain and protect the culture flasks 5, and can isolate the external environment to a certain extent, reducing airflow disturbance during operation. The cell culture chamber 1 includes a base and side plates, and the side plates may be equipped with observation windows or removable maintenance doors.

[0091] The shaker device 11 is preferably a magnetic shaker, which stirs the liquid by shaking the contents of the flask, ensuring uniform composition of the culture medium, promoting gas exchange and mass transfer, and preventing cell sedimentation. If the culture is sensitive to shear force or has special requirements, in addition to shaking, magnetic stirring or airlift stirring (which requires gas introduction) can also be considered.

[0092] The culture flask 5 is placed on the shaker device 11 of the base. The side plate is structurally connected to the base and the middle partition 2 above it to form a closed or semi-closed cell culture chamber 1.

[0093] The intermediate partition 2 clearly divides the entire device into two functional areas: the lower culture area and the upper fluid control and piping area. It supports all the fluid control components in the upper part, forming the "top cover" of the lower cell culture chamber 1, and also serves as the mounting base for all fluid control components in the upper liquid supply and sampling chamber 3. The intermediate partition 2 can precisely position each culture flask 5. The bottle mouth / sealing cap 51 of the culture flask 5 can be connected to the sealing communication hole 21 using a sealing cap 51 assembly and piping to ensure spatial isolation between the lower cell culture chamber 1 and the upper liquid supply and sampling chamber 3, preventing direct interference from upper operations to the lower culture environment, and laying the foundation for the relative sealing of the lower cell culture chamber 1. The intermediate partition 2 significantly reduces the risk of contamination introduced by the external environment or cross-operations, improving the success rate of culture and the reliability of data.

[0094] Material and processing of intermediate partition 2: It can be stainless steel, aluminum alloy, or corrosion-resistant engineering plastic, and can be manufactured by CNC machining, 3D printing, etc.

[0095] The upper liquid supply and sampling chamber 3 enables independent, precise, and automated control of the liquid in each culture bottle 5; it shortens the pipeline length (vertical direction) from the control element to the culture bottle 5, reducing dead volume; and all control and connection points are centralized, facilitating automation and troubleshooting.

[0096] The replenishment peristaltic pump 6, the alkali replenishment peristaltic pump 7, and the sampling peristaltic pump 8 utilize pneumatic pinch valves, which are particularly suitable for applications requiring avoidance of direct fluid contact with the valve body material, thus ensuring greater sterility of the flow path. The replenishment peristaltic pump 6 and the alkali replenishment peristaltic pump 7 provide independent, precisely controlled liquid delivery to each culture flask 5, enabling precise replenishment of fresh culture medium and addition of inducers / nutrients. The sampling peristaltic pump 8 extracts culture samples. The replenishment peristaltic pump 6, the alkali replenishment peristaltic pump 7, and the sampling peristaltic pump 8 employ a valve array to control the flow direction and on / off state of the fluid in the pipeline. Through programmed valve control, automatic switching and independent operation of different flow paths for each culture flask 5, such as liquid inlet, liquid outlet, and sampling, can be achieved.

[0097] The liquid replenishment peristaltic pump 6 and the alkali replenishment peristaltic pump 7 are connected to the liquid storage / buffer unit and can be used to temporarily store reagents to be added, fresh culture medium, or as a buffer before waste liquid collection, or as a bubble trap in the pipeline system, etc.

[0098] The piping system connects the culture flask 5, the replenishment peristaltic pump 6, the alkali replenishment peristaltic pump 7, the sampling peristaltic pump 8, and the storage / buffer unit, forming a complete and independently controllable liquid exchange circuit. One end of these pipes communicates with the inside of the culture flask 5 through the socket on the cap, and the other end is connected to the valve and the peristaltic pump.

[0099] Please see Figure 9 The culture flask 5 has a sealing cap 51 at its mouth, and the sealing cap 51 has three connecting tubes 52, which are respectively connected to the liquid replenishment circuit, the alkali replenishment circuit, and the sampling circuit. The culture flask 5 serves as a direct container for biological culture, used to hold culture medium and biological samples. Each culture flask 5 is equipped with a sealing cap 51 with multiple ports for connecting inlet, outlet, and venting tubes. The sealing cap 51 centralizes the tubing interfaces, ensuring a sealed protection at the critical connection point for liquid exchange (the sealing cap 51). The culture flask 5 can be a standard glass or disposable plastic conical flask, serum bottle, or a specially customized culture container with more interfaces or built-in sensors.

[0100] Please see Figure 3 The fluid replenishment circuit includes:

[0101] At least one replenishment bottle 12 is fixed inside the liquid supply and sampling chamber 3;

[0102] At least one replenishment peristaltic pump 6 is provided in the liquid supply and sampling chamber 3. The replenishment peristaltic pump 6 is used to draw out the culture medium in the replenishment bottle 12 and replenish it into the culture bottle 5.

[0103] The alkali replenishment circuit includes:

[0104] At least one alkali replenishment bottle 13 is fixed inside the liquid supply sampling chamber 3;

[0105] At least one alkali replenishment peristaltic pump 7 is provided in the liquid supply sampling chamber 3. The alkali replenishment peristaltic pump 7 is used to draw alkali solution from the alkali replenishment bottle 13 and replenish it to the culture bottle 5.

[0106] The sampling circuit includes:

[0107] At least one sampling peristaltic pump 8 is disposed in the liquid supply sampling chamber 3, and the sampling peristaltic pump 8 is used to extract cell fluid from the culture flask 5;

[0108] The sampling and dispensing device 10 is connected to the sampling peristaltic pump 8, and the sampling and dispensing device 10 dispenses the cell fluid extracted by the sampling peristaltic pump 8 into the sampling tube.

[0109] Please see Figure 3 , Figure 4The liquid supply sampling chamber 3 is also provided with a culture medium bottle fixing seat 31, an alkali bottle fixing seat 32 and a liquid tube fixing bracket 33. The culture medium bottle fixing seat 31 is located near the replenishment peristaltic pump 6. The replenishment bottle 12 is fixed on the culture medium bottle fixing seat 31. The alkali bottle fixing seat 32 is located near the alkali replenishment peristaltic pump 7. The alkali replenishment bottle 13 is fixed on the alkali bottle fixing seat 32. The liquid tube fixing bracket 33 is provided with a liquid tube fixing hole.

[0110] Please see Figure 4 The easily detachable loop-type cell culture device further includes a cooling module 14, which is disposed within the liquid supply and sampling chamber 3 and corresponding to the culture medium bottle fixing base 31. The cooling module 14 can cool the culture medium bottle to adapt to the temperature required for cell culture. The cooling module 14 includes a fan and cooling elements.

[0111] Please see Figure 8 The culture flask 5 is provided in four parts, and the four culture flasks 5 are evenly distributed on the integrated monitoring device 4; the alkali replenishment bottle 13 and the liquid replenishment peristaltic pump 6 are connected to the culture flask 5 in a one-to-one correspondence, the alkali replenishment bottle 13 and the alkali replenishment peristaltic pump 7 are connected to the culture flask 5 in a one-to-one correspondence, and the sampling peristaltic pump 8 is connected to the culture flask 5 in a one-to-one correspondence.

[0112] Please see Figure 4 The liquid supply sampling chamber 3 is further provided with a fixing bracket 34, which extends to the outer surface of the liquid supply sampling chamber 3. The control host 9 is fixed on the fixing bracket 34 and is flush with the outer surface of the liquid supply sampling chamber 3. The fixing bracket 34 is used to fix the control host 9, facilitating the operation of the control host 9.

[0113] Please see Figure 5 The sampling and dispensing device 10 includes:

[0114] Liquid separation module 101, the liquid separation module 101 is provided with an inlet and multiple outlets, the inlet is connected to the sampling peristaltic pump 8, and the outlets are located on the lower surface of the liquid separation module 101;

[0115] The sampling box 102 is removable and installed in the liquid supply sampling chamber 3. The sampling box 102 is provided with a sampling slot 103. The sampling slot 103 is arranged along the extension direction of the liquid distribution module 101 and is located below the liquid outlet.

[0116] A sampling tube is inserted into the sampling slot 103, and the sampling tube receives the cell fluid flowing out from the outlet.

[0117] Please see Figure 5 The sampling and dispensing device 10 further includes:

[0118] Waste liquid pool 104 is located inside the sampling box 102, arranged along the extending direction of the liquid separation module 101, and parallel to the sampling slot 103;

[0119] A transverse drive structure 105 is connected to the liquid distribution module 101 and controls the liquid distribution module 101 to move along the bottom plate direction parallel to the liquid supply sampling chamber 3. The transverse drive structure 105 drives the liquid distribution module 101 to move between the sampling slot 103 and the waste liquid pool 104.

[0120] Please see Figure 5 , Figure 6 , Figure 7 The cell culture chamber 1 has a guide groove 112 at the top corresponding to the sealed communication hole 21. The intermediate partition 2 includes a liquid tube clamping device 22. The liquid tube clamping device 22 is located on the bottom plate of the liquid supply and sampling chamber 3 and is located on one side of the sealed communication hole 21. The other side of the sealed communication hole 21 has a first liquid tube contour groove 211. The liquid tube clamping device 22 has a second liquid tube contour groove 212 corresponding to the first liquid tube contour groove 211. The first liquid tube contour groove 211 and the second liquid tube contour groove 212 form the sealed communication hole 21. The liquid tube clamping device 22 presses and seals the liquid tube in the sealed communication hole 21 by pushing and pulling.

[0121] Please see Figure 5 , Figure 6 , Figure 7 The liquid tube clamping device 22 includes a rotary knob 221, a rotating shaft 222, a cam 223, a push plate 224, and a reset elastic element 225. The rotating shaft 222 is perpendicular to the bottom plate of the liquid supply sampling chamber 3. The cam 223 is connected to the middle of the rotating shaft 222. The rotary knob 221 is connected to the top of the rotating shaft 222. The push plate 224 abuts against the cam 223. The reset elastic element 225 connects the bottom plate of the liquid supply sampling chamber 3 to the push plate 224. The push plate 224 has a second liquid tube contour groove 212 on the side away from the reset elastic element 225, and the liquid tube is clamped between the second liquid tube contour groove 212 and the first liquid tube contour groove 211.

[0122] The liquid tube clamping device 22 separates and seals the cell culture chamber 1 and the liquid supply and sampling chamber 3 through the intermediate partition 2, which can form a relatively closed space in the lower cell culture chamber 1. This helps to maintain the stability of the environment around the culture bottle 5 and reduce the direct intrusion of external dust or aerosols.

[0123] The integrated monitoring device 4 includes an oxygen concentration monitoring module, a pH monitoring module, a turbidity monitoring module, a cell counting module, a temperature monitoring module, and a carbon dioxide concentration monitoring module. This structure facilitates the integration of sensors and corresponding control systems, such as pH, DO (dissolved oxygen), and temperature sensors, to achieve a higher degree of automated monitoring and feedback regulation of the culture process.

[0124] Furthermore, the easily detachable loop cell culture device also includes an audible and visual alarm system connected to the control host 9. The audible and visual alarm system can issue an audible and visual alarm signal when the integrated monitoring device 4 detects abnormal data.

[0125] The easily disassembled loop cell culture device provided in this application embodiment includes a cell culture chamber 1, an intermediate partition 2, and a liquid supply and sampling chamber 3. This achieves a sealed environment for the cell culture process, avoiding the risk of contamination. Furthermore, an integrated monitoring device 4 is installed in the cell culture chamber 1 to monitor the culture flask 5 placed above it. A liquid replenishment peristaltic pump 6, an alkali replenishment peristaltic pump 7, and a sampling peristaltic pump 8 are installed in the liquid supply and sampling chamber 3. Together with the control host 9, the device enables real-time replenishment of culture medium or alkali based on the monitored conditions in the culture flask 5, and allows for sampling of cell fluid from the culture flask 5 at appropriate times. This fully automates the process. Moreover, the liquid replenishment loop, alkali replenishment loop, sampling loop, and culture flask 5 are easy to disassemble and replace, improving the success rate and efficiency of cell culture, reducing manual intervention, and lowering the risk of contamination.

[0126] In the above embodiments, the descriptions of each embodiment have different focuses. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0127] The above provides a detailed description of a conveniently detachable loop cell culture device provided in the embodiments of this application. Specific examples have been used to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the technical solutions and core ideas of this application. Those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of this application.

Claims

1. A conveniently detachable loop-type cell culture device, characterized in that, The easily detachable loop cell culture device includes: The cell culture chamber is equipped with a sealed door; The shaking device is located inside the cell culture chamber; An integrated monitoring device is disposed within the cell culture chamber and is mounted on the shaker device; At least one culture flask is disposed within the cell culture chamber and located above the integrated monitoring device; An intermediate partition is disposed at the top of the cell culture chamber; the intermediate partition is provided with a sealing communication hole; A liquid supply and sampling chamber is located above the cell culture chamber; the liquid supply and sampling chamber is connected to the cell culture chamber through the sealed communication hole; A replenishment circuit is provided in the liquid supply and sampling chamber, and the replenishment circuit is connected to the culture flask through the sealed communication hole; An alkali replenishment circuit is provided in the liquid supply sampling chamber, and the alkali replenishment circuit is connected to the culture flask through the sealed communication hole; A sampling circuit is provided in the liquid supply sampling chamber, and the sampling circuit is connected to the culture flask through the sealed communication hole; The control host is located outside the liquid supply and sampling chamber, and the control host is connected to the integrated monitoring device, the liquid replenishment circuit, the alkali replenishment circuit and the sampling circuit.

2. The easily detachable loop-type cell culture device as described in claim 1, characterized in that, The culture flask is equipped with a sealing cap at its mouth, and the sealing cap has three connecting tubes, which are respectively connected to the liquid replenishment circuit, the alkali replenishment circuit, and the sampling circuit.

3. The easily disassembled loop-type cell culture device as described in claim 1, characterized in that, The fluid replenishment circuit includes: At least one replenishment bottle is fixed inside the liquid supply and sampling chamber; At least one rehydration peristaltic pump is provided in the liquid supply and sampling chamber. The rehydration peristaltic pump is used to draw culture medium from the rehydration bottle and replenish it into the culture bottle. The alkali replenishment circuit includes: At least one alkali replenishment bottle is fixed inside the liquid supply sampling chamber; At least one alkali replenishment peristaltic pump is installed in the liquid supply sampling chamber. The alkali replenishment peristaltic pump is used to draw alkali solution from the alkali replenishment bottle and replenish it to the culture bottle. The sampling circuit includes: At least one sampling peristaltic pump is disposed in the liquid supply sampling chamber, and the sampling peristaltic pump is used to draw cell fluid from the culture flask; A sampling and dispensing device is connected to the sampling peristaltic pump, and the sampling and dispensing device distributes the cell fluid extracted by the sampling peristaltic pump into the sampling tube.

4. The easily disassembled loop-type cell culture device as described in claim 3, characterized in that, The liquid supply sampling chamber is also equipped with a culture medium bottle holder, an alkali bottle holder, and a liquid tube holder. The culture medium bottle holder is located adjacent to the replenishment peristaltic pump, and the replenishment bottle is fixed on the culture medium bottle holder. The alkali bottle holder is located adjacent to the alkali replenishment peristaltic pump, and the alkali replenishment bottle is fixed on the alkali bottle holder. The liquid tube holder is provided with a liquid tube fixing hole.

5. The easily detachable loop cell culture device as described in claim 4, characterized in that, The easily detachable loop cell culture device also includes: A cooling module is located within the liquid supply and sampling chamber and is positioned corresponding to the culture medium bottle fixing seat.

6. The easily detachable loop cell culture device as described in claim 3, characterized in that, The culture flasks are provided in four parts, and the four culture flasks are evenly distributed on the integrated monitoring device; the alkali replenishment bottle and the liquid replenishment peristaltic pump are connected to the culture flasks one by one, and the sampling peristaltic pump is connected to the culture flasks one by one.

7. The easily detachable loop cell culture device as described in claim 3, characterized in that, The sampling and dispensing device includes: The liquid separation module is provided with an inlet and multiple outlets. The inlet is connected to the sampling peristaltic pump, and the outlets are located on the lower surface of the liquid separation module. A sampling box is removable and installed in the liquid supply sampling chamber. The sampling box is provided with a sampling slot, which is arranged along the extension direction of the liquid dispensing module and is located below the liquid outlet. A sampling tube is inserted into the sampling slot and receives cell fluid flowing out from the outlet.

8. The easily detachable loop cell culture device as described in claim 7, characterized in that, The sampling and dispensing device further includes: A waste liquid pool is located inside the sampling box, arranged along the extension direction of the liquid separation module, and parallel to the sampling slot; A transverse drive structure is connected to the liquid dispensing module to control the liquid dispensing module to move along the bottom plate direction parallel to the liquid supply sampling chamber. The transverse drive structure drives the liquid dispensing module to move between the sampling slot and the waste liquid pool.

9. The easily detachable loop cell culture device as described in claim 1, characterized in that, The top of the cell culture chamber is provided with a guide groove corresponding to the sealed communication hole. The intermediate partition includes a liquid tube clamping device. The liquid tube clamping device is located on the bottom plate of the liquid supply and sampling chamber and is located on one side of the sealed communication hole. A first liquid tube contour groove is provided on the other side of the sealed communication hole. The liquid tube clamping device is provided with a second liquid tube contour groove corresponding to the first liquid tube contour groove. The first liquid tube contour groove and the second liquid tube contour groove form the sealed communication hole. The liquid tube clamping device presses and seals the liquid tube in the sealed communication hole by pushing and pulling.

10. The easily detachable loop-type cell culture device as described in claim 9, characterized in that, The liquid tube clamping device includes a rotary knob, a rotating shaft, a cam, a push plate, and a reset elastic element. The rotating shaft is perpendicular to the bottom plate of the liquid supply sampling chamber. The cam is connected to the middle of the rotating shaft. The rotary knob is connected to the top of the rotating shaft. The push plate abuts against the cam. The reset elastic element connects the bottom plate of the liquid supply sampling chamber to the push plate. The push plate has a second liquid tube conforming groove on the side opposite to the reset elastic element, and the liquid tube is clamped between the second liquid tube conforming groove and the first liquid tube conforming groove.

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