Sterile culture method and sterile culture system

Abstract

The present invention relates to the technical field of biopharmaceutical apparatuses, in particular to a sterile culture method and a sterile culture system. The sterile culture system comprises a cell preparation device, an incubator, and a cell culture device; the cell preparation device comprises a preparation chamber, wherein a first sealing valve is arranged on a side wall of the preparation chamber; the incubator has a culture cavity, wherein a second sealing valve is arranged on a side wall of the culture cavity, and the second sealing valve is configured to work with the first sealing valve such that the incubator is hermetically docked with or separated from the cell preparation device; and the cell culture device is used for accommodating the incubator, supplying power and a gas to the incubator, and controlling a cell growth environment in the incubator. Because cells grow in a sterile environment all the time and the pollution risk is relatively low, no frequent sterilization is required. Normal growth of cells is avoided from being affected by frequent sterilization, and the culture time is saved. Therefore, the cell preparation and culture efficiency is relatively high.

Description

Aseptic culture method and aseptic culture system Technical Field

[0001] The present invention relates to the technical field of biopharmaceutical equipment, and in particular to an aseptic culture method and an aseptic culture system. Background Art

[0002] CGT (cell and gene therapy) products are cutting-edge technologies in the pharmaceutical field, with high added value and huge market potential. They are part of the medicines that cure diseases and save lives. Like conventional chemical drugs, they must also ensure the sterility of the production process. Once contaminated during the production process, patients will lose the best treatment opportunity due to the inability to obtain drugs in time. In severe cases, the patient may lose his or her life, and CGT pharmaceutical companies will suffer significant losses.

[0003] In addition, CGT drug culture requires strict time control. If good time control is not carried out during the preparation and culture process, it will affect the cell growth condition, and the cell culture condition will directly affect the treatment effect of patients receiving cell therapy. CGT drugs are biologically active drugs, and they need to be observed and replaced with culture media at any time during the culture process. The traditional way is to transfer the cell drugs to a conventional incubator for culture after they are prepared in the cell preparation device. When observation and culture media replacement are required during the culture process, they are transferred to the cell preparation device for processing. After completion, they are transferred to the incubator for continued culture, and the operation is repeated repeatedly.

[0004] During the reciprocating movement of cell-derived drugs from the isolation system to the incubator, the cell culture container will pass through the laboratory's B / C / D low-level environments, and the probability of cell contamination is extremely high. The personnel performing the transfer operation need to have very professional skills, and the operator's condition will directly affect the risk of cell contamination during the transfer process. The entire culture process in conventional incubators is in a low-level environment, and the cell culture process has a high risk of contamination. In addition, the culture container containing cells is frequently sterilized, and the sterilant has the risk of penetrating the cell container and killing the cells. Therefore, the cell culture process of existing sterile culture methods has problems such as high contamination risk and low cell preparation and culture efficiency.

[0005] Summary of the Invention

[0006] The purpose of the present invention is to provide a sterile culture method and a sterile culture system to solve the problems of high contamination risk and low cell preparation and culture efficiency in the process of culturing cells in the prior art.

[0007] In order to solve the above technical problems, the present invention provides a sterile culture method.

[0008] The aseptic culture method of the present invention comprises:

[0009] Providing a cell preparation device and an incubator;

[0010] Sealing and docking the incubator with the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device are in communication with each other;

[0011] sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator, and after sterilization, sealing and separating the incubator from the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device remain sealed during separation;

[0012] preparing cells in a preparation chamber of the cell preparation device;

[0013] Sealing and docking the incubator with the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device are in communication with each other, and transferring cells from the preparation chamber of the cell preparation device to the culture chamber of the incubator;

[0014] Seal and separate the incubator from the cell preparation device, so that the culture chamber of the incubator and the preparation chamber of the cell preparation device remain sealed during separation;

[0015] A cell culture device is provided, and the cell culture environment in the incubator is controlled by the cell culture device to culture the cells in the incubator.

[0016] Furthermore, the cell preparation device and the incubator are provided with mutually cooperating sealing docking structures;

[0017] The sealing docking structure includes an RTP main valve and an RTP auxiliary valve, and the RTP main valve and the RTP auxiliary valve form an RTP valve when they are buckled together;

[0018] The process of sealingly docking the incubator with the cell preparation device includes: rotating the RTP main valve so that the RTP main valve and the RTP auxiliary valve are fastened together to form an RTP valve; opening the RTP valve to achieve sealed docking between the incubator and the cell preparation device;

[0019] The process of sealing and separating the incubator from the cell preparation device includes: closing the RTP valve; and rotating the RTP main valve to separate the RTP main valve and the RTP auxiliary valve to achieve sealed separation of the incubator from the cell preparation device.

[0020] Furthermore, the process of sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator includes:

[0021] introducing sterile gas into the cell preparation device and / or the incubator, and allowing the sterile gas to diffuse and circulate in the incubator and the cell preparation device;

[0022] After the sterilization is completed, the sterilization gas in the preparation chamber and the culture chamber is exhausted.

[0023] Furthermore, while sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator, the sterilizing filter of the incubator is sterilized using the sterilizing gas, and the sterilizing filter is connected to the culture chamber through a gas pipeline.

[0024] The present invention also provides a sterile culture system, comprising a cell preparation device, an incubator, and a cell culture device;

[0025] The cell preparation device includes a preparation chamber, and a first sealing valve is provided on the side wall of the preparation chamber;

[0026] The incubator has a culture chamber, and a second sealing valve is provided on a side wall of the culture chamber, and the second sealing valve is used to cooperate with the first sealing valve to achieve sealed docking or sealed separation between the incubator and the cell preparation device;

[0027] The cell culture device is used to accommodate an incubator, supply power and gas to the incubator, and control the cell growth environment in the incubator.

[0028] Furthermore, one of the first sealing valve and the second sealing valve is an RTP main valve, and the other is an RTP auxiliary valve. When the RTP main valve and the RTP auxiliary valve are buckled together, they form an RTP valve.

[0029] Furthermore, the edge of the RTP main valve is provided with a first cylinder and a second cylinder for rotating the RTP main valve, and the extension and contraction directions of the first cylinder and the second cylinder are set to be parallel and opposite;

[0030] The RTP main valve or the RTP auxiliary valve is provided with a switch gate mechanism, and the switch gate mechanism is used to open or close the RTP valve.

[0031] Furthermore, the cell preparation device further includes a sterilization device for generating sterilization gas, and an output port of the sterilization device is connected to the preparation chamber.

[0032] Furthermore, the cell preparation device also includes a circulation fan, and the incubator is provided with a turbulent fan. The circulation fan and the turbulent fan cooperate to fully diffuse the sterilization gas in the preparation chamber and the culture chamber.

[0033] Furthermore, the cell preparation device also includes an exhaust component, which is used to replace the sterilization gas and discharge it after the sterilization is completed.

[0034] Furthermore, it also includes: a vacuum pump and a suction pipeline;

[0035] A sterilizing filter is provided in the incubator, and the sterilizing filter is connected to the culture chamber through a gas pipeline;

[0036] The suction pipeline is connected to the sterilizing filter, and the vacuum pump is arranged on the suction pipeline for sucking the sterilizing gas in the incubator to sterilize the sterilizing filter.

[0037] Furthermore, it also includes: an air blowing pipeline, which is connected to the sterilizing filter and is used to blow air into the sterilizing filter to discharge the sterilizing gas in the sterilizing filter after sterilization is completed.

[0038] Furthermore, the cell culture device has a plurality of accommodating spaces arranged in an array, and the incubators placed in the accommodating spaces are all centrally controlled by the control unit of the cell culture device.

[0039] Furthermore, a transfer trolley is included, and the transfer trolley is used to transfer the incubator.

[0040] Compared with the prior art, the present invention has at least the following beneficial effects:

[0041] Before use, the incubator and the cell preparation device are sealed and docked together, thereby connecting the gas environment of the culture chamber with the gas environment of the preparation chamber. Then, the two are sterilized and, after sterilization, are sealed and separated. Then, cells are prepared in the preparation chamber. After the cell preparation is completed, the second sealing valve of the incubator is matched with the first sealing valve of the cell preparation device, thereby sealing and docking the incubator and the cell preparation device, thereby connecting the gas environment of the culture chamber with the gas environment of the preparation chamber. In this sealed sterile environment, the cells are transferred to the incubator. Then, the incubator and the cell preparation device are sealed and separated, and the incubator is transferred to the cell culture device to culture the cells.

[0042] Because the cell preparation device and incubator are sterilized before cell preparation, the prepared cells grow in a sterile environment. Furthermore, because the incubator and cell preparation device are sealed and docked before the cells are transferred to the incubator, and the cells are transferred to the incubator in a sealed environment, the cells always grow in a sterile environment, with a low risk of contamination. Therefore, frequent sterilization is not required, which not only avoids the impact of frequent sterilization on normal cell growth, but also saves culture time, resulting in higher cell preparation and culture efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG1 is a schematic structural diagram of an embodiment of a sterile culture system of the present invention;

[0044] FIG2 is a schematic structural diagram of the cell preparation device and the incubator in FIG1 after being docked and sterilized together;

[0045] FIG3 is a schematic diagram of the structure of the cell preparation device and the incubator in FIG1 after partially hiding the structure when docking;

[0046] FIG4 is a schematic structural diagram of the cell preparation device in FIG1 with part of the structure hidden to show the RTP main valve;

[0047] FIG5 is a partial enlarged view of point A in FIG4 ;

[0048] Figure numerals: 100, cell preparation device; 110, culture chamber; 120, support frame; 130, limiter; 141, main valve body; 142, main valve plate; 143, first rotary drive cylinder; 144, second rotary drive cylinder; 145, drive motor; 146, connecting rod assembly; 147, latch shaft; 148, hinge movable leaf; 149, support block; 150, latch seat; 151, RTP auxiliary valve; 161, control unit; 162, first in-position signal sensor; 163, second in-position signal sensor; 164, third in-position signal sensor; 165, fourth in-position signal sensor; 166, power socket; 171, liquid storage bottle; 172, delivery pump; 173, delivery pipeline; 174, gas generating device; 175, circulation fan; 176, exhaust assembly; 200. Incubator; 210. Turbine fan; 221. First sterilizing filter; 222. Second sterilizing filter; 223. Suction line; 225. First, second, two-way solenoid valve; 226. Vacuum pump; 227. Air blowing line; 228. Second, second, two-way solenoid valve; 229. Compressed air flow regulating valve; 300. Cell culture apparatus; 400. Transfer cart. DETAILED DESCRIPTION

[0049] The following description of an aseptic culture method and aseptic culture system of the present invention is provided in conjunction with schematic diagrams, which illustrate preferred embodiments of the present invention. It should be understood that those skilled in the art may modify the present invention described herein while still achieving the advantageous effects of the present invention. Therefore, the following description should be understood as being generally known to those skilled in the art and not intended to limit the present invention.

[0050] The serial numbers of the components herein, such as "first", "second", etc., are only used to distinguish the objects described and do not have any order or technical meaning. The "connection" and "coupling" mentioned in this application, unless otherwise specified, include direct and indirect connections (couplings). In the description of the present invention, it should be understood that the orientation or positional relationship indicated by the terms "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", etc. are based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0051] In the present invention, unless otherwise expressly specified or limited, when a first feature is "above" or "below" a second feature, it may mean that the first and second features are in direct contact, or that the first and second features are in indirect contact through an intermediary. Furthermore, when a first feature is "above," "above," or "above" a second feature, it may mean that the first feature is directly above or diagonally above the second feature, or simply means that the first feature is at a higher level than the second feature. When a first feature is "below," "below," or "below" a second feature, it may mean that the first feature is directly below or diagonally below the second feature, or simply means that the first feature is at a lower level than the second feature.

[0052] The present invention is described in more detail in the following paragraphs by way of example with reference to the accompanying drawings. The advantages and features of the present invention will become more apparent from the following description and claims. It should be noted that the drawings are greatly simplified and not to exact scale, and are provided solely for the purpose of assisting in the description of the embodiments of the present invention.

[0053] The following describes the aseptic culture method in the embodiment of the first aspect of the present invention in conjunction with the description.

[0054] In one embodiment, the aseptic culture method of this embodiment comprises:

[0055] S100: providing a cell preparation device and an incubator;

[0056] S200: Sealing and docking the incubator with the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device are in communication with each other;

[0057] S300: sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator, and after sterilization, sealing and separating the incubator from the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device remain sealed during separation;

[0058] S400: preparing cells in a preparation chamber of the cell preparation device;

[0059] S500: Sealing and docking the incubator with the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device are in communication with each other, and transferring cells from the preparation chamber of the cell preparation device to the culture chamber of the incubator;

[0060] S600: Sealing and separating the incubator and the cell preparation device, so that the culture chamber of the incubator and the preparation chamber of the cell preparation device remain sealed during separation;

[0061] S700: providing a cell culture device, and controlling the cell culture environment in the incubator by the cell culture device to culture the cells in the incubator.

[0062] In step S300, hydrogen peroxide gas may be used to sterilize the cell preparation device and the incubator. In other embodiments, other types of sterilizing gases may be used, and other sterilization methods may be used for sterilization, such as ultraviolet sterilization.

[0063] In step S500, the docking process needs to ensure sealing to avoid contamination from the external environment. After docking, the cell preparation device and the gas environment in the incubator are connected. Since both are sterile environments, the transfer process is also completed in a sterile process, and the risk of contamination is low.

[0064] Because the cell preparation device and incubator are sterilized before cell preparation, the prepared cells grow in a sterile environment. Furthermore, because the incubator and cell preparation device are sealed and docked before the cells are transferred to the incubator, and the cells are transferred to the incubator in a sealed environment, the cells always grow in a sterile environment, with a low risk of contamination. Therefore, frequent sterilization is not required, which not only avoids the impact of frequent sterilization on normal cell growth, but also saves culture time, resulting in higher cell preparation and culture efficiency.

[0065] In one embodiment, the cell preparation device and the incubator are provided with mutually cooperating sealing docking structures. The sealing docking structure includes an RTP (Rapid Transfer Ports) main valve and an RTP auxiliary valve, and the RTP main valve and the RTP auxiliary valve form an RTP valve when they are fastened together. After the RTP main valve and the RTP auxiliary valve are fastened together, the outer surface of the RTP main valve and the outer surface of the RTP auxiliary valve are sealed in the RTP valve. The outer surface refers to the side exposed to the external environment. If there are bacteria, they will be sealed between the RTP main valve and the RTP auxiliary valve. By opening the RTP valve formed by the RTP main valve and the RTP auxiliary valve, the two sterile chambers can be connected.

[0066] The process of achieving sealed docking between the incubator and the cell preparation device using the sealed docking structure includes: rotating the RTP main valve so that the RTP main valve and the RTP auxiliary valve are engaged to form an RTP valve; and opening the RTP valve to achieve sealed docking between the incubator and the cell preparation device.

[0067] The process of using the sealed docking structure to achieve sealed separation of the incubator and the cell preparation device includes: closing the RTP valve; and rotating the RTP main valve to separate the RTP main valve and the RTP auxiliary valve to achieve sealed separation of the incubator and the cell preparation device.

[0068] The RTP main valve and RTP auxiliary valve can automatically achieve sealed docking or sealed separation between the cell preparation device and the incubator, making cell transfer more convenient and requiring no specialized skills. General laboratory assistants can complete the entire transfer process with simple operations, resulting in low labor costs and a pollution-free transfer process. Preferably, the RTP main valve is located on the cell preparation device, and the RTP auxiliary valve is located on the incubator. Of course, in other embodiments, the RTP auxiliary valve can also be located on the cell preparation device, and the RTP main valve on the incubator.

[0069] In one embodiment, in order to sterilize the cell preparation device and the incubator, before preparing cells, the incubator is sealed and docked with the cell preparation device, and sterilizing gas is introduced into the cell preparation device and / or inside, so that the sterilizing gas diffuses and circulates in the incubator and the cell preparation device.

[0070] The process of sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator includes:

[0071] introducing sterile gas into the cell preparation device and / or the incubator, and allowing the sterile gas to diffuse and circulate in the incubator and the cell preparation device;

[0072] After the sterilization is completed, the sterilization gas in the preparation chamber and the culture chamber is exhausted.

[0073] The sterilizing gas is preferably hydrogen peroxide, which can be generated from a hydrogen peroxide liquid. The sterilizing gas can be introduced into the cell preparation device and then diffused and circulated using a turbulent fan or other turbulent device. Alternatively, the sterilizing gas can be introduced into the incubator first, or simultaneously introduced into the cell preparation device and the incubator. Furthermore, other types of sterilizing gases can be selected as needed.

[0074] To prevent contamination of the incubator by the external environment, a sterilizing filter is provided outside the culture chamber and is connected to the culture chamber via a gas conduit. In one embodiment, the sterilizing gas is used to sterilize the sterilizing filter of the incubator while the preparation chamber of the cell preparation device and the culture chamber of the incubator are sterilized.

[0075] The following describes the sterile culture system in the embodiment of the second aspect of the present invention in conjunction with Figures 1 to 5 of the specification.

[0076] As shown in FIG. 1 and FIG. 2 , in one embodiment, the sterile culture system of this embodiment includes a cell preparation device 100 , an incubator 200 and a cell culture device 300 .

[0077] The cell preparation device 100 includes a preparation chamber, and a first sealing valve is provided on the side wall of the preparation chamber.

[0078] The incubator 200 has a culture chamber 110 , and a second sealing valve is provided on the side wall of the culture chamber 110 . The second sealing valve is used to cooperate with the first sealing valve to achieve sealed docking or sealed separation of the incubator 200 and the cell preparation device 100 .

[0079] The cell culture device 300 is used to accommodate the incubator 200, supply power and air to the incubator, and control the cell growth environment in the incubator.

[0080] Before use, the incubator 200 and the cell preparation device 100 are first sealed and docked, thereby connecting the gas environment of the culture chamber 110 with the gas environment of the preparation chamber. Then, the two are sterilized and sealed and separated after sterilization. Then, cells are prepared in the preparation chamber. After the cell preparation is completed, the second sealing valve of the incubator 200 is matched with the first sealing valve of the cell preparation device 100, thereby sealing and docking the incubator 200 and the cell preparation device 100, thereby connecting the gas environment of the culture chamber 110 with the gas environment of the preparation chamber. In this sealed sterile environment, the cells are transferred to the incubator 200. Then, the incubator 200 and the cell preparation device 100 are sealed and separated, and the incubator 200 is transferred to the cell culture device 300 to culture the cells.

[0081] Because the cell preparation apparatus 100 and incubator 200 are sterilized before cell preparation, the prepared cells grow in a sterile environment. Furthermore, because the incubator 200 and cell preparation apparatus 100 are sealed and docked before the cells are transferred to the incubator 200, and the cells are transferred to the incubator 200 in a sealed environment, the cells always grow in a sterile environment, reducing the risk of contamination. Therefore, frequent sterilization is not required, which not only avoids the impact on normal cell growth caused by frequent sterilization, but also saves culture time, resulting in higher cell preparation and culture efficiency.

[0082] Specifically, the cell preparation device 100 also includes a support frame 120, which is used to support the incubator 200. A limiter 130 is provided on the support frame 120, and the limiter 130 is used to limit the position of the incubator 200 so as to firmly fix the incubator 200 on the support frame 120.

[0083] In order to achieve sterile transfer, a sterile operation window is provided on the side wall of the preparation chamber, and sterile operation gloves, such as rubber gloves, are provided at the sterile operation window. The sterile operation gloves are located in the preparation chamber and are sealed to the sterile operation window. The operator only needs to insert his hands into the sterile operation gloves to perform operations without opening the preparation chamber, such as transferring cells in the preparation chamber to the docked incubator 200, or transferring cells in the docked incubator 200 to the preparation chamber.

[0084] In one embodiment, as shown in Figures 3, 4, and 5, one of the first sealing valve and the second sealing valve is an RTP (Rapid Transfer Ports) main valve, and the other is an RTP secondary valve 151. Preferably, the first sealing valve is the RTP main valve, and the second sealing valve is the RTP secondary valve 151, that is, the RTP main valve is provided in the cell preparation apparatus 100, and the RTP secondary valve 151 is provided in the incubator 200.

[0085] When the RTP main valve and RTP auxiliary valve 151 are fastened together, the outer surface of the RTP main valve and the outer surface of the RTP auxiliary valve 151 are sealed within the RTP valve. The outer surface refers to the side exposed to the external environment. Any bacteria will be trapped between the RTP main valve and the RTP auxiliary valve 151. Opening the RTP valve, formed by the fastened RTP main valve and RTP auxiliary valve 151, connects the two sterile chambers.

[0086] The process of achieving sealed docking between the incubator and the cell preparation device using the sealed docking structure includes: rotating the RTP main valve so that the RTP main valve and the RTP auxiliary valve 151 are engaged to form an RTP valve; and opening the RTP valve to achieve sealed docking between the incubator and the cell preparation device.

[0087] The process of using the sealed docking structure to achieve sealed separation of the incubator and the cell preparation device includes: closing the RTP valve; and rotating the RTP main valve to separate the RTP main valve and the RTP auxiliary valve 151 to achieve sealed separation of the incubator and the cell preparation device.

[0088] The RTP main valve and RTP auxiliary valve 151 can automatically achieve sealed docking or sealed separation between the cell preparation device 100 and the incubator 200, making cell transfer more convenient and requiring no specialized skills. General laboratory assistants can complete the entire transfer process with simple operations, resulting in low labor costs. Of course, in other embodiments, the RTP auxiliary valve 151 can also serve as the first sealing valve, and the RTP main valve can serve as the second sealing valve.

[0089] Specifically, the RTP main valve includes a main valve body 141 and a main valve plate 142 . A rotation drive mechanism is provided on the edge of the RTP main valve, and a switch gate mechanism is provided on the RTP main valve.

[0090] The main valve body 141 is rotatably mounted on one of the side walls of the preparation chamber. The rotational drive mechanism includes a first rotational drive cylinder 143 and a second rotational drive cylinder 144, which extend and retract in parallel and opposite directions. These two rotational drive cylinders, positioned on either side of the main valve body 141, can drive the main valve body 141 to rotate. A main valve plate 142 seals the main valve body 141, thereby sealing the preparation chamber and preventing external contamination.

[0091] The door-opening mechanism is used to drive the main valve plate 142 to rotate, thereby opening or closing the RTP valve. Specifically, the door-opening mechanism includes a drive motor 145, a connecting rod assembly 146, a latch shaft 147, and a hinged leaf 148. The drive motor 145 is connected to the hinged leaf 148 via a gear. The drive motor 145 is also connected to the connecting rod assembly 146 via another gear. One side of the hinged leaf 148 is fixedly connected to the main valve plate 142. The drive motor 145 can drive the hinged leaf 148 to swing, thereby swinging the main valve plate 142 to open or close the main valve body 141.

[0092] A support block 149 is provided on the other side of the hinge movable page 148, and a latch seat 150 corresponding to the latch shaft 147 is provided on the main valve body 141. A through hole is provided on the support block 149, and the latch shaft 147 passes through the support block 149. The end thereof can be inserted into the latch seat 150 to lock the main valve plate 142 on the main valve body 141. The other end of the latch shaft 147 is hinged to the connecting rod assembly 146, and the driving motor 145 can control the movement of the latch shaft 147 through the connecting rod assembly 146 to unlock it.

[0093] A main valve disc is provided on the main valve plate 142, and the RTP auxiliary valve 151 is movably installed on the incubator 200. The auxiliary valve disc is provided on the RTP auxiliary valve 151. When the RTP main valve and the RTP auxiliary valve 151 are docked, the main valve plate 142 is rotated to a set angle, and the main valve disc and the auxiliary valve disc can be buckled together, thereby fixing the main valve plate 142 and the RTP auxiliary valve 151 together.

[0094] To improve the degree of automation, as shown in Figures 2, 3, 4, and 5, the cell preparation apparatus 100 further includes a control unit 161. A first in-position signal sensor 162, a second in-position signal sensor 163, a third in-position signal sensor 164, and a fourth in-position signal sensor 165 are also provided on the sidewalls of the preparation chamber. When the third in-position signal sensor 164 receives a signal, the control unit 161 obtains information indicating that the incubator 200 has been docked. At this point, the control unit 161 outputs a control signal to cause the first and second rotary drive cylinders 143, 144 to drive the main valve body 141 to rotate to a set angle.

[0095] When the first in-position signal sensor 162 loses its signal and the second in-position signal sensor 163 receives a signal, the first and second rotary drive cylinders 143 and 144 cease driving, allowing the main valve disc of the main valve plate 142 and the sub-valve disc of the RTP sub-valve 151 to engage, thereby securing the main valve plate 142 and the RTP sub-valve 151 together. At this point, the control unit 161 determines that the drive motor 145 is in operation, establishing interlocking feedback control. The drive motor 145 controls the movement of the latch shaft 147 via the connecting rod assembly 146, disengaging the end of the latch shaft 147 from the latch seat 150 to unlock the door. At this point, the drive motor 145 continues to rotate, swinging the hinge 148 so that the main valve plate 142 and the RTP sub-valve 151 swing, thereby opening the preparation chamber and incubator 200 of the preparation module.

[0096] When it is necessary to close the main valve body 141, the control unit 161 outputs a control signal to make the drive motor 145 rotate in the opposite direction, and the main valve plate 142 swings through the swing of the hinge movable leaf 148, thereby closing the main valve body 141. As the drive motor 145 continues to rotate, the end of the latch shaft 147 is inserted into the pin seat 150 for locking. When locked in place, the fourth in-place signal sensor 165 receives a signal to determine that the main valve body 141 has been locked.

[0097] Then, the control unit 161 outputs a control signal to make the first rotary drive cylinder 143 and the second rotary drive cylinder 144 drive the main valve body 141 to rotate in the opposite direction until the first in-position signal sensor 162 receives a signal and the second in-position signal sensor 163 loses a signal. At this time, it indicates that the main valve body 141 has rotated into position, the main valve disc and the auxiliary valve disc are no longer engaged, and the main valve plate 142 and the RTP auxiliary valve 151 are no longer fixed together. At this time, the preparation chamber of the preparation module and the incubator 200 are both sealed, and the two can be separated. After separation, the power socket 166 of the cell preparation device 100 no longer supplies power to the incubator 200.

[0098] In one embodiment, as shown in FIG2 , the cell preparation device 100 further includes a sterilization device for generating sterilization gas, and the output port of the sterilization device is connected to the preparation chamber for introducing sterilization gas into the preparation chamber for sterilization.

[0099] Specifically, in this embodiment, the sterilization device includes a liquid storage bottle 171, a delivery pump 172, a delivery pipeline 173 and a gas generating device 174. The sterilization gas is hydrogen peroxide gas. The liquid storage bottle 171 stores hydrogen peroxide liquid. The delivery pipeline 173 connects the liquid storage bottle 171 with the gas generating device 174. The delivery pump 172 delivers the hydrogen peroxide liquid to the gas generating device 174, and the gas generating device 174 converts the hydrogen peroxide liquid into hydrogen peroxide gas.

[0100] The cell preparation apparatus 100 is also equipped with a circulation fan 175, and the incubator 200 is equipped with a turbulence fan 210. The circulation fan 175 and turbulence fan 210 work together to fully diffuse the hydrogen peroxide gas within the preparation chamber and the incubation chamber 110, thereby achieving sufficient sterilization. The cell preparation apparatus 100 is also equipped with an exhaust assembly 176 to displace the hydrogen peroxide gas and exhaust it after sterilization. In other embodiments, other sterilizing gases can also be used for sterilization.

[0101] In one embodiment, in order to prevent the external environment from contaminating the incubator 200, a sterilizing filter is provided in the incubator 200, and a suction pipe 223 connected to the sterilizing filter is connected to the outside of the incubator 200. The suction pipe 223 sterilizes the sterilizing filter by sucking the sterilizing gas in the incubator 200.

[0102] The incubator 200 is also connected to a blowing pipe 227 connected to the sterilizing filter. The blowing pipe 227 is used to blow air into the sterilizing filter to discharge the sterilizing gas in the sterilizing filter after sterilization. At the same time, the blown gas will enter the culture chamber connected to the sterilizing filter to accelerate the turbulence.

[0103] Specifically, in this embodiment, to ensure adequate filtration and sterilization, the sterilizing filter is equipped with two sterilizing filters: a first sterilizing filter 221 and a second sterilizing filter 222. The incubator 200 is provided with a quick-connect pipe interface, to which both the suction pipe 223 and the air insufflation pipe 227 are connected. The suction pipe 223 is equipped with a first, two-position, two-way solenoid valve 225 and a vacuum pump 226, while the air insufflation pipe 227 is equipped with a second, two-position, two-way battery valve and a compressed air flow control valve 229. In other embodiments, a different number of sterilizing filters may be provided, such as three or four.

[0104] During sterilization, first open the first two-position two-way solenoid valve 225 and close the second two-position two-way solenoid valve 228, and use the suction of the vacuum pump 226 to allow the sterilizing gas in the incubator 200 to pass through the first sterilizing filter 221 and the second sterilizing filter 222 and enter the suction pipeline 223, so that the sterilizing gas can fully sterilize the two sterilizing filters.

[0105] After sterilization is completed, the exhaust component 176 is opened to discharge the sterilization gas. At the same time, the first two-position two-way battery valve is closed, the second two-position two-way solenoid valve 228 is opened, and the compressed air flow regulating valve 229 is kept open. Compressed air is introduced into the blowing pipe 227 to blow the first sterilizing filter 221 and the second sterilizing filter 222, so that the residual sterilization gas is separated from the two sterilizing filters and is replaced and discharged along with other sterilization gases.

[0106] In one embodiment, as shown in Figure 1, the sterile culture system of this embodiment also includes a transfer cart 400, which is used to transfer the incubator 200, for example, to transfer the incubator 200 from the cell culture device 300 to the cell preparation device 100, or to transfer the incubator 200 from the cell preparation device 100 to the incubator 200.

[0107] In one embodiment, the cell culture device 300 in this embodiment has a plurality of accommodating spaces arranged in an array. For example, the cell culture device 300 can be a honeycomb cell culture device 300, which can accommodate a plurality of culture boxes 200 at the same time for simultaneous culture. A control unit is provided inside the cell culture device 300 to centrally control the culture process of the plurality of culture boxes 200.

[0108] In one embodiment, the sterile culture system in this embodiment can implement the sterile culture method in any embodiment of the first aspect described above.

[0109] Before use, the incubator 200 is docked with the cell preparation apparatus 100 using a transfer cart 400. The preparation chamber and incubator 200 are then sterilized using a sterilizer. After sterilization, the incubator 200 and cell preparation apparatus 100 are separated. Cells are prepared within the cell preparation apparatus 100. After cell preparation is complete, the incubator 200 is re-docking with the cell preparation apparatus 100 and the cells are transferred to the incubator 200. The incubator 200 is then separated from the cell preparation apparatus 100, and the transfer cart 400 transfers the incubator 200 to the cell culture apparatus 300 for sterile culture. The entire cell transfer process is completed in under three minutes, is relatively simple, and effectively improves cell preparation and culture efficiency.

[0110] When periodic observation and replacement of culture medium are required during the culture process, the incubator 200 is transferred from the cell culture device 300 to the cell preparation device 100 again by the transfer cart 400, and the previous docking operation process is repeated to complete the docking. After observation and replacement of the culture medium, the incubator 200 is separated from the cell preparation device 100 again, and the transfer cart 400 is used to transfer the incubator 200 from the cell preparation device 100 to the cell culture device 300 to continue the culture, and this process is repeated until the culture is completed.

[0111] The entire culture process has low contamination risk and low personnel input costs. The transfer process has minimal impact on cell growth, avoiding transfer sterilization that kills the cultured cells. The culture process has low contamination risk and greatly improves cell preparation and culture efficiency.

[0112] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A sterile culture method, characterized in that: include: Providing a cell preparation device and a culture box; Sealing and docking the incubator with the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device are in communication with each other; Sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator, and after sterilization, sealing and separating the incubator from the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device remain sealed when separated; preparing cells in a preparation chamber of the cell preparation device; Sealing and docking the incubator with the cell preparation device so that the culture chamber of the incubator and the preparation chamber of the cell preparation device are in communication with each other, and transferring cells from the preparation chamber of the cell preparation device to the culture chamber of the incubator; Separating the incubator from the cell preparation device in a sealed manner, so that the culture chamber of the incubator and the preparation chamber of the cell preparation device remain sealed when separated; A cell culture device is provided, and the cell culture environment in the culture box is controlled by the cell culture device to culture the cells in the culture box.

2. The aseptic culture method according to claim 1, characterized in that The cell preparation device and the incubator are provided with sealing docking structures that cooperate with each other; The sealing docking structure comprises an RTP main valve and an RTP auxiliary valve, and the RTP main valve and the RTP auxiliary valve form an RTP valve when they are buckled together; The process of sealingly docking the incubator with the cell preparation device comprises: rotating the RTP main valve so that the RTP main valve and the RTP auxiliary valve are buckled to form an RTP valve; opening the RTP valve to achieve sealing docking of the incubator with the cell preparation device; The process of sealing and separating the incubator from the cell preparation device includes: closing the RTP valve; rotating the RTP main valve to separate the RTP main valve and the RTP auxiliary valve to achieve the The incubator is separated from the seal of the cell preparation device.

3. The aseptic culture method according to claim 1, characterized in that The process of sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator includes: Passing sterilizing gas into the cell preparation device and / or the incubator, and allowing the sterilizing gas to diffuse and circulate in the incubator and the cell preparation device; After the sterilization is completed, the sterilization gas in the preparation chamber and the culture chamber is exhausted.

4. The aseptic culture method according to claim 3, characterized in that: While sterilizing the preparation chamber of the cell preparation device and the culture chamber of the culture box, the sterilizing filter of the culture box is sterilized by the sterilizing gas, and the sterilizing filter is connected to the culture chamber through a gas pipeline.

5. A sterile culture system, characterized in that: It includes a cell preparation device, an incubator and a cell culture device; The cell preparation device comprises a preparation chamber, and a first sealing valve is arranged on the side wall of the preparation chamber; The incubator has a culture chamber, and a second sealing valve is provided on the side wall of the culture chamber, and the second sealing valve is used to cooperate with the first sealing valve to achieve sealed docking or sealed separation between the incubator and the cell preparation device; The cell culture device is used to accommodate a culture box, supply power and gas to the culture box, and control the cell growth environment in the culture box.

6. The sterile culture system according to claim 5, characterized in that: One of the first sealing valve and the second sealing valve is an RTP main valve, and the other is an RTP auxiliary valve. The RTP main valve and the RTP auxiliary valve form an RTP valve when they are buckled together.

7. The sterile culture system according to claim 6, characterized in that: The edge of the RTP main valve is provided with a first cylinder and a second cylinder for rotating the RTP main valve, and the extension and contraction directions of the first cylinder and the second cylinder are set to be parallel and opposite; The RTP main valve or the RTP auxiliary valve is provided with a switch gate mechanism, and the switch gate mechanism is used to open or close the RTP valve.

8. The sterile culture system according to claim 5, characterized in that: The cell preparation device further comprises a sterilization device for generating sterilization gas, and an output port of the sterilization device is communicated with the preparation chamber.

9. The sterile culture system according to claim 8, characterized in that: The cell preparation device further comprises a circulation fan, and a turbulence fan is arranged in the culture box. The circulation fan and the turbulence fan cooperate to fully diffuse the sterilization gas in the preparation chamber and the culture chamber.

10. The sterile culture system according to claim 8, characterized in that: The cell preparation device also includes an exhaust component, which is used to replace and discharge the sterilization gas after the sterilization is completed.

11. The sterile culture system according to claim 8, characterized in that: Also included: a vacuum pump and suction piping; A sterilizing filter is provided in the incubator, and the sterilizing filter is connected to the culture chamber through a gas pipeline; The suction pipeline is connected to the sterilizing filter, and the vacuum pump is arranged on the suction pipeline to suck the sterilizing gas in the incubator to sterilize the sterilizing filter.

12. The sterile culture system according to claim 11, characterized in that: Also includes: The air blowing pipeline is connected to the sterilizing filter and is used to blow air into the sterilizing filter so as to discharge the sterilizing gas in the sterilizing filter after the sterilization is completed.

13. The sterile culture system according to claim 5, characterized in that: The cell culture device has a plurality of accommodating spaces arranged in an array, and the culture boxes placed in the accommodating spaces are centrally controlled by the control unit of the cell culture device.

14. The sterile culture system according to claim 5, characterized in that: Also included is a transfer trolley, which is used to transfer the incubator.

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