Sterile culture method and sterile culture system

Abstract

The invention relates to the technical field of biopharmaceutical equipment, 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, and a first sealing valve is arranged on the side wall of the preparation chamber; the incubator is provided with a culture chamber, a second sealing valve is arranged on the side wall of the culture chamber, and the second sealing valve is used for being matched with the first sealing valve, so that the incubator and the cell preparation device are in sealed butt joint or sealed separation; the cell culture device is used for accommodating the culture box, supplying power and gas to the culture box and controlling the cell growth environment in the culture box. As the cells always grow in the sterile environment, the pollution risk is low, frequent sterilization is not needed, the normal growth of the cells is prevented from being influenced by frequent sterilization, the culture time is saved, and the cell preparation and culture efficiency is high.

Description

Technical Field

[0001] The present invention relates to the technical field of biopharmaceutical equipment, and particularly to a sterile culture method and a sterile culture system. Background Art

[0002] CGT (cell and gene therapy) products, as cutting-edge technologies in the pharmaceutical field, have high added value and huge market potential. They are part of the drugs for treating diseases. Like conventional chemical drugs, they also need to ensure the sterility of the production process during production. Once contaminated during the production process, patients may lose the best treatment opportunity due to the inability to obtain the drug in time, and in severe cases, it may lead to the death of the patient, causing significant losses to CGT pharmaceutical enterprises.

[0003] In addition, the cultivation of CGT drugs requires strict time control. If good time control is not carried out during the preparation and cultivation processes, it will affect the cell growth status, and the cell culture conditions will directly affect the treatment effect of patients receiving cell therapy. CGT drugs are biologically active drugs, and a series of operations such as observation and medium replacement need to be carried out frequently during the cultivation process. The traditional method is to prepare the cell drug in the cell preparation device and then transfer it to a conventional incubator for cultivation. When operations such as observation and medium replacement are required during the cultivation process, it is transferred back to the cell preparation device for processing, and then transferred back to the incubator for continued cultivation, with continuous repeated operations.

[0004] During the process of reciprocating movement of the cell drug from the cell preparation device to the incubator, the cell culture container will pass through the B / C / D low-level environments of the laboratory, and the probability of cell contamination is extremely high; the personnel performing the transfer operation need to have very professional skills, and the situation of the operator will directly affect the cell contamination risk during the transfer process; the entire cultivation process of the conventional incubator is in a low-level environment, and the contamination risk during the cell cultivation process is very high. In addition, when frequently sterilizing the culture container containing cells, there is a risk that the sterilizing agent will penetrate the cell container and kill the cells. Therefore, the existing sterile culture method has problems of high contamination risk and low cell preparation and cultivation efficiency during the cell cultivation process. Summary of the Invention

[0005] 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 cultivation efficiency in the prior art during the process of culturing cells.

[0006] To solve the above technical problems, the present invention provides a sterile culture method.

[0007] The sterile culture method of the present invention includes:

[0008] Providing a cell preparation device and an incubator;

[0009] Seal and dock the incubator with the cell preparation device so that the culture chamber of the incubator communicates with the preparation chamber of the cell preparation device;

[0010] Sterilize the preparation chamber of the cell preparation device and the culture chamber of the incubator. After sterilization, 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 when separated;

[0011] Prepare cells in the preparation chamber of the cell preparation device;

[0012] Seal and dock the incubator with the cell preparation device so that the culture chamber of the incubator communicates with the preparation chamber of the cell preparation device, and transfer the cells from the preparation chamber of the cell preparation device to the culture chamber of the incubator;

[0013] 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 when separated;

[0014] Provide a cell culture device and control the cell culture environment in the incubator through the cell culture device to culture the cells in the incubator.

[0015] Further, the cell preparation device and the incubator are provided with mutually cooperating seal docking structures;

[0016] The seal docking structure includes an RTP main valve and an RTP sub - valve. When the RTP main valve and the RTP sub - valve are buckled together, they form an RTP valve;

[0017] The process of the seal docking of the incubator and the cell preparation device includes: by rotating the RTP main valve, the RTP main valve and the RTP sub - valve are buckled together to form an RTP valve; open the RTP valve to achieve the seal docking of the incubator and the cell preparation device;

[0018] The process of the seal separation of the incubator and the cell preparation device includes: closing the RTP valve; by rotating the RTP main valve, the separation of the RTP main valve and the RTP sub - valve is achieved to realize the seal separation of the incubator and the cell preparation device.

[0019] Further, the process of sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator includes:

[0020] 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;

[0021] After the sterilization is completed, the sterilization gas in the preparation chamber and the culture chamber is exhausted.

[0022] Furthermore, while sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator, the sterilizing gas is used to sterilize the sterilizing filter of the incubator, and the sterilizing filter is connected to the culture chamber through a gas pipeline.

[0023] The present invention also provides a sterile culture system, comprising a cell preparation device, a culture box and a cell culture device;

[0024] The cell preparation device comprises a preparation chamber, and a first sealing valve is arranged on the side wall of the preparation chamber;

[0025] 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;

[0026] 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.

[0027] Further, one of the first sealing valve and the second sealing valve is an RTP main valve, and the other is an RTP auxiliary valve, and the RTP main valve and the RTP auxiliary valve form an RTP valve when they are buckled together.

[0028] Further, 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 retraction directions of the first cylinder and the second cylinder are set to be parallel and opposite;

[0029] 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.

[0030] Furthermore, the cell preparation device also includes a sterilization device for generating sterilization gas, and an output port of the sterilization device is connected to the preparation chamber.

[0031] Furthermore, the cell preparation device also includes a circulating fan, and the incubator is provided with a turbulent fan. The circulating fan and the turbulent fan cooperate to fully diffuse the sterilizing gas in the preparation chamber and the culture chamber.

[0032] Further, the cell preparation device further includes an exhaust component, which is used to displace and exhaust the sterilization gas after the sterilization is completed.

[0033] Further, it also includes: a vacuum pump and a suction pipeline;

[0034] A sterilization filter is provided in the incubator, and the sterilization filter is communicated with the culture chamber through a gas pipeline;

[0035] The suction pipeline is connected to the sterilization filter, and the vacuum pump is arranged on the suction pipeline for sucking the sterilization gas in the incubator to sterilize the sterilization filter.

[0036] Further, it also includes: a blowing pipeline, which is connected to the sterilization filter and is used to blow the sterilization filter to discharge the sterilization gas in the sterilization filter after the sterilization is completed.

[0037] Further, the cell culture device has a plurality of accommodation spaces arranged in an array, and the incubators placed in the accommodation spaces are all centrally controlled by the control unit of the cell culture device.

[0038] Further, it also includes a transfer cart, which is used to transfer the incubator.

[0039] Compared with the prior art, the present invention has at least the following beneficial effects:

[0040] Before use, first seal and dock the incubator and the cell preparation device so that the gas environment of the culture chamber and the gas environment of the preparation chamber are communicated, and then sterilize the two. After the sterilization is completed, seal and separate the two. Then prepare cells in the preparation chamber. When the cell preparation is completed, cooperate the second sealing valve of the incubator with the first sealing valve of the cell preparation device, so that the incubator and the cell preparation device are sealed and docked, and then the gas environment of the culture chamber and the gas environment of the preparation chamber are communicated. In this sealed and sterile environment, transfer the cells to the incubator, and then seal and separate the incubator and the cell preparation device, and transfer the incubator to the cell culture device to culture the cells.

[0041] Since the cell preparation device and the incubator have been sterilized before preparing cells, the prepared cells grow in a sterile environment. Also, because the incubator and the cell preparation device are sealed and docked before transferring the cells 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 lower risk of contamination. Therefore, it is not necessary to sterilize frequently, which not only avoids affecting the normal growth of cells due to frequent sterilization but also saves the culture time. Therefore, the cell preparation and culture efficiency are relatively high. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 Schematic structural diagram of an embodiment of the aseptic culture system of the present invention;

[0043] Figure 2 is Figure 1 Schematic structural diagram when the cell preparation device and the incubator in [[ ]] are sterilized together after docking;

[0044] Figure 3 is Figure 1 Schematic structural diagram after hiding part of the structure when the cell preparation device and the incubator in [[ ]] are docked;

[0045] Figure 4 is Figure 1 Schematic structural diagram when part of the structure of the cell preparation device in [[ ]] is hidden to show the RTP main valve;

[0046] Figure 5 is Figure 4 Local enlarged view of part A in [[ ]];

[0047] Reference numerals:

[0048] 100, cell preparation device; 110, culture chamber; 120, support frame; 130, limiting member; 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 page; 149, support block; 150, plug seat; 151, RTP sub-valve; 161, control unit; 162, first in-place signal sensor; 163, second in-place signal sensor; 164, third in-place signal sensor; 165, fourth in-place signal sensor; 166, power supply socket; 171, liquid storage bottle; 172, delivery pump; 173, delivery pipeline; 174, gas generating device; 175, circulation fan; 176, exhaust assembly;

[0049] 200, incubator; 210, turbulence fan; 221, first sterilization filter; 222, second sterilization filter; 223, suction pipeline; 225, first two-position two-way solenoid valve; 226, vacuum pump; 227, blow pipeline; 228, second two-position two-way solenoid valve; 229, compressed air flow regulating valve;

[0050] 300, cell culture device;

[0051] 400, transfer trolley. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0052] The following will describe a sterile culture method and a sterile culture system of the present invention in combination with schematic diagrams, in which the preferred embodiments of the present invention are shown. It should be understood that those skilled in the art can modify the present invention described herein while still achieving the advantageous effects of the present invention. Therefore, the following description should be understood as a broad guidance for those skilled in the art and not as a limitation on the present invention.

[0053] The serial numbers assigned to the components herein, such as "first", "second", etc., are only used to distinguish the described objects and do not have any sequential or technical meanings. And the "connection" and "coupling" mentioned in this application, unless otherwise specified, both include direct and indirect connection (coupling). In the description of the present invention, it should be understood that the orientation or positional relationship indicated by the terms "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc. is based on the orientation or positional relationship shown in the drawings, and is 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 a limitation on the present invention.

[0054] In the present invention, unless otherwise clearly specified and limited, the first feature being "on" or "under" the second feature may be that the first and second features are in direct contact, or the first and second features are indirectly in contact through an intermediate medium. Moreover, the first feature being "above", "over" and "on" the second feature may be that the first feature is directly above or obliquely above the second feature, or simply means that the first feature has a higher horizontal height than the second feature. The first feature being "under", "beneath" and "under" the second feature may be that the first feature is directly below or obliquely below the second feature, or simply means that the first feature has a lower horizontal height than the second feature.

[0055] In the following paragraphs, the present invention will be described more specifically by way of example with reference to the drawings. According to the following description and the claims, the advantages and features of the present invention will be clearer. It should be noted that the drawings are all in a very simplified form and use non-precise scales, and are only used to conveniently and clearly assist in explaining the purpose of the embodiments of the present invention.

[0056] The following will introduce the sterile culture method in the first aspect embodiment of the present invention in combination with the specification.

[0057] In one of the embodiments, the sterile culture method of this embodiment includes:

[0058] S100: Provide a cell preparation device and an incubator;

[0059] S200: Hermetically dock the incubator with the cell preparation device so that the culture chamber of the incubator communicates with the preparation chamber of the cell preparation device;

[0060] S300: Sterilize the preparation chamber of the cell preparation device and the culture chamber of the incubator. After sterilization, hermetically 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 when separated;

[0061] S400: Prepare cells in the preparation chamber of the cell preparation device;

[0062] S500: Hermetically dock the incubator with the cell preparation device so that the culture chamber of the incubator communicates with the preparation chamber of the cell preparation device, and transfer the cells from the preparation chamber of the cell preparation device to the culture chamber of the incubator;

[0063] S600: Hermetically 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 when separated;

[0064] S700: Provide a cell culture device and control the cell culture environment in the incubator through the cell culture device to culture the cells in the incubator.

[0065] In step S300, hydrogen peroxide gas can be used to sterilize the cell preparation device and the incubator. In other embodiments, other types of sterilization gases can also be used, and other sterilization methods can also be adopted for sterilization, such as using ultraviolet sterilization.

[0066] In step S500, the docking process needs to ensure sealing to avoid external environmental pollution. After docking, the gas environments in the cell preparation device and the incubator are connected. Since both are sterile environments, the transfer process is also completed in a sterile process, and the pollution risk is relatively low.

[0067] Since the cell preparation device and the incubator have been sterilized before cell preparation, the prepared cells grow in a sterile environment. Also, because the incubator and the cell preparation device are hermetically 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 relatively low pollution risk. Therefore, frequent sterilization is not required, which not only avoids affecting the normal growth of cells due to frequent sterilization but also saves the culture time. Therefore, the cell preparation and culture efficiency are relatively high.

[0068] In one embodiment, the cell preparation device and the incubator are provided with a sealing and docking structure that cooperates with each other. The sealing and docking structure includes an RTP (Rapid Transfer Ports) main valve and an RTP sub-valve. When the RTP main valve and the RTP sub-valve are buckled together, they form an RTP valve. After the RTP main valve and the RTP sub-valve are buckled together, the outer surface of the RTP main valve and the outer surface of the RTP sub-valve are sealed in the RTP valve. Herein, 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 sub-valve. Opening the RTP valve formed by buckling the RTP main valve and the RTP sub-valve can connect two sterile chambers.

[0069] The process of using the sealing and docking structure to achieve the sealed docking of the incubator and the cell preparation device includes: rotating the RTP main valve so that the RTP main valve and the RTP sub-valve are buckled together to form an RTP valve; opening the RTP valve to achieve the sealed docking of the incubator and the cell preparation device.

[0070] The process of using the sealing and docking structure to achieve the sealed separation of the incubator and the cell preparation device includes: closing the RTP valve; rotating the RTP main valve so that the RTP main valve and the RTP sub-valve are separated to achieve the sealed separation of the incubator and the cell preparation device.

[0071] By using the RTP main valve and the RTP sub-valve, the sealed docking or sealed separation of the cell preparation device and the incubator can be automatically achieved. Transferring cells is relatively convenient, without the need for professional skills. General laboratory assistants can complete the entire transfer process through simple operations, with a low investment in labor costs and no pollution during the transfer process. Preferably, the RTP main valve is provided on the cell preparation device, and the RTP sub-valve is provided on the incubator. Of course, in other embodiments, the RTP sub-valve can also be provided on the cell preparation device, and the RTP main valve is provided on the incubator.

[0072] In one embodiment, in order to sterilize the cell preparation device and the incubator, before preparing cells, the incubator and the cell preparation device are sealed and docked, and a sterilization gas is introduced into the cell preparation device and / or the incubator to allow the sterilization gas to diffuse and circulate in the incubator and the cell preparation device.

[0073] The process of sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator includes:

[0074] Introducing a sterilization gas into the cell preparation device and / or the incubator, and allowing the sterilization gas to diffuse and circulate in the incubator and the cell preparation device;

[0075] After the sterilization is completed, the sterilization gas in the preparation chamber and the culture chamber is discharged.

[0076] Among them, the sterilization gas is preferably hydrogen peroxide gas, and the hydrogen peroxide gas can be generated from hydrogen peroxide liquid. The sterilization gas can be introduced into the cell preparation device, and then diffused and circulated by means of a turbulence fan or other turbulence components. Of course, the sterilization gas can also be first introduced into the incubator, or the sterilization gas can be introduced into the cell preparation device and the incubator at the same time. In addition, other types of sterilization gas can be selected according to needs.

[0077] In order to avoid environmental pollution of the incubator by the outside world, a sterilization filter is provided outside the culture chamber, and the sterilization filter is communicated with the culture chamber through a gas pipeline. In one embodiment, while sterilizing the preparation chamber of the cell preparation device and the culture chamber of the incubator, the sterilization filter of the incubator is sterilized by using the sterilization gas.

[0078] The following combines the specification appendix Figures 1 to 5 to introduce the aseptic culture system in the second aspect embodiment of the present invention.

[0079] As Figure 1 and Figure 2 shown, in one embodiment, the aseptic culture system of this embodiment includes a cell preparation device 100, an incubator 200, and a cell culture device 300.

[0080] The cell preparation device 100 includes a preparation chamber, and a first sealing valve is provided on the side wall of the preparation chamber.

[0081] 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 between the incubator 200 and the cell preparation device 100.

[0082] The cell culture device 300 is used to accommodate the incubator 200, supply power and gas to the incubator, and control the cell growth environment in the incubator.

[0083] Before use, first seal and dock the incubator 200 and the cell preparation device 100 so that the gas environment in the culture chamber 110 is connected to the gas environment in the preparation chamber, and then sterilize the two. After sterilization, seal and separate them. Then prepare cells in the preparation chamber. When the cell preparation is completed, cooperate the second sealing valve of the incubator 200 with the first sealing valve of the cell preparation device 100 to seal and dock the incubator 200 and the cell preparation device 100, so that the gas environment in the culture chamber 110 is connected to the gas environment in the preparation chamber. In this sealed and sterile environment, transfer the cells to the incubator 200, and then seal and separate the incubator 200 and the cell preparation device 100, and transfer the incubator 200 into the cell culture device 300 to culture the cells.

[0084] Since the cell preparation device 100 and the incubator 200 have been sterilized before cell preparation, the prepared cells grow in a sterile environment. Also, because the incubator 200 and the cell preparation device 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, with a lower risk of contamination. Therefore, frequent sterilization is not required, which not only avoids affecting the normal growth of cells due to frequent sterilization but also saves the culture time. Thus, the cell preparation and culture efficiency are relatively high.

[0085] Specifically, the cell preparation device 100 further includes a support frame 120 for supporting the incubator 200. A limiting member 130 is provided on the support frame 120, and the limiting member 130 is used to limit the position of the incubator 200 to firmly fix the incubator 200 on the support frame 120.

[0086] To achieve aseptic transfer, an aseptic operation window is provided on the side wall of the preparation chamber, and aseptic operation gloves, such as rubber gloves, are provided at the aseptic operation window. The aseptic operation gloves are located inside the preparation chamber and are sealed with the aseptic operation window. The operator only needs to insert both hands into the aseptic operation gloves to perform operations without opening the preparation chamber, such as transferring the cells in the preparation chamber to the docked incubator 200, or transferring the cells in the docked incubator 200 to the preparation chamber.

[0087] In one embodiment, such as Figure 3 、 Figure 4 and Figure 5As shown, 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 sub-valve 151. Preferably, the first sealing valve is the RTP main valve, and the second sealing valve is the RTP sub-valve 151, that is, the RTP main valve is provided in the cell preparation device 100, and the RTP sub-valve 151 is provided in the incubator 200.

[0088] After the RTP main valve and the RTP sub-valve 151 are buckled together, the outer surface of the RTP main valve and the outer surface of the RTP sub-valve 151 are sealed in the RTP valve. Among them, 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 sub-valve 151. Opening the RTP valve formed by buckling the RTP main valve and the RTP sub-valve 151 can connect two sterile chambers.

[0089] The process of realizing the sealed docking of the incubator and the cell preparation device by using the sealed docking structure includes: rotating the RTP main valve so that the buckling of the RTP main valve and the RTP sub-valve 151 forms an RTP valve; opening the RTP valve to realize the sealed docking of the incubator and the cell preparation device.

[0090] The process of realizing the sealed separation of the incubator and the cell preparation device by using the sealed docking structure includes: closing the RTP valve; rotating the RTP main valve so that the RTP main valve and the RTP sub-valve 151 are separated to realize the sealed separation of the incubator and the cell preparation device.

[0091] By using the RTP main valve and the RTP sub-valve 151, the sealed docking or sealed separation of the cell preparation device 100 and the incubator 200 can be automatically realized, which is more convenient for transferring cells. No professional skills are required, and general laboratory assistants can complete the entire transfer process through simple operations, with a relatively low investment in labor costs. Of course, in other embodiments, the RTP sub-valve 151 can also be used as the first sealing valve, and the RTP main valve can be used as the second sealing valve.

[0092] Specifically, the RTP main valve includes a main valve body 141 and a main valve plate 142. A rotation driving mechanism is provided on the edge of the RTP main valve, and a door opening and closing mechanism is provided on the RTP main valve.

[0093] The main valve body 141 is rotatably disposed on one of the side walls of the preparation chamber, and the rotation drive mechanism includes a first rotation drive cylinder 143 and a second rotation drive cylinder 144 whose extension and contraction directions are parallel and opposite to each other. The two rotation drive cylinders are disposed on both sides of the main valve body 141 and can drive the main valve body 141 to rotate. The main valve plate 142 can block the main valve body 141, thereby sealing the preparation chamber to prevent external contamination.

[0094] The door opening and closing mechanism is used to drive the main valve plate 142 to rotate so as to open or close the RTP valve. Specifically, the door opening and closing mechanism includes a driving motor 145, a connecting rod assembly 146, a latch shaft 147 and a hinge movable leaf 148. The driving motor 145 is connected to the hinge movable leaf 148 through a gear, and the driving motor 145 is also connected to the connecting rod assembly 146 through other gears. One side of the hinge movable leaf 148 is fixedly connected to the main valve plate 142. The driving motor 145 can drive the hinge movable leaf 148 to swing, thereby swinging the main valve plate 142 to open or block the main valve body 141.

[0095] A support block 149 is provided on the other side of the hinge movable leaf 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 for unlocking.

[0096] A main valve disc is arranged on the main valve plate 142, and the RTP auxiliary valve 151 is movably installed on the incubator 200. An auxiliary valve disc is arranged 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.

[0097] In order to improve the degree of automation, such as Figure 2 , Figure 3 , Figure 4 and Figure 5As shown, the cell preparation device 100 further includes a control unit 161. On the side wall of the preparation chamber, there are also provided a first in-place signal sensor 162, a second in-place signal sensor 163, a third in-place signal sensor 164, and a fourth in-place signal sensor 165. Among them, when the third in-place signal sensor 164 receives a signal, the control unit 161 obtains information indicating that the incubator 200 has been docked in place. At this time, the control unit 161 outputs a control signal to cause the first rotary drive cylinder 143 and the second rotary drive cylinder 144 to drive the main valve body 141 to rotate by a set angle.

[0098] When the first in-place signal sensor 162 loses the signal and the second in-place signal sensor 163 obtains the signal, the first rotary drive cylinder 143 and the second rotary drive cylinder 144 stop driving, and the main valve piece of the main valve plate 142 and the sub-valve piece of the RTP sub-valve 151 can be buckled together, thereby fixing the main valve plate 142 and the RTP sub-valve 151 together. At this time, the control unit 161 determines that the drive motor 145 is in an operating state, forms an interlock feedback control, and the drive motor 145 controls the movement of the latch shaft 147 through the link assembly 146. The end of the latch shaft 147 disengages from the latch seat 150 to perform unlocking. At this time, the drive motor 145 continues to rotate, and swings the main valve plate 142 to drive the RTP sub-valve 151 to swing through the hinge movable page 148, so as to open the preparation chamber of the preparation module and the incubator 200.

[0099] When it is necessary to close the main valve body 141, the control unit 161 outputs a control signal to cause the drive motor 145 to rotate in the reverse direction, swings the main valve plate 142 through the hinge movable page 148, and then closes the main valve body 141. As the drive motor 145 continues to rotate, the end of the latch shaft 147 is inserted into the latch seat 150 to perform locking. When the locking is in place, the fourth in-place signal sensor 165 receives a signal to determine that the main valve body 141 has been locked.

[0100] Then, the control unit 161 outputs a control signal to cause the first rotary drive cylinder 143 and the second rotary drive cylinder 144 to drive the main valve body 141 to rotate in the reverse direction until the first in-place signal sensor 162 obtains the signal and the second in-place signal sensor 163 loses the signal. At this time, it indicates that the main valve body 141 has rotated in place, the main valve piece and the sub-valve piece no longer buckle together, and the main valve plate 142 and the RTP sub-valve 151 are no longer fixed together. At this time, both the preparation chamber of the preparation module and the incubator 200 are sealed and can be separated. After separation, the power supply socket 166 of the cell preparation device 100 no longer supplies power to the incubator 200.

[0101] In one embodiment, as Figure 2As shown, the cell preparation device 100 further includes a sterilization device for generating sterilization gas. The output port of the sterilization device is communicated with the preparation chamber, and is used to introduce sterilization gas into the preparation chamber for sterilization.

[0102] Specifically, in this embodiment, the sterilization device includes a liquid storage bottle 171, a delivery pump 172, a delivery pipeline 173, and a gas generation 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 and the gas generation device 174. The delivery pump 172 delivers the hydrogen peroxide liquid to the gas generation device 174, and the gas generation device 174 converts the hydrogen peroxide liquid into hydrogen peroxide gas.

[0103] A circulation fan 175 is further provided in the cell preparation device 100, and a turbulence fan 210 is provided in the incubator 200. The circulation fan 175 and the turbulence fan 210 cooperate to fully diffuse the hydrogen peroxide gas in the preparation chamber and the culture chamber 110 for sufficient sterilization. An exhaust assembly 176 is further provided in the cell preparation device 100 to displace and exhaust the hydrogen peroxide gas after sterilization. In other embodiments, other sterilization gases can also be used for sterilization.

[0104] In one embodiment, in order to avoid environmental pollution of the incubator 200 by the outside world, a sterilization filter is provided in the incubator 200. An aspiration pipeline 223 communicated with the sterilization filter is connected outside the incubator 200. The aspiration pipeline 223 aspirates the sterilization gas in the incubator 200 to sterilize the sterilization filter.

[0105] A blow pipeline 227 communicated with the sterilization filter is further connected outside the incubator 200. The blow pipeline 227 is used to blow the sterilization filter to discharge the sterilization gas in the sterilization filter after sterilization. At the same time, the blown gas will enter the culture chamber communicated with the sterilization filter to accelerate turbulence.

[0106] Specifically, in this embodiment, in order to fully filter and sterilize, two sterilization filters are provided in the sterilization filter, namely a first sterilization filter 221 and a second sterilization filter 222. A pipeline quick-connect interface is provided on the incubator 200, and the aspiration pipeline 223 and the blow pipeline 227 are simultaneously connected to the pipeline quick-connect interface. A first two-position two-way solenoid valve 225 and a vacuum pump 226 are provided on the aspiration pipeline 223, and a second two-position two-way solenoid valve and a compressed air flow regulating valve 229 are provided on the blow pipeline 227. In other embodiments, different numbers of sterilization filters can also be provided, such as three or four.

[0107] During sterilization, first open the first two-position two-way solenoid valve 225 and close the second two-position two-way solenoid valve 228. By the suction of the vacuum pump 226, the sterilization gas in the incubator 200 passes through the first sterilization filter 221 and the second sterilization filter 222 and enters the suction pipeline 223, so that the sterilization gas can fully sterilize the two sterilization filters.

[0108] After sterilization, turn on the exhaust assembly 176 to exhaust the sterilization gas. At the same time, close the first two-position two-way solenoid valve, open the second two-position two-way solenoid valve 228, and keep the compressed air flow regulating valve 229 open. Introduce compressed air into the blow pipeline 227 to blow the first sterilization filter 221 and the second sterilization filter 222, so that the residual sterilization gas detaches from the two sterilization filters and is displaced and exhausted along with other sterilization gases.

[0109] In one embodiment, as Figure 1 shown, the aseptic culture system of this embodiment further includes a transfer cart 400, and the transfer cart 400 is used to transfer the incubator 200. For example, transfer the incubator 200 from the cell culture device 300 to the cell preparation device 100, or transfer the incubator 200 from the cell preparation device 100 to the incubator 200.

[0110] In one embodiment, the cell culture device 300 in this embodiment has a plurality of accommodation spaces arranged in an array. For example, the cell culture device 300 can be a honeycomb cell culture device 300, and multiple incubators 200 can be accommodated inside 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 multiple incubators 200.

[0111] In one embodiment, the aseptic culture system in this embodiment can implement the aseptic culture method in any of the above first aspects.

[0112] Before use, first use the transfer cart 400 to dock the incubator 200 with the cell preparation device 100, and then use the sterilization device to sterilize the preparation chamber and the incubator 200. After sterilization, separate the incubator 200 from the cell preparation device 100. Prepare cells in the cell preparation device 100. After cell preparation is completed, dock the incubator 200 with the cell preparation device 100 again, transfer the cells into the incubator 200, and then separate the incubator 200 from the cell preparation device 100. The transfer cart 400 transfers the incubator 200 into the cell culture device 300 for aseptic culture. The entire cell transfer process can be completed within 3 minutes, with relatively simple operation, and can effectively improve the efficiency of cell preparation and culture.

[0113] When it is necessary to periodically observe and replace the culture medium during the culture process, the incubator 200 is transferred from the cell culture device 300 to the cell preparation device 100 again through the transfer cart 400. The previous docking operation process is repeated to complete the docking. After observing and replacing the culture medium, the incubator 200 is separated from the cell preparation device 100 again. The incubator 200 is transferred from the cell preparation device 100 to the cell culture device 300 by using the transfer cart 400, and the culture is continued. This is repeated until the culture is completed.

[0114] The pollution risk during the whole culture process is small, the personnel input cost is low, the impact of the transfer process on cell growth is extremely low, the transfer sterilization is avoided from killing the cultured cells, the pollution risk during the culture process is small, and at the same time, the cell preparation and culture efficiency are greatly improved.

[0115] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these modifications and variations.

Claims

1. Aseptic culture method, It is 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, It is 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 comprises: 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.

3. The aseptic culture method according to claim 1, It is 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, It is 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, It is 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. 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 the incubator, supply power and gas to the incubator, and control the cell growth environment in the incubator.

6. The sterile culture system according to claim 5, wherein, One of the first sealing valve and the second sealing valve is an RTP main valve, and the other is an RTP sub-valve. When the RTP main valve and the RTP sub-valve are buckled together, they form an RTP valve.

7. The sterile culture system according to claim 6, wherein, A first cylinder and a second cylinder for rotating the RTP main valve are provided at the edge of the RTP main valve. The telescopic directions of the first cylinder and the second cylinder are set to be parallel and opposite; A switch door mechanism is provided on the RTP main valve or the RTP sub-valve. The switch door mechanism is used to open or close the RTP valve.

8. The sterile culture system according to claim 5, wherein, The cell preparation device further includes a sterilization device for generating sterilizing gas. The output port of the sterilization device is communicated with the preparation chamber.

9. The sterile culture system according to claim 8, wherein, The cell preparation device further includes a circulation fan, and a turbulence fan is provided in the incubator. The circulation fan and the turbulence fan cooperate to fully diffuse the sterilizing gas in the preparation chamber and the culture chamber.

10. The sterile culture system according to claim 8, wherein, The cell preparation device further includes an exhaust component, and the exhaust component is used to replace and exhaust the sterilizing gas after sterilization.

11. The sterile culture system according to claim 8, wherein, It further includes: a vacuum pump and a suction pipeline; A sterilization filter is provided in the incubator, and the sterilization filter is communicated with the culture chamber through a gas pipeline; The suction pipeline is connected to the sterilization filter, and the vacuum pump is arranged on the suction pipeline for sucking the sterilizing gas in the incubator to sterilize the sterilization filter.

12. The sterile culture system according to claim 11, wherein, It further includes: A blow pipeline, the blow pipeline is connected to the sterilization filter, and is used to blow the sterilization filter to discharge the sterilizing gas in the sterilization filter after sterilization.

13. The sterile culture system according to claim 5, wherein, The cell culture device has a plurality of accommodation spaces arranged in an array, and the incubators placed in the accommodation spaces are all centrally controlled by the control unit of the cell culture device.

14. The sterile culture system according to claim 5, wherein, It further includes a transfer cart, and the transfer cart is used to transfer the incubator.

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