WO2017090773A1 - Système de culture cellulaire et procédé de stérilisation - Google Patents

Système de culture cellulaire et procédé de stérilisation Download PDF

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Publication number
WO2017090773A1
WO2017090773A1 PCT/JP2016/085234 JP2016085234W WO2017090773A1 WO 2017090773 A1 WO2017090773 A1 WO 2017090773A1 JP 2016085234 W JP2016085234 W JP 2016085234W WO 2017090773 A1 WO2017090773 A1 WO 2017090773A1
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WIPO (PCT)
Prior art keywords
gas
line
module
sterilization
main
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PCT/JP2016/085234
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English (en)
Japanese (ja)
Inventor
久 五味
敏光 冨士
成則 尾▲崎▼
高橋　勉
明威 田村
竹内　晴紀
小林　直樹
佳雅 須田
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東京エレクトロン株式会社
シンフォニアテクノロジー株式会社
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Publication of WO2017090773A1 publication Critical patent/WO2017090773A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/16—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using chemical substances
    • A61L2/20—Gaseous substances, e.g. vapours
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/24—Apparatus using programmed or automatic operation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/12—Apparatus for enzymology or microbiology with sterilisation, filtration or dialysis means

Definitions

  • the present invention relates to a cell culture system including a plurality of operation modules that automatically perform operations necessary for cell culture, and a transfer module that automatically transfers a culture container such as a medium plate between the operation modules.
  • the present invention relates to a technique for sterilizing the inside.
  • the cell culture system In addition to an incubator module that is maintained in an environment suitable for cell culture in order to perform cell culture, the cell culture system includes a work module for performing various operations such as culture preparation, post-treatment, inspection, and observation. included. The cell culture system also includes a transfer module that transfers the culture container between the work modules.
  • An object of the present invention is to provide a technique capable of efficiently sterilizing each module of a cell culture system formed by combining a plurality of modules and efficiently purging sterilization gas after sterilization treatment. It is said.
  • a transport module having a transport machine for transporting a culture container therein, and connected to the transport module, the culture container or the contents of the culture container are related to culture.
  • a plurality of work modules including at least a culture module for culturing cells and a test module for inspecting a state of cells cultured in the culture module.
  • a plurality of branch supply lines each branching into a plurality of branch supply lines each supplying gas from the main gas line to the corresponding work module, and branching in parallel from the second region of the main gas line
  • a plurality of branch discharge lines each of which discharges gas from the corresponding work module to the main gas line, and a supply on / off valve provided in each branch supply line,
  • a cell culture system provided with a discharge on-off valve provided in each branch discharge line.
  • the sterilization method performed in the cell culture system selecting a sterilization target module to be sterilized from the plurality of operation modules, and for dividing the main gas line With the on-off valve closed, the supply on-off valve of the branch supply line and the discharge on-off valve of the branch discharge line corresponding to the module to be sterilized are opened, and then the first region side of the main gas line Thereby supplying the sterilizing gas to the first region of the main gas line, the branch supply line corresponding to the module to be sterilized, the inside of the module to be sterilized, the branch discharge line corresponding to the module to be sterilized, and Sterilizing the inside of the module to be sterilized by sequentially flowing the sterilizing gas through the second region of the main gas line And then supplying a purge gas to the first region side of the main gas line, whereby the first region of the main gas line, the branch supply line corresponding to the module to be sterilized, and the module to be sterilized
  • the main gas line can be used as the first region for gas supply and the second region for gas discharge by closing the dividing on-off valve.
  • the sterilization gas can be supplied to any sterilization target module from a common gas supply source by opening the supply on / off valve of the gas supply line and the discharge on / off valve of the gas return line corresponding to the sterilization target module. It is possible to sterilize any module to be sterilized efficiently.
  • the entire main gas line can be reliably purged by flowing the purge gas through the main gas line with the dividing on-off valve opened.
  • the cell culture system 1 includes a transfer module 2, a plate loader module 3 connected to the transfer module 2, and a plurality of work modules 4.
  • the transfer module 2 is formed by joining two transfer module parts 2A and a transfer module part 2B.
  • a transfer robot 82 (see FIG. 2) is provided in the internal space (transfer space) of the transfer module 2, and a culture plate (not shown) and a CI (chemical indicator) are provided between the plate loader module 3 and the work module 4. ) And BI (biological indicators) (both not shown).
  • the plate loader module 3 is configured as a load port (container loading / unloading section) that receives and dispenses a cassette (not illustrated) that accommodates a plurality of (for example, 16) culture plates (not illustrated) as a kind of culture container. Has been.
  • Each of the work modules 4 has a function specialized for executing various work (culture, pre-treatment and post-treatment of culture, post-culture inspection, etc.) performed for cell culture.
  • the work module 4 includes a plate loader module, an incubator module (culture module), an imaging module, a microscope module, a remover module, a preparation module, a liquid exchange module, a medium analysis module, and the like.
  • the incubator module is a module in which cell culture is performed, and includes a device for maintaining an environment (temperature, humidity, carbon dioxide concentration, etc.) suitable for cell culture.
  • the imaging module and the microscope module include devices for performing macroscopic and microscopic imaging of cultured cells.
  • the preparation module has a device for performing processing for culture preparation (ECM (Extracellular matrix) coating, PBS (Phosphate buffered saline) processing, etc.) on the culture plate.
  • the module has a device that removes defective cells based on imaging results of an imaging module, a microscope module, etc.
  • the liquid exchange module is used for seeding, periodic medium replacement, taking out cultured cells, etc.
  • the medium analysis module has a device for analyzing the medium, and the work module can be equipped with a device related to cell culture other than the above-described devices. .
  • the cell culture system 1 further includes a sterilization gas supply / recovery unit 50 for supplying sterilization gas to the transport module 2 and the work module 4.
  • the sterilization gas supply / recovery unit 50 may be detachable from the transport module 2 so that it can be connected (incorporated) to the cell culture system 1 only when sterilization is performed.
  • the transfer module 2 is provided with one continuous main gas line 10.
  • the main gas line 10 is connected to the sterilization gas supply / recovery unit 50 from an inlet end 10a serving as a gas supply connection port, and is an outlet serving as a gas discharge connection port connected to the sterilization gas supply / recovery unit 50. It extends continuously until it reaches the end 10b.
  • a normally open (open when not energized) on-off valve (dividing on-off valve) 10V is provided in the middle of the main gas line 10, and the main gas line 10 is upstream of the first region with the on-off valve 10V as a boundary. 11 and the second region 13 on the downstream side.
  • the first area 11 has a role as a main supply line
  • the second area 13 has a role as a main discharge line.
  • the transfer module 2 is provided with a plurality of branch supply lines 12 branched from the first region 11 of the main gas line 10.
  • the branch supply line 12 is for distributing the gas flowing through the first region 11 to each work module 4 connected to the transfer module 2.
  • the transfer module 2 has a plurality of branch discharge lines 14 branched from the second region 13 of the main gas line 10.
  • the branch discharge line 14 is for allowing the gas discharged from each work module 4 connected to the transfer module 2 to flow into the second region 13.
  • the transport module 2 has a main pressure measurement line 15 for measuring the pressure in each work module 4 and a plurality of branch pressure measurement lines 16 branched from the main pressure measurement line 15.
  • the pressure in each work module 4 is transmitted to the pressure gauge 59 provided in the sterilization gas supply / recovery unit 50 via the corresponding branch pressure measurement line 16 and the main pressure measurement line 15.
  • Each work module 4 is also provided with a pressure gauge 19 for measuring the pressure in each work module 4.
  • Each work module 4 is provided with a relief line 4L provided with a relief valve 4RV in order to release the pressure when the pressure in the internal space of the work module 4 becomes too high.
  • the transfer module 2 includes a main exhaust line 17 for sending the gas discharged from each work module 4 through the relief line 4L to the abatement device 70, and a plurality of branch exhaust lines 18 branched from the main exhaust line 17. Have. Exhaust gas rendered harmless by the abatement apparatus 70 can be discharged into an exhaust system set outside the cell culture system 1 such as a building such as a research facility.
  • the relief line 4L may be connected to the second region 13 of the main gas line 10, that is, the main discharge line.
  • the main exhaust line 17 may not be provided.
  • branch supply line 12, branch discharge line 14, and branch pressure measurement line 16 are provided with normally closed (non-energized) on-off valves 12V, 14V, and 16V, respectively.
  • the transfer module 2 Since the transfer module 2 is also a sterilization target module, the transfer module 2 includes various lines (branch supply line 12, branch discharge line 14, branch pressure measurement line 16, branch exhaust line 18 and the like) leading to the transfer space of the transfer module 2. Equivalent).
  • the transfer module 2 is provided with a relief line with a relief valve (equivalent to 4L and 4RV) communicating with the transfer space of the transfer module 2, and a pressure gauge 19 is also provided.
  • the plate loader module 3 is provided in the case where it has a chamber that is isolated from an external environment such as a load lock module and can be adjusted in atmosphere.
  • the above-mentioned various lines and pressure gauges are not provided.
  • the plate loader module 3 is a special module different from the work module 4, and the work module 4 can be attached to an attachment portion (mount, attachment port, etc.) of the plate loader module 3 in the transport module 2. Not.
  • the transfer module 2 (the transfer module part 2A and the transfer module part 2B) has a generally hollow rectangular parallelepiped or cubic shape as a whole. Each work module 4 also has a generally hollow rectangular parallelepiped or cubic shape. As shown in FIG. 2, the transport module 2 is provided with a plurality of connection ports 2 ⁇ / b> P (four are shown in the figure) for connecting the work module 4. The work module 4 is provided with a connection port 4P that can be airtightly connected to the connection port 2P of the transfer module 2.
  • connection port 2P of the transfer module 2 has the same structure and dimensions.
  • the connection ports 4P of each work module 4 have the same structure and dimensions regardless of the type of the work module 4. Therefore, a system having a function corresponding to a user's request can be constructed by combining the transport module 2 with a desired type of work module 4.
  • the interconnection between the two transfer module parts 2A and 2B can be performed, for example, by screwing the flanges together with the seals sandwiched between the flanges provided on the opposing end surfaces of the two transfer module parts.
  • the connection between the transport module 2 and the work module 4 for example, the flanges are screwed together with a seal interposed between the flange provided in the connection port 10P of the transport module 2 and the flange provided in the work module 4.
  • the male connector provided in one module and the female connector provided in the other module may be connected, and corresponding lines may be connected.
  • Guide rails 81 are provided on the floor surfaces of the transport module portions 2A and 2B. Guided by the guide rail 81, the base 83 of the medium plate transport robot 82 travels.
  • the medium plate transport robot 82 can be constituted by, for example, a horizontal articulated robot, and the pick (medium plate holding member) at the tip thereof can be directed in any direction within a horizontal plane and can move in any direction. Can be moved up and down.
  • the transfer module 2 is configured by connecting the two transfer module portions 2A and 2B, the two guide rails 81 are also connected so that the base 83 can travel from end to end of the transfer module 2. *
  • connection port 2P of the transfer module 2 is provided with a gate valve 85.
  • the gate valve 85 By opening the gate valve 85, the culture medium plate can be carried in and out of the work module 4 through the corresponding connection ports 2P and 4P.
  • the plate loader module 3 when the plate loader module 3 is an open special module, the plate loader module 3 can be connected to the transport module 2 by a connection structure different from the work module 4. In this case, it is not necessary to provide the gate valve 85 in the portion where the plate loader module 3 is attached.
  • the transfer module 2 may be configured by one module part. It is also possible to configure the transfer module 2 by combining three or more transfer module portions.
  • the configuration of each work module 4 will be briefly described with reference to FIG.
  • the work module 4 includes a chamber 41, work equipment 42 provided in the chamber, and a partition wall 44 that defines a sterilization management area 43 that is a part of the internal space of the chamber 41.
  • a part of the partition wall 44 may be movable.
  • the above-described relief line 4L in which the above-described relief valve 4RV is interposed is connected.
  • the relief line 4L is connected to the branch exhaust line 18 of the transport module 2.
  • the work equipment 42 provided in the work module 4 differs depending on the type of the work module 4.
  • the working device 42 includes a member that holds a medium plate, a device that adjusts the culture environment such as the temperature, humidity, and carbon dioxide gas concentration in the chamber, each device that performs medium adjustment, cell removal, and a microscope. Examples include observation equipment.
  • the work module 4 has a gas supply line 45.
  • the gas supply line 45 is connected to the branch supply line 12 of the transport module 2.
  • the gas supply line 45 opens in the sterilization management area 43 and discharges a gas such as sterilization gas into the sterilization management area 43.
  • the sterilization management area 43 is not completely isolated from the surrounding space in the chamber 41 (the surrounding chamber space 41 ′), but partly communicates with the surrounding chamber space 41 ′. . That is, the partition wall 44 does not completely surround the sterilization management area 43. Accordingly, when the sterilization gas is supplied into the sterilization management area 43, the surrounding chamber inner space 41 'is also filled with the sterilizing gas, and the surrounding chamber inner space 41' is also sterilized.
  • a gas discharge line 46 is opened in the surrounding chamber inner space 41 ′.
  • the gas discharge line 46 is connected to the branch discharge line 14 of the transport module 2. Therefore, the gas in the chamber 41 can be discharged to the outside of the chamber 41 through the gas discharge line 46.
  • An FFU (fan filter unit) 48 is provided on the ceiling of the partition wall 44.
  • the FFU 48 has a fan and a particle filter such as an ULPA filter (Ultra Low Low Penetration Air Air Filter), and filters and discharges the gas present in the surrounding chamber inner space 41 ′ into the sterilization management area 43. Thereby, the particle level of the gas in the chamber 41 can be kept below a certain level.
  • ULPA filter Ultra Low Low Penetration Air Air Filter
  • reference numeral 47 is a pressure detection line communicating with the branch pressure measurement line 16.
  • the sterilization management area 43 partitioned by the partition wall 44 may be used as the sterilization management area. Further, it may be difficult to provide the FFU in the chamber 41 in the relatively small work module 4 (however, the size of the connection port 4P is the same).
  • a circulation line in which a pump, a filter, a particle counter, and the like are connected to the chamber 41 (all are not shown), and the gas taken out from the chamber 41 is returned to the chamber 41 through the circulation line. The particles may be removed by a filter.
  • the sterilization gas supply / recovery unit 50 is one of sterilization gas hydrogen peroxide vapor, dry air (dry air) and inert gas, or a mixture of two or more.
  • the gas can be supplied to the inlet end 10 a of the main gas line 10. Further, the sterilization gas supply / recovery unit 50 can recover the exhaust gas discharged from the outlet end 10 b of the main gas line 10.
  • the sterilization gas supply / recovery unit 50 has a gas delivery line 51 connected to the inlet end 10 a of the main gas line 10.
  • the gas delivery line 51 is provided with a flow rate control mechanism 52 and an emergency shutoff valve 53.
  • Dry air or inert gas is supplied to the gas delivery line 51 from an external gas supply mechanism 54 connected to the sterilization gas supply / recovery unit 50.
  • a hydrogen peroxide vapor generator 55 is connected to the gas delivery line 51.
  • the flow rate of the gas (for example, dry air) supplied from the gas supply mechanism 54 is supplied by the flow rate control mechanism 52, and the hydrogen peroxide vapor supplied from the hydrogen peroxide vapor generator 55 is added to this flow rate-controlled gas flow.
  • a sterilized gas having a predetermined hydrogen peroxide concentration is sent out to the main gas line 10. That is, the sterilization gas supply / recovery unit 50 receives from the gas delivery line 51 a mixed gas containing air (preferably dry air) (which may be an inert gas) and hydrogen peroxide as sterilization gas, and hydrogen peroxide gas. Any one of air (which may be an inert gas) not included is selectively supplied.
  • air preferably dry air
  • hydrogen peroxide hydrogen peroxide
  • the sterilization gas supply / recovery unit 50 has a gas recovery line 56 connected to the outlet end 10 b of the main gas line 10.
  • the gas recovery line 56 is provided with a detoxifying mechanism 57 for detoxifying components (for example, hydrogen peroxide) harmful to the human body contained in the exhaust gas.
  • the gas recovery line 56 is provided with a hydrogen peroxide concentration meter 58 for measuring the hydrogen peroxide concentration in the gas flowing through the gas recovery line 56.
  • the gas recovery line 56 is connected to the gas delivery line 51 via the flow rate control mechanism 52.
  • the exhaust gas recovered from the main gas line 10 is detoxified by the detoxification mechanism 57 and returned to the gas delivery line 51, and the hydrogen peroxide vapor supplied from the hydrogen peroxide vapor generator 55 is added to this gas. Then, a sterilized gas having a predetermined hydrogen peroxide concentration is sent to the main gas line 10.
  • a blower (not shown) is provided in the gas delivery line 51 in order to give the gas a driving force for sending the gas returned to the gas delivery line 51 to the main gas line 10 again.
  • a dryer (not shown) is connected to the gas delivery line 51 from the connection point of the hydrogen peroxide vapor generator 55 in order to reduce the humidity of the gas. It is interposed at an upstream position.
  • the emergency shut-off valve 53 is closed to shut off the supply of sterilization gas when a dangerous situation such as an increase in the hydrogen peroxide concentration in the room where the cell culture system 1 is installed due to a leak.
  • the sterilization gas supply / recovery unit 50 further has a pressure gauge 59.
  • the pressure gauge 59 is connected to a connection port of the main pressure measurement line 15 provided in the transfer module 2, and the pressure in each work module 4 of the transfer module 2 through the main pressure measurement line 15 and the branch pressure measurement line 16. Is detected.
  • the pressure gauge 59 may be provided outside the sterilization gas supply / recovery unit 50.
  • a cassette (conveying container) containing a plurality of medium plates is placed on the plate loader module 3, and the conveying robot 82 takes out the medium plate (culture container) from the conveying container and conveys it to each work module 4 to each work module. 4, various operations such as preparation necessary for cell culture, observation of cultured cells, and removal of cultured cells are performed according to a predetermined schedule.
  • the medium plate holding the cultured cells is returned to the cassette of the plate loader module 3.
  • a description of the specific content of the work performed in each work module 4 is omitted.
  • the sterilization level inside the chamber 41 of each work module 4 (at least inside the sterilization management area 43 when the sterilization management area 43 is set) is Must meet predetermined criteria.
  • the sterilization level inside the transport module 2 that transports the culture plate to each work module 4 must also satisfy a predetermined standard. Further, if the medium plate is exposed to the ambient atmosphere in the plate loader module 3, the plate loader module 3 needs to be sterilized.
  • one module selected from the plurality of modules 2, 3 and 4 (hereinafter referred to as “sterilization target module”) is sterilized. That is, different modules are sterilized at different periods. Although it is possible to sterilize two modules in succession, it is preferred that only one module is sterilized at the same time.
  • the sterilization process is automatically performed according to the following procedure under the control of the system controller 101 that controls the operation of various devices constituting the cell culture system 1.
  • detection values of various measuring devices pressure gauges 19 and 59, a thermometer, a hygrometer, a carbon dioxide gas concentration sensor, etc.
  • the sterilization target module is one of the work modules 4 (hereinafter also referred to as “sterilization target module 4”).
  • CI chemical indicator
  • BI biological indicator
  • the open / close valve (supply open / close valve) 12V of the branch supply line 12 corresponding to the sterilization target module 4 is opened, and is supplied from the sterilization gas supply / recovery unit 50 via the first region 11 of the main gas line 10 and the branch supply line 12. Then, dry air not containing hydrogen peroxide vapor is supplied into the sterilization target module 4. At this time, the on-off valve of the branch discharge line 14 corresponding to the sterilization target module 4 is closed.
  • the sterilization target module 4 is filled with air until the detected value of the pressure gauge 19 provided in the sterilization target module 4 reaches a predetermined pressure (pressure lower than the set pressure of the relief valve 30R), and then the open / close valve of the branch supply line 12 Close 12V. After the elapse of a predetermined time, if the pressure drop calculated based on the detection value of the pressure gauge 19 is equal to or less than a predetermined threshold value, it is determined that the sealing property of the sterilization target module 4 is not a problem.
  • the pressure gauge 19 is also used for pressure management in the work module when performing various operations before and after culturing.
  • the gas that has returned to the gas recovery line 56 of the sterilization gas supply / recovery unit 50 flows again into the gas delivery line 51 and is dehumidified by a dryer (not shown) provided in the gas delivery line 51, and then the main gas line 10. To be supplied.
  • the dry air is supplied to the gas delivery line 51, the first region 11 of the main gas line 10, the branch supply line 12 corresponding to the sterilization target module 4, the sterilization target module 4, the branch discharge line 14 corresponding to the sterilization target module 4,
  • a chamber including the sterilization management area 43 of the sterilization target module 4 by circulating through a circulation path (hereinafter also referred to as “first circulation path”) including the second region 13 of the main gas line 10 and the gas recovery line 56. 41 is dehumidified.
  • sterilization of the sterilization management area 43 of the sterilization target module 4 is performed by flowing through the sterilization management area 43 when the sterilization gas circulates through the first circulation path.
  • the pressure in the sterilization target module 4 during the sterilization process is constantly monitored by the pressure gauge 59 of the sterilization gas supply / recovery unit 50, and the flow rate is set so that the pressure in the module 30 is maintained within a predetermined pressure range.
  • the control mechanism 52 adjusts the sterilization gas supply flow rate from the sterilization gas supply / recovery unit 50.
  • the pressure in the sterilization target module 4 is propagated to the pressure gauge 59 via the branch pressure measurement line 16 and the main pressure measurement line 15 by opening the on-off valve 18V of the branch pressure measurement line 16 corresponding to the sterilization target module 4. . Therefore, the pressure in the sterilization target module 4 can be monitored by the pressure gauge 59 located away from the sterilization target module 4.
  • the hydrogen peroxide concentration meter 58 constantly monitors the hydrogen peroxide concentration in the sterilization gas flowing through the gas recovery line 56. Based on the detection result of the hydrogen peroxide concentration meter 58, the flow rate of hydrogen peroxide vapor sent to the gas delivery line 51 from the hydrogen peroxide vapor generator 55 (that is, the hydrogen peroxide / air mixture ratio) is controlled.
  • the sterilization target module is in the sterilization process because there is a possibility that the sterilization gas from the module to be sterilized is slightly leaked from the seal part of the gate valve to the transport module In this case, it is desirable not to open the gate valve of another module. In this case, the culture container cannot be transported or collected to each module, but the processing in each module can be continued.
  • the gas passes through the gas delivery line 51, the main gas line 10 (the entire area of the first region 11 and the entire area of the second region 13), and the gas recovery line 56 (hereinafter also referred to as “second circulation path”). Air will circulate.
  • the following two parts that have become dead spaces that is, from the branch point of the branch supply line 12 connected to the sterilization target module 4 in the first region 11 of the main gas line 10 to the on-off valve 10V.
  • the sterilization gas staying in the part and the part from the branch point of the branch discharge line 14 connected to the module 4 to be sterilized in the second region 13 to the on-off valve 10V is forced from the part by air. Extruded and circulated through the second circulation path with air, during which the hydrogen peroxide in the sterilization gas is broken down into oxygen and water.
  • a gas line purge is performed. The process ends.
  • the aeration process and the main gas line purge process may be integrated. That is, when performing the aeration process, the on-off valve 10V of the main gas line 10 may be opened. By doing so, air flows through both the first circulation path and the second circulation path, and the internal spaces of the first circulation path and the second circulation path can be purged with air at the same time (the gas at this time (Refer to the thick line in FIG. 6 for the flow.)
  • the effect on sterilization gas can be confirmed by BI, that is, whether the bacteria contained in BI are killed by the gas, but on the other hand, it is necessary to check sterility regularly.
  • the agar medium may be transported, the falling bacteria may be collected and cultured, and the presence of bacteria may be confirmed. If the agar medium is brought into the target module and left for a certain period of time, if there are bacteria, it falls to the surface of the agar medium. The presence or absence of bacteria can be confirmed by growing and cultivating this for a certain period of time and removing it from the apparatus for inspection. Proliferation culture may be performed in an incubator outside the apparatus.
  • the open / close valve 10V is provided in the middle of one continuous main gas line 10, and the open / close valve 10V is closed during the sterilization process, whereby the main gas line 10 is connected to the gas supply line (first region). 11) and a gas exhaust line (second region 13).
  • the on / off valve 10V is opened with all of the on / off valves 12V and 14V of the branch supply line 12 and the branch discharge line 14 closed, and a purge gas (for example, dry air) is supplied from one end of the main gas line 10 to A purge gas can be allowed to flow from one end of 10 to the other end. For this reason, almost no sterilization gas remains in the main gas line 10. For this reason, it can prevent that the atmosphere in the laboratory where the cell culture apparatus was installed is contaminated by releasing the sterilized sterilization gas and harming a researcher, a worker, and the like.
  • a purge gas for example, dry air
  • the branch supply line 12 and the branch discharge line 14 each branching from the main gas line 10 and provided with the on-off valves 12V and 14V are connected to the sterilization target module (2, 3, 4). Since air supply and exhaust are performed, a plurality of modules to be sterilized can be sequentially and efficiently sterilized by one sterilization gas supply / recovery unit 50 by switching the on-off valves (12V, 14V).
  • the sterilization target module (2, 3, 3) is branched from the main pressure measurement line 15 through the branch pressure measurement line 16 provided with an on-off valve (pressure measurement on-off valve) 16V. 4)
  • the internal pressure can be measured at a remote location. For this reason, the gas supply state from the sterilization gas supply / recovery unit 50 can be easily adjusted based on the detection value of the pressure gauge 59 installed at a position away from the sterilization target module (2, 3, 4). .
  • the sterilization gas supply / recovery unit 50 sends the sterilization gas returned after being supplied to the sterilization target module to the delivery line 51 again, but is not limited thereto.
  • the sterilized gas that has returned may be discarded into an exhaust system set outside the cell culture system 1.

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  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Dans la présente invention, une conduite de gaz principale (10) mono-système installée dans un module de transport (2) est divisée en une première région (11) et une seconde région (13) par une vanne d'ouverture/fermeture de division (10V). Une pluralité de piquages d'alimentation (12) partent en parallèle de la première région et alimentent indépendamment en gaz des modules fonctionnels correspondants. Une pluralité de piquages d'évacuation (14) partent en parallèle de la seconde région et évacuent indépendamment le gaz des modules fonctionnels vers la conduite de gaz principale. Chacun des piquages d'alimentation est pourvu d'une vanne d'ouverture/fermeture d'alimentation (12V) et chacun des piquages d'évacuation est pourvu d'une vanne d'ouverture/fermeture d'évacuation (14V).
PCT/JP2016/085234 2015-11-27 2016-11-28 Système de culture cellulaire et procédé de stérilisation WO2017090773A1 (fr)

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JP2015232214A JP2019013151A (ja) 2015-11-27 2015-11-27 細胞培養システム
JP2015-232214 2015-11-27

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KR102596110B1 (ko) 2019-10-22 2023-10-30 바이오스페릭스 엘티디 화학적 살생물제를 사용하지 않는 무균 세포 처리 및 제조

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011030655A (ja) * 2009-07-30 2011-02-17 Sanyo Electric Co Ltd 滅菌庫
WO2015166554A1 (fr) * 2014-04-30 2015-11-05 株式会社エアレックス Système de décontamination

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011030655A (ja) * 2009-07-30 2011-02-17 Sanyo Electric Co Ltd 滅菌庫
WO2015166554A1 (fr) * 2014-04-30 2015-11-05 株式会社エアレックス Système de décontamination

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