WO2021049117A1 - Dispositif de culture cellulaire - Google Patents

Dispositif de culture cellulaire Download PDF

Info

Publication number
WO2021049117A1
WO2021049117A1 PCT/JP2020/023464 JP2020023464W WO2021049117A1 WO 2021049117 A1 WO2021049117 A1 WO 2021049117A1 JP 2020023464 W JP2020023464 W JP 2020023464W WO 2021049117 A1 WO2021049117 A1 WO 2021049117A1
Authority
WO
WIPO (PCT)
Prior art keywords
stirring
container
culture
cell suspension
containers
Prior art date
Application number
PCT/JP2020/023464
Other languages
English (en)
Japanese (ja)
Inventor
一弘 逵
山田　貴之
竹内　晴紀
Original Assignee
シンフォニアテクノロジー株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by シンフォニアテクノロジー株式会社 filed Critical シンフォニアテクノロジー株式会社
Publication of WO2021049117A1 publication Critical patent/WO2021049117A1/fr

Links

Images

Classifications

    • 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
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present invention relates to a cell culture device capable of moving a cell suspension between a plurality of culture vessels.
  • a work called "passage” is performed in which a cell suspension containing cells collected from a culture vessel is adjusted to a desired cell density and then transferred to a new culture vessel.
  • the conventional passage first, the cells adhering to the inner surface of the culture vessel are exfoliated with a stripping solution, the stripped cells are collected in a centrifuge tube, centrifuged, and the supernatant is removed, and then the liquid medium is used. Was added to prepare a cell suspension. Then, a part of the cell suspension was taken out, the number of cells was counted by a counting device, and a medium was appropriately added according to the measurement result to adjust the cell density of the cell suspension.
  • Patent Documents 1 and 2 disclose a robot arm that performs a series of operations related to such a passage.
  • Patent Documents 1 and 2 have a device configuration in which an isolator for forming a sterile space is provided and a robot arm is arranged in the sterile space.
  • an isolator is required in addition to the robot arm, there is a problem that the device becomes very expensive.
  • the present invention has been made in view of the above-mentioned problems, and an object of the present invention is to enable good stirring of a cell suspension with an inexpensive configuration in a cell culture device that does not use a centrifuge.
  • a plurality of culture containers in a sterile state are connected via a connecting tube while maintaining the sterile state, and the cell suspension is moved between the plurality of culture containers.
  • It is a cell culture device capable of connecting the plurality of stirring containers which are connected to the connecting tube and whose inside is in a sterile state, and the plurality of stirring containers while maintaining the sterile state of the plurality of stirring containers. It is characterized by including a stirring connection tube for moving the cell suspension and a driving unit for moving the cell suspension between the plurality of stirring containers via the stirring connection tube.
  • the cell suspension by moving the cell suspension between a plurality of stirring containers by the driving unit, the cell suspension can be greatly flowed, so that the cell suspension can be satisfactorily stirred. Further, since a so-called closed system configuration in which a plurality of culture containers and a plurality of stirring containers are connected in an aseptic state is adopted, it is not necessary to arrange the entire apparatus in an aseptic space, and an isolator is not required. Therefore, according to the present invention, the cell suspension can be satisfactorily agitated with an inexpensive structure.
  • the driving unit can move the cell suspension from the stirring container to another stirring container by pressure-feeding a predetermined gas to one of the plurality of stirring containers. It should be a possible gas supply unit.
  • a pump is not required to move the cell suspension, so that the cells are not crushed when the cell suspension passes through the pump.
  • a counting device connected to the connecting tube and counting the number of cells contained in the cell suspension is further provided, and the driving unit is used to select the stirring container from any of the plurality of stirring containers. It is advisable to move a part of the cell suspension to the counting device.
  • the cell suspension stirred by a plurality of stirring containers can be moved to the counting device without human intervention, so that the workload of the operator can be reduced.
  • the cell culture device 1 is a device for automatically culturing cells, and as shown in FIG. 1, includes a refrigerating storage unit 2, a culture unit 3, and a control unit 4.
  • the refrigerated storage unit 2 is a part for refrigerating and storing a medium tank and a stripping liquid tank (not shown), and the inside is maintained at a temperature lower than room temperature.
  • the medium tank contains a liquid medium necessary for cell culture.
  • the stripping solution tank contains a stripping solution for stripping cells adhering to the inner surface of the culture vessel.
  • the medium tank is connected to the medium container 15 (see FIG. 2) by a tube (not shown), and the medium tank is replenished with the medium from the medium tank 15 as needed.
  • the stripping liquid tank is connected to the stripping liquid container 16 (see FIG. 2) by a tube (not shown), and the stripping liquid tank 16 is replenished with the stripping liquid as needed.
  • the culture unit 3 is a site for performing cell culture, and the inside is maintained in a predetermined state suitable for cell culture (for example, temperature 37 ° C., humidity 95%, CO 2 concentration 5%).
  • Culture containers 11 and 12 are arranged inside the culture unit 3, and the cells in the culture containers 11 and 12 grow in a state of being attached to the inner surface of the container.
  • a door 3a is provided on the front surface of the culture unit 3, and cells can be taken out from the culture unit 3, parts in the culture unit 3 can be replaced, and the like.
  • the culture circuit 10 shown in FIG. 2 is formed in the cell culture device 1.
  • the culture circuit 10 includes a first culture container 11, a second culture container 12, a first stirring container 13, a second stirring container 14, a medium container 15, a stripping solution container 16, a counting device 17, and a counting device.
  • the waste liquid container 18 for the device, the waste liquid container 19, and the gas supply unit 20 are connected in a sterile state by tubes 31 to 39 to form a closed system.
  • an isolator can be eliminated.
  • the parts constituting the culture circuit 10 at least the first culture container 11, the second culture container 12, the first stirring container 13, the second stirring container 14, the medium container 15, and the stripping solution container 16 are contained in the culture unit 3. Is located in.
  • the first culture container 11 and the second culture container 12 are containers for performing cell culture, and the inside is maintained in an aseptic state.
  • a liquid medium is placed in the first culture container 11 and the second culture container 12, and the cells grow while adhering to the inner surface of the container in the medium.
  • a multi-layered container in which a plurality of culture layers are formed is adopted, but a container other than the multi-layered container may be used.
  • the first culture container 11 and the second culture container 12 are connected to the gas supply unit 20 by a tube 31.
  • the tube 31 is provided with a valve V1 for switching between supply and non-supply of gas from the gas supply unit 20 to the first culture container 11, and also supplies gas from the gas supply unit 20 to the second culture container 12.
  • a valve V2 for switching non-supply is provided.
  • the first culture vessel 11 and the second culture vessel 12 are connected by a tube 32 while maintaining an aseptic state.
  • the tube 32 is provided with a valve V3 for switching between supply and discharge of fluid to and from the first culture vessel 11 via the tube 32, and is provided to the second culture vessel 12 via the tube 32.
  • a valve V4 for switching between supply and discharge of fluid and non-supply and discharge of fluid is provided.
  • the first culture vessel 11 is provided with a degassing portion 52 having a valve V5 and a filter 51.
  • the second culture vessel 12 is provided with a degassing portion 54 having a valve V6 and a filter 53.
  • the first stirring container 13 and the second stirring container 14 are containers for stirring the cell suspension, as will be described in detail later.
  • the bag is adopted as the first stirring container 13 and the second stirring container 14, but a container other than the bag may be used.
  • the first stirring container 13 and the second stirring container 14 are connected to the gas supply unit 20 by a tube 31.
  • the tube 31 is provided with a valve V7 for switching between supply and non-supply of gas from the gas supply unit 20 to the first stirring container 13, and also supplies gas from the gas supply unit 20 to the second stirring container 14.
  • a valve V8 for switching non-supply is provided.
  • the first stirring container 13 and the second stirring container 14 are connected by a tube 33.
  • the tube 33 is connected to the tube 32 via the tube 34. That is, the first stirring container 13 and the second stirring container 14 are connected to the tube 32 via the tubes 33 and 34.
  • the tube 33 is provided with a valve V9, and the tube 34 is provided with a valve V10.
  • the first stirring container 13 is provided with a degassing portion 56 having a valve V11 and a filter 55.
  • the second stirring container 14 is provided with a degassing portion 58 having a valve V12 and a filter 57.
  • the medium container 15 is a container in which a liquid medium is stored.
  • the medium container 15 is connected to the gas supply unit 20 by a tube 31.
  • the tube 31 is provided with a valve V13 for switching between supply and non-supply of gas from the gas supply unit 20 to the medium container 15.
  • the medium container 15 is connected to the tube 32 via the tube 35.
  • the tube 35 is provided with a valve V14 for switching between supply and non-supply of the medium from the medium container 15.
  • the stripping liquid container 16 is a container in which the stripping liquid is stored.
  • the stripping liquid container 16 is connected to the gas supply unit 20 by a tube 31.
  • the tube 31 is provided with a valve V15 for switching between supply and non-supply of gas from the gas supply unit 20 to the stripping liquid container 16.
  • the stripper container 16 is connected to the tube 32 via the tube 36.
  • the tube 36 is provided with a valve V16 for switching between supply and non-supply of the release liquid from the release liquid container 16.
  • the counting device 17 is a device for counting the number of cells contained in the cell suspension.
  • the counting device 17 is connected to the tube 32 via the tube 37.
  • the tube 37 is provided with two valves V17 and V18.
  • a tube 38 is branched from a portion of the tube 37 between the valve V17 and the valve V18, and a waste liquid container 18 for a counting device is connected to the end of the tube 38.
  • the tube 38 is provided with a valve V19.
  • the waste liquid container 18 for the counting device is provided with a degassing portion 61.
  • the waste liquid container 19 is a container for discharging unnecessary liquid in the culture circuit 10.
  • the waste liquid container 19 is connected to the tube 37 via the tube 39. That is, the waste liquid container 19 is connected to the tube 32 via the tubes 39 and 37.
  • the tube 39 is provided with a valve V20.
  • the waste liquid container 19 is provided with a degassing portion 63 having a filter 62.
  • the gas supply unit 20 is a portion that supplies a predetermined gas (for example, air having a temperature of 37 ° C., a humidity of 95%, and a CO 2 concentration of 5%) that does not interfere with cell culture.
  • a predetermined gas for example, air having a temperature of 37 ° C., a humidity of 95%, and a CO 2 concentration of 5%.
  • the culture circuit 10 is configured so that the liquid in each container 11 to 16 can be moved by pumping gas from the gas supply unit 20.
  • the gas supply unit 20 has a CO 2 cylinder 71 and an air cylinder 72. From CO 2 cylinder 71, the pressure is adjusted by the regulator 73, and, CO 2 whose flow rate is adjusted by the mass flow controller 74 is supplied. From the air cylinder 72, air whose pressure is adjusted by the regulator 75 and whose flow rate is adjusted by the mass flow controller 76 is supplied. CO air supplied from the CO 2 and Eabonbe 72 supplied from the 2 cylinder 71 are mixed, the CO 2 concentration in the mixed gas is detected by the CO 2 sensor 77. The control unit 4 controls the mass flow controllers 74 and 76 so that the CO 2 concentration is maintained at 5% according to the output of the CO 2 sensor 77. The gas thus obtained is supplied to each of the containers 11 to 16 after bacteria and the like are removed by the filter 78.
  • the control unit 4 controls the opening and closing of the valves V1 to V20 and the supply of gas from the gas supply unit 20, so that the passage is automatically performed.
  • Various conditions related to culturing and subculture are input to the control unit 4 in advance by the operator.
  • the valves V1 to V20 are closed and the culture circuit 10 is maintained in an aseptic state.
  • the cells cultured in the first culture vessel 11 are collected, the cell suspension is adjusted to a desired cell density, and then the cells are transferred to the second culture vessel 12.
  • the control unit 4 opens the valves V1, V3, and V20, and pumps gas from the gas supply unit 20. Then, the medium M in the first culture container 11 is discharged to the waste liquid container 19 through the tubes 32, 37, 39. At this time, since the cells C are attached to the inner surface of the first culture vessel 11, they are not discharged together with the medium M.
  • the control unit 4 returns the valves V1, V3, and V20 to the closed state, and stops the gas supply from the gas supply unit 20.
  • the stripping solution L is supplied to the first culture vessel 11 (step S13, FIG. 6B). Specifically, the control unit 4 opens the valves V3, V5, V15, and V16, and pumps gas from the gas supply unit 20. Then, the stripping liquid L in the stripping liquid container 16 is supplied to the first culture vessel 11 through the tubes 36 and 32. When a predetermined amount of the stripping liquid L is supplied to the first culture vessel 11, the control unit 4 returns the valves V3, V5, V15, and V16 to the closed state, and stops the gas supply from the gas supply unit 20.
  • This first waiting time is the time until the cells C adhering to the inner surface of the first culture vessel 11 are in a state immediately before being exfoliated by the chemical action of the exfoliating liquid L.
  • the first waiting time is appropriately determined depending on the type of cell C and the type of exfoliating solution L, and is, for example, about 2 to 3 minutes.
  • the stripping solution L is discharged from the first culture vessel 11 (step S15, FIG. 6 (c)). Specifically, the control unit 4 opens the valves V1, V3, and V20, and pumps gas from the gas supply unit 20. Then, the stripping liquid L in the first culture container 11 is discharged to the waste liquid container 19 through the tubes 32, 37, 39. At this time, since the cells C are still attached to the inner surface of the first culture vessel 11, they are not discharged together with the stripping solution L. When the stripping liquid L in the first culture vessel 11 is discharged, the control unit 4 returns the valves V1, V3, and V20 to the closed state, and stops the gas supply from the gas supply unit 20. By discharging the stripping solution L before the cells C are separated in this way, it is not necessary to centrifuge the stripping solution L and the cells C later, so that a centrifuge can be eliminated.
  • the process waits for a predetermined second standby time in that state (step S16). Even if the stripping liquid L is discharged, the stripping liquid L actually remains slightly adhered to the inner surface of the first culture vessel 11 and the cells C due to surface tension or the like, as shown in FIG. 6D. ..
  • the second waiting time is the time until the cells C, which are in the state immediately before exfoliation, are completely exfoliated by the chemical action of the residual liquid of the exfoliating liquid L.
  • the second waiting time is appropriately determined depending on the type of cell C and the type of exfoliating solution L, and is, for example, about 2 to 3 minutes.
  • the medium M is supplied to the first culture vessel 11 (step S17). Specifically, the control unit 4 opens the valves V3, V5, V13, and V14, and pumps gas from the gas supply unit 20. Then, the medium M in the medium container 15 is supplied to the first culture container 11 through the tubes 35 and 32. As a result, as shown in FIG. 6 (e), the cells C exfoliated from the inner surface of the first culture vessel 11 are suspended in the medium M to prepare a cell suspension S. Since the residual liquid of the stripping liquid L is a small amount, it is sufficiently diluted by supplying the medium M, and there is no problem in a later step. When a predetermined amount of the medium M is supplied to the first culture vessel 11, the control unit 4 returns the valves V3, V5, V13, and V14 to the closed state, and stops the gas supply from the gas supply unit 20.
  • the cell suspension S in the first culture vessel 11 is moved to the first stirring vessel 13 (step S18). Specifically, the control unit 4 opens the valves V1, V3, V10, and V11, and pumps gas from the gas supply unit 20. Then, the cell suspension S in the first culture vessel 11 moves to the first stirring vessel 13 through the tubes 32 and 34. When the cell suspension S is collected in the first stirring container 13, the control unit 4 returns the valves V1, V3, V10, and V11 to the closed state, and stops the gas supply from the gas supply unit 20. Although the cell suspension S is collected in the first stirring container 13 here, the cell suspension S may be collected in the second stirring container 14.
  • the cell suspension is stirred (step S19). Specifically, the cell suspension S is agitated by moving the cell suspension S between the first stirring container 13 and the second stirring container 14. In the present embodiment, the cell suspension S is moved between the first stirring container 13 and the second stirring container 14 three times, but the number of times can be changed as appropriate and sufficient stirring can be performed. If so, it may be done once.
  • FIG. 7 is a schematic view for explaining the stirring operation of the cell suspension S, and shows only the configuration related to stirring. In FIG. 7, the valve painted in black is in the closed state, and the valve not painted in black is in the open state.
  • the control unit 4 opens the valves V7, V9, and V12, and pumps gas from the gas supply unit 20. Then, the cell suspension S in the first stirring container 13 moves to the second stirring container 14 through the tube 33. Subsequently, as shown in FIG. 7B, the control unit 4 returns the valves V7 and V12 to the closed state and opens the valves V8 and V11. Then, the cell suspension S in the second stirring container 14 moves to the first stirring container 13 through the tube 33. Further, as shown in FIG. 7A, the control unit 4 returns the valves V8 and V11 to the closed state and opens the valves V7 and V12. Then, again, the cell suspension S in the first stirring container 13 moves to the second stirring container 14 through the tube 33.
  • the cell suspension S flows greatly and is appropriately stirred.
  • the amount of the cell suspension S to be moved at the time of stirring may be the whole amount or a part, but finally, the whole amount of the cell suspension S is moved to either the first stirring container 13 or the second stirring container 14. Shall be made to.
  • the control unit 4 when the entire amount of the cell suspension S has moved to the second stirring vessel 14, the control unit 4 returns the valves V7, V9, and V12 to the closed state, and stops the gas supply from the gas supply unit 20. To do.
  • step S20 a part of the cell suspension S in the second stirring container 14 is moved to the counting device 17 (step S20). Specifically, the control unit 4 opens the valves V8, V9, V10, V17, and V18, and pumps gas from the gas supply unit 20. Then, the cell suspension S in the second stirring container 14 moves to the counting device 17 through the tubes 33, 34, 32, 37. When the predetermined amount of the cell suspension S moves to the counting device 17, the control unit 4 returns the valves V8, V9, V10, V17, and V18 to the closed state, and stops the gas supply from the gas supply unit 20.
  • the counting device 17 counts the number of cells contained in the trace amount of cell suspension S (step S21).
  • the measurement result by the counting device 17 is transmitted to the control unit 4.
  • the control unit 4 calculates an additional amount of medium required to bring the cell suspension S in the second stirring vessel 14 to a desired cell density. If the number of cells is too large, a part of the cell suspension S in the second stirring container 14 may be discharged before adding the medium.
  • the cell suspension S used in the counting device 17 is discharged into the waste liquid container 18 for the counting device after the measurement is completed.
  • the control unit 4 opens the valves V18 and V19 and operates a syringe pump (not shown) included in the counting device 17. As a result, the cell suspension S remaining in the counting device 17 is discharged to the waste liquid container 18 for the counting device through the tubes 37 and 38.
  • the control unit 4 adjusts the cell density of the cell suspension S by adding the amount of the medium obtained in the above calculation to the second stirring vessel 14 (step S22). Specifically, the control unit 4 opens the valves V9, V10, V12, V13, and V14, and pumps gas from the gas supply unit 20. Then, the medium M in the medium container 15 is supplied to the second stirring container 14 through the tubes 35, 32, 34, 33. When the required amount of medium is added to the second stirring container 14, the control unit 4 returns the valves V9, V10, V12, V13, and V14 to the closed state, and stops the gas supply from the gas supply unit 20.
  • step S23 After adjusting the cell density of the cell suspension S, the cell suspension S is stirred again (step S23). This is because there is a possibility that the cells C in the cell suspension S have settled in the second stirring vessel 14 due to the passage of time while the counting device 17 is counting the number of cells. However, if the cells C in the cell suspension S are uniformly dispersed in the second stirring vessel 14, step S23 can be omitted. Since the stirring operation is the same as in step S19, the explanation is omitted.
  • Step S24 the control unit 4 opens the valves V4, V6, V8, V9, and V10, and pumps gas from the gas supply unit 20. Then, the cell suspension S in the second stirring container 14 moves to the second culture container 12 through the tubes 33, 34, 32.
  • the control unit 4 returns the valves V4, V6, V8, V9, and V10 to the closed state, and stops the gas supply from the gas supply unit 20. In this way, the passage from the first culture vessel 11 to the second culture vessel 12 is completed.
  • the connected tubes may be replaced with new ones, or the inside of the tubes may be cleaned and reused.
  • a welding machine such as BioWelder (manufactured by Sartorius Stedim Japan) or OPTA sterile connector (manufactured by Sartorius Stedim Japan) is used, each container or device is replaced while maintaining the sterile condition inside the tube. It is also possible.
  • a plurality of culture containers 11 and 12 whose inside is sterile are connected via a tube 32 (corresponding to the connecting tube of the present invention) while maintaining the sterile state, and the plurality of culture containers 11 and 12 are connected.
  • the cell culture apparatus 1 capable of moving the cell suspension between, a plurality of stirring containers 13 and 14 connected to a tube 32 and having a sterile inside and a plurality of stirring containers 13 and 14 being sterile.
  • the cell suspension is moved between the tube 33 (corresponding to the stirring connection tube of the present invention) connecting the plurality of stirring containers 13 and 14 and the plurality of stirring containers 13 and 14 via the tube 33 while maintaining the state.
  • a gas supply unit 20 (corresponding to the drive unit of the present invention) is provided.
  • the cell suspension can be largely flowed by moving the cell suspension between the plurality of stirring vessels 13 and 14 by the gas supply unit 20, so that the cells can be largely flowed.
  • the suspension can be well stirred.
  • a so-called closed system configuration is adopted in which a plurality of culture vessels 11 and 12 and a plurality of stirring vessels 13 and 14 are connected in an aseptic state, it is not necessary to arrange the entire apparatus in an aseptic space, and the isolator can be used. It becomes unnecessary. Therefore, the cell suspension can be well stirred with an inexpensive configuration.
  • the driving unit moves a cell suspension from the stirring container to another stirring container by pressure-feeding a predetermined gas to any of the plurality of stirring containers 13 and 14. It is composed of a gas supply unit 20 capable of With such a configuration, a pump is not required to move the cell suspension, so that the cells are not crushed when the cell suspension passes through the pump.
  • the counting device 17 which is connected to the tube 32 and counts the number of cells contained in the cell suspension is further provided, and the gas supply unit 20 stirs any one of the plurality of stirring containers 13 and 14. It is configured to move a portion of the cell suspension from the container to the counting device 17. With such a configuration, the cell suspension stirred by the plurality of stirring containers 13 and 14 can be moved to the counting device 17 without human intervention, so that the work load on the operator can be reduced. it can.
  • the number of culture containers and the number of stirring containers are set to 2 each, but 3 or more may be provided.
  • the gas supply unit 20 is provided as a means (driving unit) for moving the cell suspension, the medium, and the exfoliating liquid, and the liquid is moved by pumping the gas.
  • the means for moving the cell suspension, the medium and the stripping solution is not limited to the gas supply unit 20, and a pump may be used, for example.
  • the number of cells is automatically counted by the counting device 17.
  • the agitated cell suspension may be taken out by the operator from the culture circuit 10 and the operator may count the number of cells with a counting device.
  • Cell culture device 11 First culture container (culture container) 12: Second culture container (culture container) 13: First stirring container (stirring container) 14: Second stirring container (stirring container) 17: Counting device 20: Gas supply unit (drive unit) 32: Tube (connecting tube) 33: Tube (stirring connection tube)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Virology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention a pour but de permettre une bonne agitation d'une suspension cellulaire à l'aide d'une structure peu coûteuse dans un dispositif de culture cellulaire sans centrifugation. L'invention concerne en outre un dispositif de culture cellulaire dans lequel une pluralité de récipients de culture (11) et (12) étant stériles à l'intérieur sont reliés par l'intermédiaire d'un tube de raccordement (32) tout en maintenant la stérilité de telle sorte qu'une suspension cellulaire peut être transférée entre la pluralité de récipients de culture (11) et (12), une pluralité de récipients d'agitation (13) et (14) qui sont stériles à l'intérieur et reliés au tube de raccordement (32), un tube de raccordement à agitation (33) reliant la pluralité de récipients d'agitation (13) et (14) tout en maintenant la stérilité de la pluralité de récipients d'agitation (13) et (14), et une unité d'entraînement (20) pour transférer la suspension de cellules entre la pluralité de récipients d'agitation (13) et (14) et par l'intermédiaire du tube de raccordement à agitation (33).
PCT/JP2020/023464 2019-09-12 2020-06-15 Dispositif de culture cellulaire WO2021049117A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2019166501A JP2021040575A (ja) 2019-09-12 2019-09-12 細胞培養装置
JP2019-166501 2019-09-12

Publications (1)

Publication Number Publication Date
WO2021049117A1 true WO2021049117A1 (fr) 2021-03-18

Family

ID=74862115

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2020/023464 WO2021049117A1 (fr) 2019-09-12 2020-06-15 Dispositif de culture cellulaire

Country Status (3)

Country Link
JP (1) JP2021040575A (fr)
TW (1) TW202115238A (fr)
WO (1) WO2021049117A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI795909B (zh) * 2021-09-16 2023-03-11 國立臺灣大學 自動化細胞培養裝置及自動化細胞培養系統

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005198626A (ja) * 2004-01-19 2005-07-28 Sanyo Electric Co Ltd 自動継代培養装置及びこれを用いた継代培養方法
JP2015188391A (ja) * 2014-03-28 2015-11-02 東洋製罐グループホールディングス株式会社 細胞培養方法、及び細胞培養システム
WO2016063364A1 (fr) * 2014-10-22 2016-04-28 株式会社日立ハイテクノロジーズ Mécanisme de cytométrie, dispositif de culture cellulaire le comprenant et procédé de cytométrie
JP2017006058A (ja) * 2015-06-23 2017-01-12 国立大学法人京都大学 培養装置及び培養方法
WO2019138796A1 (fr) * 2018-01-15 2019-07-18 富士フイルム株式会社 Dispositif de culture cellulaire et procédé de culture cellulaire
WO2020009017A1 (fr) * 2018-07-05 2020-01-09 富士フイルム株式会社 Dispositif de culture cellulaire et méthode d'agitation

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005198626A (ja) * 2004-01-19 2005-07-28 Sanyo Electric Co Ltd 自動継代培養装置及びこれを用いた継代培養方法
JP2015188391A (ja) * 2014-03-28 2015-11-02 東洋製罐グループホールディングス株式会社 細胞培養方法、及び細胞培養システム
WO2016063364A1 (fr) * 2014-10-22 2016-04-28 株式会社日立ハイテクノロジーズ Mécanisme de cytométrie, dispositif de culture cellulaire le comprenant et procédé de cytométrie
JP2017006058A (ja) * 2015-06-23 2017-01-12 国立大学法人京都大学 培養装置及び培養方法
WO2019138796A1 (fr) * 2018-01-15 2019-07-18 富士フイルム株式会社 Dispositif de culture cellulaire et procédé de culture cellulaire
WO2020009017A1 (fr) * 2018-07-05 2020-01-09 富士フイルム株式会社 Dispositif de culture cellulaire et méthode d'agitation

Also Published As

Publication number Publication date
TW202115238A (zh) 2021-04-16
JP2021040575A (ja) 2021-03-18

Similar Documents

Publication Publication Date Title
RU2644231C2 (ru) Автомат и автоматизированный способ культивирования клеток
US11261416B2 (en) Fixed-bed bioreactor with constant-flow pump / tubing system
JP6322711B2 (ja) 細胞数濃度調整装置およびそれを用いた自動継代培養システム
JP2020072684A (ja) 細胞懸濁液処理装置およびそれを用いた自動継代培養システム
EP3604547A1 (fr) Bioréacteur à lit fixe comportant un système de pompe/tube à débit constant
WO2021049117A1 (fr) Dispositif de culture cellulaire
JP4225924B2 (ja) 自動継代培養装置及びこれを用いた継代培養方法
US20200339930A1 (en) Cell culture device and cell culture method
JP2022506976A (ja) 細胞培養システムおよび方法
EP3988639A1 (fr) Procédé de récupération de cellules et dispositif de culture cellulaire
JP6858653B2 (ja) 送液装置およびそれを用いた細胞培養装置
JP2019162035A (ja) 細胞処理装置
JP7481608B2 (ja) 細胞培養装置
JP2019535010A (ja) 分離樹脂の移送のための方法およびシステム
JP2019000059A5 (fr)
WO2020189655A1 (fr) Système de culture, dispositif de culture, et dispositif de manipulation de récipient de culture multicouche
US20210355425A1 (en) Material transfer devices and related systems and methods
JP2020156465A (ja) 培養システム、培養装置、および多層培養容器操作装置
WO2012065618A1 (fr) Dispositif de passage des cellules

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20863442

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20863442

Country of ref document: EP

Kind code of ref document: A1