WO2015147077A1 - Procédé de fabrication d'un récipient de remplissage de cellules, récipient pour remplissage de cellules et dispositif d'étanchéité - Google Patents

Procédé de fabrication d'un récipient de remplissage de cellules, récipient pour remplissage de cellules et dispositif d'étanchéité Download PDF

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Publication number
WO2015147077A1
WO2015147077A1 PCT/JP2015/059189 JP2015059189W WO2015147077A1 WO 2015147077 A1 WO2015147077 A1 WO 2015147077A1 JP 2015059189 W JP2015059189 W JP 2015059189W WO 2015147077 A1 WO2015147077 A1 WO 2015147077A1
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WIPO (PCT)
Prior art keywords
storage chamber
storage
cell
filled
container
Prior art date
Application number
PCT/JP2015/059189
Other languages
English (en)
Japanese (ja)
Inventor
吉川　義洋
桂 松尾
智彦 内村
亮 富井
本望　修
Original Assignee
北海道公立大学法人札幌医科大学
ニプロ株式会社
吉川　義洋
桂 松尾
智彦 内村
亮 富井
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.)
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Application filed by 北海道公立大学法人札幌医科大学, ニプロ株式会社, 吉川　義洋, 桂 松尾, 智彦 内村, 亮 富井 filed Critical 北海道公立大学法人札幌医科大学
Priority to JP2016510438A priority Critical patent/JP6869455B2/ja
Publication of WO2015147077A1 publication Critical patent/WO2015147077A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0236—Mechanical aspects
    • A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0236—Mechanical aspects
    • A01N1/0263—Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
    • A01N1/0268—Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/02—Blood transfusion apparatus
    • A61M1/0209—Multiple bag systems for separating or storing blood components
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2207/00—Methods of manufacture, assembly or production
    • 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
    • C12M45/00—Means for pre-treatment of biological substances
    • C12M45/22—Means for packing or storing viable microorganisms

Definitions

  • the present invention relates to a production method for obtaining a cell-filled container by filling a cell-containing liquid into the container, a cell-filling container, and an apparatus for sealing the cell-filled container.
  • Patent Document 1 discloses a cell cryopreservation container in which a plurality of storage chambers are provided at intervals, and adjacent storage chambers communicate with each other through a communication portion. According to this cell cryopreservation container, since the cell suspension of stem cells is cryopreserved in a plurality of storage chambers, it can be thawed after being separated for each storage chamber.
  • the same cell suspension used for regenerative medicine is required to be stored as a test sample for quality control and the like.
  • the cell suspension in contact with the contaminated storage chamber needs to be stored as a test sample.
  • the plurality of storage chambers as described above are communicated by a communication path when the cell suspension is injected, and after the cell suspension is injected, the communication path is closed and each storage chamber is sealed.
  • a communication path is closed by heat welding. If the cell suspension is heated during this thermal welding, the cells may be damaged.
  • the present invention has been made in view of the circumstances described above, and an object thereof is to provide a method for producing a cell-filled container suitable for storing a liquid test sample containing cells, and a cell-filling container. .
  • Another object of the present invention is to provide a sealing means in which damage to cells is suppressed in a cell filling container for storing a liquid containing cells.
  • the present invention is connected to a first storage section that defines a first storage chamber having at least one port, via the first storage section and the first connection section, and has at least one port.
  • a container comprising: a second storage section that defines a second storage chamber; and at least two communication paths that are provided in the first connection section and communicate with the first storage chamber and the second storage chamber.
  • the present invention relates to a method for manufacturing a cell-filled container that obtains a cell-filled container by filling a liquid containing cells.
  • the method for producing the cell-filled container includes filling the liquid into the first storage chamber in a state where the liquid does not flow into the second storage chamber through the communication path until the first storage chamber is filled with the liquid. And a second step of allowing the liquid filled in the first storage chamber to flow into the second storage chamber through the communication path.
  • the liquid suitable for the test sample is stored in the second storage chamber.
  • the second storage chamber is positioned above the first storage chamber in the direction of gravity, and the liquid is filled in the first storage chamber.
  • the second storage chamber is The two storage chambers may be positioned below the first storage chamber in the direction of gravity, and the liquid filled in the first storage chamber may flow into the second storage chamber through the communication path.
  • At least the first storage chamber may be decompressed and then the liquid may be filled into the first storage chamber.
  • the liquid can be filled without allowing the first storage chamber to communicate with the atmosphere.
  • the communication path of the first connecting portion may be sealed after the second storage chamber is filled with the liquid.
  • the first storage chamber and the second storage chamber can be separated.
  • the volume of the second storage chamber may be smaller than the volume of the first storage chamber.
  • the communication path of the first connecting portion may be pressed and closed, and the pressed portion may be thermally welded and sealed.
  • a sealing member in which a heat generating member is disposed between the first heat insulating material and the second heat insulating material, on the first housing portion side and the second housing portion side of the communication path of the first connecting portion.
  • the first heat insulating material and the second heat insulating material may be arranged and pressed, and the first heat insulating material and the second heat insulating material may be thermally welded by the heat generating member.
  • the heat generating member is heated and thermally welded while the communication path is pressed and closed by the first heat insulating material and the second heat insulating material, there is no liquid containing cells in the heat welded portion, Heat at the time of welding is difficult to transfer to the liquid containing cells.
  • the container is connected to the opposite side of the first storage part via the second storage part and the second connection part, and a third storage that defines a third storage chamber having at least one port. And at least two communication passages that are provided in the second connection portion and communicate with the second storage chamber and the third storage chamber. In the second step, The liquid filled in the first storage chamber may flow into the third storage chamber.
  • the sum of the volume of the second storage chamber and the volume of the third storage chamber may be smaller than the volume of the first storage chamber.
  • the second storage chamber and the third storage chamber are easily filled with the liquid filled in the first storage chamber.
  • the cell filling container according to the present invention is connected via a first storage part that defines a first storage chamber having at least one port, and the first storage part and the first connection part, A second accommodating portion that defines a second accommodating chamber having at least one port, and is connected to the opposite side of the first accommodating portion via the second accommodating portion and the second connecting portion; and at least one port A third storage section that divides the third storage chamber, and at least two communication passages that are provided in the first connection section and communicate with the first storage chamber and the second storage chamber, and 2 at least two communication passages that are provided in the two connecting portions and communicate with the second storage chamber and the third storage chamber.
  • the sum of the volume of the second storage chamber and the volume of the third storage chamber is smaller than the volume of the first storage chamber.
  • the liquid filled in the first storage chamber can surely flow into the second storage chamber and the third storage chamber to be filled.
  • Each communication path provided in the first connection part and each communication path provided in the second connection part are along a first direction in which the first accommodation part and the second accommodation part are arranged. Each extending in parallel and parallel to a second direction orthogonal to the first direction, and the first accommodating portion, the second accommodating portion, and the third accommodating portion are arranged in the first direction and The length of the outer shape in the third direction orthogonal to the second direction is shorter than the length of the outer shape in the second direction, and each communication path provided in the first connecting portion, and the second connecting portion
  • the communication paths provided in the first and second storage portions, the second storage portion, and the third storage portion are arranged to be biased toward one side in the second direction with respect to the center position of the outer shape in the second direction. May be.
  • Each communication passage provided in the first connection portion and each opening provided in the second connection portion to the second storage chamber or the third storage chamber are connected to the cell.
  • the filling container In a state where the first container is placed above the gravitational direction and the third container is placed below the gravitational direction, the filling container is located at the uppermost position in the gravitational direction in the second container or the third container. You may do.
  • the present invention includes a first storage section that partitions the first storage chamber, and a second storage section that is connected to the first storage section via the first connection section and that partitions the second storage chamber.
  • a container that is provided in the first connecting portion and communicates with the first storage chamber and the second storage chamber is filled with a liquid containing cells, and the communication passage is sealed.
  • the present invention relates to a method for producing a cell-filled container.
  • the manufacturing method of a cell filling container includes the 1st process of pressing and closing the said communicating path, and the 2nd process of heat-welding the location where the said communicating path is pressed.
  • the heat-welded portion may be cut to separate the first housing portion and the second housing portion.
  • the first storage part and the second storage part can be stored in different places.
  • the communication path may be pressed by a heat insulating material.
  • the present invention is a liquid containing cells, which is connected to the first storage part that partitions the first storage chamber filled with the liquid containing cells, and the first storage part and the first connection part.
  • the second storage part that divides the second storage chamber filled with the liquid and the first connection part are provided so that the liquid containing the cells can communicate between the first storage chamber and the second storage chamber.
  • a sealing device for sealing a cell-filled container includes a first heat insulating material and a second heat insulating material that press and close the first housing portion side and the second housing portion side of the communication path, and the first heat insulating material and the second heat insulating material.
  • a heat generating member that is disposed between the materials and that heat-welds by pressing the communication passage.
  • the heat generating member is heated and thermally welded while the communication path is pressed and closed by the first heat insulating material and the second heat insulating material, there is no liquid containing cells in the heat welded portion, Heat at the time of welding is difficult to transfer to the liquid containing cells.
  • a liquid test sample containing cells filled in the first storage chamber can be stored in the second storage chamber.
  • FIG. 1 is a perspective view showing a container 10 according to an embodiment of the present invention.
  • FIG. 2 is a cross-sectional view showing the internal structure of the container 10.
  • FIG. 3 is a cross-sectional view showing the state of the container 10 in the first step.
  • FIG. 4 is a cross-sectional view showing a state where the cell suspension 40 is filled in the first storage chamber 14 in the first step.
  • FIG. 5 is a cross-sectional view showing the state of the container 10 in the second step.
  • FIG. 6 is a cross-sectional view showing a state in which the cell suspension 40 has flowed from the first storage chamber 14 to the second storage chamber 22 and the third storage chamber 27 in the second step.
  • FIG. 1 is a perspective view showing a container 10 according to an embodiment of the present invention.
  • FIG. 2 is a cross-sectional view showing the internal structure of the container 10.
  • FIG. 3 is a cross-sectional view showing the state of the container 10 in the first step.
  • FIG. 4 is
  • FIG. 7 is a cross-sectional view showing a state in which the communication passages 23, 24, 29, and 30 are sealed in the second step.
  • FIG. 8 is a cross-sectional view illustrating a state in which the first housing portion 11, the second housing portion 12, and the third housing portion 13 are separated in the second step.
  • FIG. 9 is a schematic diagram showing the main configuration of the sealing device 60.
  • FIG. 10 is a diagram illustrating a configuration of the first heater unit 63.
  • FIG. 11 is a view showing a state in which a part 43 of the first connecting portion 21 of the container 10 is pressed by the heat insulating materials 71 and 72.
  • FIG. 12 is a view showing a state in which the sealing portion 42 is formed in the first connecting portion 21 of the container 10.
  • the container 10 As shown in FIGS. 1 and 2, the container 10 (corresponding to a cell filling container) is for cryopreserving the cell suspension 40 (see FIG. 4).
  • the container 10 is configured, for example, by laminating two resin sheets molded into a shape described later.
  • the container 10 has a first housing part 11, a second housing part 12, and a third housing part 13.
  • the 1st accommodating part 11 has the 1st accommodating chamber 14 which is internal space, and the length of the left-right direction 52 (equivalent to a 1st direction) and the up-down direction 53 (equivalent to a 2nd direction).
  • the length of the front-rear direction 51 (corresponding to the third direction) is the shortest shape.
  • the first storage chamber 14 is a substantially rectangular parallelepiped space, the upper portion in the up-down direction 53 has the maximum length in the left-right direction 52, and the lower portion in the up-down direction 53 has the length in the left-right direction 52. It is curved into a funnel shape that gradually shortens downward.
  • the volume of the first storage chamber 14 is appropriately set in consideration of the amount of the cell suspension 40 to be stored, and is about 20 mL, for example.
  • the first storage unit 11 is provided with a cell injection port 15, a spare port 16, and a cell recovery port 17 that lead to the first storage chamber 14.
  • the cell injection port 15 is provided at the upper end in the up-down direction 53 of the first storage chamber 14 and in the vicinity of the left end in the left-right direction 52 (the left end of the first storage unit 11 in FIG. 1), and opens to the outside.
  • the first storage chamber 14 communicates with the outside.
  • a tube 18 is connected to the cell injection port 15 via a cylindrical member 33.
  • the cylindrical member 33 is a cylindrical member made of rubber or elastomer, and is inserted into the cell injection port 15 in a liquid-tight manner.
  • the cylindrical member 33 does not easily block the cell injection port 15 even if it receives an external force or the like or the first storage chamber 14 is decompressed.
  • the tube 18 is a resin tube. One end of the tube 18 is liquid-tightly connected to the cylindrical member 33. The cell suspension 40 is circulated through the tube 18, and the cell suspension 40 is injected into the first storage chamber 14 through the cell injection port 15.
  • the spare port 16 is provided at the upper end in the vertical direction 53 of the first storage chamber 14 and in the vicinity of the right end in the left-right direction 52 (the right end of the first storage portion 11 in FIG. 1).
  • the upper end of the spare port 16 is sealed and does not open to the outside.
  • the upper end side of the spare port 16 is cut and opened.
  • a cylindrical stopper 34 is inserted into the spare port 16 in a liquid-tight manner.
  • the plug 34 has a cylindrical cylindrical body and a film that divides the internal space of the cylindrical body into one end side (upper side in FIG. 2) and the other end side (lower side in FIG. 2) by rubber, elastomer, or the like. It is formed.
  • the spare port 16 Even if the spare port 16 receives an external force or the like or the first storage chamber 14 is depressurized by the stopper 34, it is not easily blocked. Further, even if the upper end side of the spare port 16 is opened by the stopper 34, the liquid does not immediately flow out of the first storage chamber 14. When accessing the first storage chamber 14 from the outside, an injection needle or the like that can penetrate the membrane of the stopper 34 is punctured. The posture of the punctured injection needle and the like is stabilized by the membrane of the stopper 34.
  • the spare port 16 is used when, for example, a drug or the like is injected into the cell suspension 40 stored in the first storage chamber 14.
  • the cell collection port 17 is provided at the lower end of the first storage chamber 14 in the vertical direction 53.
  • the lower end of the cell recovery port 17 is sealed and does not open to the outside.
  • the lower end side of the cell collection port 17 is cut and opened.
  • a cylindrical plug 35 is inserted into the cell recovery port 17 in a liquid-tight manner.
  • a cylindrical tube and a film that divides the inner space of the tube into one end side (upper side in FIG. 2) and the other end side (lower side in FIG. 2) are integrated with rubber or elastomer. It is formed.
  • the plug 35 does not easily block the cell collection port 17 even if it receives an external force or the like, or the first storage chamber 14 is decompressed.
  • the liquid does not immediately flow out of the first storage chamber 14.
  • an injection needle or the like that can penetrate the membrane of the plug 35 is punctured.
  • the posture of the punctured injection needle or the like is stabilized by the film of the stopper 35.
  • the cell collection port 17 is used when the cell suspension 40 stored in the first storage chamber 14 is caused to flow out.
  • a suspension hole 19 is provided between the cell injection port 15 and the reserve port 16 at the upper end of the first accommodating portion 11.
  • the suspension hole 19 is disposed near the center in the left-right direction 52 of the first housing portion 11. The suspension hole 19 is used when the first housing part 11 in a state where the second housing part 12 and the third housing part 13 are cut and separated is suspended and stored.
  • the second housing part 12 is connected to the first housing part 11 via the first connecting part 21.
  • the second housing part 12 is disposed on the right side in the left-right direction 52 of the first housing part 11 (right side in FIG. 1).
  • the second accommodating portion 12 has a second accommodating chamber 22 that is an internal space, and has a thin shape in which the length in the front-rear direction 51 is the shortest in comparison with the length in the left-right direction 52 and the vertical direction 53.
  • the second storage chamber 22 is a substantially rectangular parallelepiped space, the upper portion in the vertical direction 53 has the maximum length in the left-right direction 52, and the lower portion in the vertical direction 53 has a length in the left-right direction 52. It is curved into a funnel shape that gradually shortens downward.
  • the volume of the second storage chamber 22 is smaller than the volume of the first storage chamber 14, and is about 5 mL, for example.
  • the first connecting part 21 connects the first accommodating part 11 and the second accommodating part 12 at the upper part in the vertical direction 53.
  • the 1st connection part 21 is comprised by the two resin sheets which comprise the container 10 closely_contact
  • the length of the outer shape of the first connecting portion 21 in the front-rear direction 51 is shorter than the length in the up-down direction 53. That is, the first connecting portion 21 has a thin shape in the front-rear direction 51.
  • the first connecting portion 21 is provided with communication passages 23 and 24 extending in the left-right direction 52.
  • the communication passages 23 and 24 are spaced apart from each other in the vertical direction 53, and each communicates the first storage chamber 14 and the second storage chamber 22.
  • the communication passages 23 and 24 are disposed at positions offset upward from the center in the vertical direction 53 of the outer shape of the first storage portion 11 and the second storage portion 12. Further, the communication passages 23 and 24 are respectively opened at the ends in the left and right direction 52 of the position (upper portion) where the length in the left and right direction 52 is maximum in the first accommodation chamber 14 and the second accommodation chamber 22.
  • a sample collection port 25 is provided in the second storage chamber 22.
  • the sample collection port 25 is provided at the lower end in the up-down direction 53 of the second storage chamber 22.
  • the lower end of the sample collection port 25 is sealed and does not open to the outside.
  • the lower end side of the sample collection port 25 is cut and opened.
  • a cylindrical stopper 36 is inserted into the sample recovery port 25 in a liquid-tight manner. Since the structure of the plug 36 is the same as that of the plug 35, detailed description thereof is omitted here.
  • the sample collection port 25 is used when the cell suspension 40 stored in the second storage chamber 22 is caused to flow out.
  • the 3rd accommodating part 13 is connected with the 2nd accommodating part 12 through the 2nd connection part 26. As shown in FIG. The 3rd accommodating part 13 is arrange
  • the 3rd accommodating part 13 has the 3rd accommodating chamber 27 which is internal space, and is the thin shape with the length of the front-back direction 51 being the shortest compared with the length of the left-right direction 52 and the up-down direction 53. .
  • the third storage chamber 27 is a substantially rectangular parallelepiped space.
  • the volume of the 3rd storage chamber 27 is smaller than the volume of the 1st storage chamber 14 and the 2nd storage chamber 22, for example, is about 2 mL. Therefore, the sum (for example, about 7 mL) of the volume of the second storage chamber 22 and the volume of the third storage chamber 27 is smaller than the volume (for example, about 20 mL) of the first storage chamber 14.
  • the 2nd connection part 26 has connected the 2nd accommodating part 12 and the 3rd accommodating part 13 in the up-and-down direction 53 upper part.
  • the 2nd connection part 26 is comprised by the two resin sheets which comprise the container 10 closely_contact
  • the length of the outer shape of the second connecting portion 26 in the front-rear direction 51 is shorter than the length in the up-down direction 53. That is, the second connecting portion 26 has a thin shape in the front-rear direction 51.
  • the second connecting portion 26 is provided with communication passages 29 and 30 extending in the left-right direction 52.
  • the communication passages 29 and 30 are spaced apart from each other in the vertical direction 53, and each communicates the second storage chamber 22 and the third storage chamber 27.
  • the communication passages 29 and 30 are disposed at positions offset upward from the center in the vertical direction 53 of the outer shape of the second storage portion 12 and the third storage portion 13. Further, the communication passages 29 and 30 are respectively opened at the ends in the left and right direction 52 of the position (upper portion) where the length in the left and right direction 52 is maximum in the second accommodation chamber 22 and the third accommodation chamber 27.
  • a sample collection port 31 is provided in the third storage chamber 27.
  • the sample collection port 31 is provided at the lower end of the third storage chamber 27 in the vertical direction 53.
  • the lower end of the sample collection port 31 is sealed and does not open to the outside.
  • the lower end side of the sample collection port 31 is cut and opened.
  • a cylindrical stopper 37 is inserted into the sample recovery port 31 in a liquid-tight manner. Since the structure of the stopper 37 is the same as that of the stopper 35, detailed description thereof is omitted here.
  • the sample collection port 31 is used when the cell suspension 40 stored in the third storage chamber 27 flows out.
  • the sealing device 60 is an impulse heat sealer. As shown in FIG. 9, the sealing device 60 includes a first pressing portion 61 provided on a lower frame (not shown) and a second pressing portion 62 provided on an upper frame (not shown). Although not shown in detail in each drawing, the upper frame is provided so as to be able to contact and separate from the lower frame, and the first connecting part 21 or the second connecting part of the container 10 supported along the lower frame. The first connecting portion 21 or the second connecting portion 26 is sandwiched between the first pressing portion 61 and the second pressing portion 62 that are disposed so as to face the upper and lower directions 55. As a result, the communication passages 23 and 24 or the communication passages 29 and 30 are sealed by heat welding.
  • the first pressing portion 61 includes a first heater portion 63, a first protective cover 64, and a first transformer 65.
  • the first heater unit 63 is electrically connected to the first transformer 65 and is heated by electric power supplied from the first transformer 65.
  • the first protective cover 64 covers the upper side of the first heater unit 63.
  • the second pressing part 62 includes a second heater part 66, a second protective cover 67, and a second transformer 68.
  • the second heater unit 66 is electrically connected to the second transformer 68 and is heated by the electric power supplied from the second transformer 68.
  • the second protective cover 67 covers the lower side of the second heater portion 66.
  • the sealing apparatus 60 has the 1st press part 61 and the 2nd press part 62, the sealing apparatus which concerns on this invention as a sealing apparatus provided with only one of the 1st press part 61 or the 2nd press part 62 is provided. May be realized.
  • the first heater unit 63 extends in the left-right direction 57 out of the front-rear direction 56 and the left-right direction 57 orthogonal to the up-down direction 55.
  • the length along the left-right direction 57 of the 1st heater part 63 should just be sufficient to seal the communicating paths 23, 24, 29, and 30 of the container 10.
  • the first heater unit 63 includes a heater body 70 disposed in the center of the front-rear direction 56 and heat insulating materials 71, 72 disposed on both sides of the heater body 70 in the front-rear direction 56, respectively.
  • the heater body 70 is configured by a braid formed by assembling a plurality of thread-like nichrome wires, and the braid extends in the left-right direction 57 with a constant length along the front-rear direction 56.
  • the length along the front-rear direction 56 of the heater body 70 is sufficiently shorter than the length along the left-right direction 52 of the first connecting portion 21 and the second connecting portion 26 of the container 10, and the communication passages 23, 24, 29, It is long enough to maintain 30 seals.
  • the heater body 70 corresponds to a heat generating member.
  • the heat insulating materials 71 and 72 are disposed adjacent to the heater body 70.
  • a material of the heat insulating materials 71 and 72 for example, fluororesin (heat resistance, releasability), polytetrafluoroethylene, polyoxymethylene (POM), polypropylene (PP), polycarbonate (PC), polyamide resin, polyethylene terephthalate (PET).
  • the length of the heat insulating materials 71 and 72 along the left-right direction 57 is the same as the length of the heater body 70 along the left-right direction 57.
  • the length of the heat insulating materials 71 and 72 along the front-rear direction 56 is slightly longer than the length of the heater body 70 along the front-rear direction 56.
  • the length along the front-rear direction 56 of the heater body 70 and the heat insulating materials 71 and 72 as a whole is shorter than the length along the left-right direction 52 of the first connecting part 21 and the second connecting part 26 of the container 10.
  • the upper surfaces of the heat insulating materials 71 and 72 are substantially flush with the upper surface of the heater body 70. Therefore, when the container 10 comes into contact with the upper surface of the heater body 70, the container 10 also comes into contact with the upper surfaces of the heat insulating materials 71 and 72.
  • the heat insulating materials 71 and 72 correspond to a first heat insulating material and a second heat insulating material.
  • the 2nd heater part 66 is the structure by which the heat insulating material was arrange
  • the cell suspension 40 is filled into the container 10 and the first connecting part 21 and the second connecting part 26 are sealed, whereby a cell-filling container is manufactured.
  • the manufacturing method of a cell filling container is mainly classified into the following steps. (1) Until the first storage chamber 14 is filled with the cell suspension 40, the cell suspension 40 does not flow into the second storage chamber 22 through the communication passages 23 and 24, and the cells enter the first storage chamber 14. First step of filling the suspension 40. (2) The cell suspension 40 filled in the first storage chamber 14 is made to flow into the second storage chamber 22 through the communication passages 23 and 24, and the cell suspension filled in the first storage chamber 14 through the communication passages 29 and 30. A second step of allowing the turbid liquid 40 to flow into the third storage chamber 27; (3) In the second step, sealing the communication passages 23 and 24 and the communication passages 29 and 30 of the container 10 filled with the cell suspension 40.
  • Examples of cells filled in the container 10 include cells collected from living organisms such as umbilical cord blood and stem cells, but cells that can be cryopreserved using the container 10 are not limited to these.
  • the container 10 is maintained in a state in which the second storage chamber 22 and the third storage chamber 27 are located above the first storage chamber 14 in the gravity direction 54. Subsequently, the cell injection port 15 is connected to a decompression device such as an air pump or a syringe pump, and the first accommodation chamber 14 is decompressed by discharging air from the first accommodation chamber 14.
  • a decompression device such as an air pump or a syringe pump
  • the second storage chamber 22 and the third storage chamber 27 are in communication with the first storage chamber 14 through the communication passages 23, 24, 29, and 30, the second storage chamber 22 and the third storage chamber 27 are also decompressed. However, decompression of the second storage chamber 22 and the third storage chamber 27 is not necessarily required. Therefore, for example, only the first storage chamber 14 may be decompressed in a state where the communication passages 23 and 24 are closed by clips or the like.
  • the cell injection port 15 is disconnected from the decompression device, and the cell suspension 40 is injected into the first storage chamber 14 through the cell injection port 15 as shown in FIG. . Since the first storage chamber 14 is decompressed, the cell suspension 40 is filled into the first storage chamber 14 even if the first storage chamber 14 is not in communication with the atmosphere.
  • the second storage chamber 22 and the third storage chamber 27 are located above the first storage chamber 14 in the gravitational direction 54, and the communication paths 23 and 24 are opened on the uppermost side of the first storage chamber 14 in the gravitational direction 54. Therefore, the cell suspension 40 does not flow into the second storage chamber 22 and the third storage chamber 27 until the first storage chamber 14 is filled with the cell suspension 40. Accordingly, air 41 remains in the second storage chamber 22 and the third storage chamber 27. Note that after the first storage chamber 14 is filled with the cell suspension 40, the cell suspension 40 may flow from the first storage chamber 14 into the second storage chamber 22 and the third storage chamber 27.
  • the air 41 existing in the second storage chamber 22 and the third storage chamber 27 passes through the other of the communication passages 23 and 24 and the other of the communication passages 29 and 30. Thereby, the air 41 existing in the second storage chamber 22 and the third storage chamber 27 is replaced with the cell suspension 40 and moves to the first storage chamber 14.
  • the communication passages 23 and 24 and the communication passages 29 and 30 do not necessarily require the cell suspension 40 to circulate on one side and the air 41 to circulate on the other side.
  • the cell suspension 40 and the air 41 may flow simultaneously or alternately in the communication passages 23 and 24 and the communication passages 29 and 30.
  • a decompression device is connected to the cell injection port 15, The air 41 remaining in the first storage chamber 14 is extracted.
  • the container 10 is deformed so that the volume of the first storage chamber 14, the second storage chamber 22, and the third storage chamber 27 is bent by an amount corresponding to the remaining air 41.
  • the cell injection port 15 or the tube 18 is sealed by heat welding or the like. Thereby, the 1st storage chamber 14, the 2nd storage chamber 22, and the 3rd storage chamber 27 turn into a sealed space.
  • the communication paths 23 and 24 are sealed by heat welding in the first connection portion 21, and the communication paths 29 and 30 are sealed by heat welding in the second connection portion 26.
  • the These sealings are performed using a sealing device 60.
  • the thermal welding of the communication passages 23, 24 or the communication passages 29, 30 by the sealing device 60 that is, the sealing is performed by pressing the communication passages 23, 24 or the communication passages 29, 30 and closing them, and the communication passages 23, 24 or communication. And heat-welding the pressed portions of the passages 29 and 30.
  • the process of sealing the communication paths 23 and 24 and the process of sealing the communication paths 29 and 30 are the same, the detailed description will be made below with the process of sealing the communication paths 23 and 24 as an example.
  • the container 10 is positioned with respect to the sealing device 60 so that the communication paths 23 and 24 are aligned in the longitudinal direction of the first heater portion 63, that is, in the left-right direction 57.
  • the positioning of the container 10 may be realized by, for example, an operator holding the container 10 in his / her hand, or a jig for positioning the container 10 may be provided in the sealing device 60.
  • the positioning of the container 10 with respect to the sealing device 60 is performed so that the surfaces facing the front-rear direction 51 in each of the first housing portion 11, the second housing portion 12, and the third housing portion 13 are along the horizontal direction. It is preferable to support the container 10 with a jig or the like so that the passages 23 and 24 are along the horizontal direction.
  • the cell suspension 40 stored in the 1st accommodating part 11, the 2nd accommodating part 12, and the 3rd accommodating part 13 moves through the communicating paths 23 and 24, respectively.
  • the first storage unit 11 is positioned below the second storage unit 12 and the third storage unit 13, the cell suspension stored in the second storage unit 12 and the third storage unit 13 by gravity.
  • the originally planned amount is obtained when the turbid liquid 40 flows into the first housing portion 11 and the inner space of the first housing portion 11 expands so that the wall defining the inner space of the first housing portion 11 bends outward.
  • a larger amount of cell suspension 40 is stored in the first storage unit 11, and a smaller amount of cell suspension 40 than the originally planned amount is stored in the second storage unit 12 and the third storage unit 13.
  • the surfaces facing the front-rear direction 51 in each of the first housing portion 11, the second housing portion 12, and the third housing portion 13 are the first connecting portion 21 and the second connecting portion 26. Is sandwiched between a pair of support members having a flat plate shape without interfering with the first heater unit 63 and the second heater unit 66 without interfering with the first heater unit 63 and the second heater unit 66. Preferably it is held.
  • the communication passages 23 and 24 are sandwiched between the heat insulating materials 71 and 72 of the first heater portion 63 and the heat insulating material (not shown) of the second heater portion 66, thereby communicating with each other.
  • the cell suspension 40 is pushed out from a part 43 of the passages 23, 24, but the first storage part 11, the second storage part 12, and the third storage part 13 are sandwiched and held by a pair of flat support members. As a result, any one of the first accommodating portion 11, the second accommodating portion 12, and the third accommodating portion 13 is suppressed from being greatly bent. If the first storage part 11, the second storage part 12, and the third storage part 13 are not sandwiched between the pair of support members, the cell suspension 40 is introduced from a part 43 of the communication passages 23, 24.
  • any one of the first accommodation portion 11, the second accommodation portion 12, and the third accommodation portion 13 is easily bent more than the other accommodation portions, and as a result, the pushed cell suspension
  • the turbid liquid 40 may be biased to flow into any of the storage portions that are largely bent
  • the first storage portion 11, the second storage portion 12, and the third storage portion 13 are a pair of support members having a flat plate shape. The occurrence of such a problem is suppressed by being sandwiched by, for example.
  • the first storage portion 11 and the second storage portion 12 of the container 10 positioned as described above are located at opposite positions in the front-rear direction 56 of the first heater portion 63, respectively.
  • the second heater portion 66 is not shown in FIG. 11, the second heater portion 66 is positioned to face the first heater portion 63 in the up-down direction 55 (direction perpendicular to the paper surface of FIG. 11). Yes.
  • the second heater portion 66 is lowered and contacts the first heater portion 63.
  • the communication paths 23 and 24 of the container 10 are sandwiched and pressed between the heat insulating materials 71 and 72 of the first heater section 63 and the heat insulating material (not shown) of the second heater section 66, and the communication path 23. 24 are crushed and closed.
  • the communication paths 23 and 24 are sandwiched between the heat insulating materials 71 and 72 of the first heater unit 63 and the heat insulating material (not shown) of the second heater unit 66.
  • the cell suspension 40 moves from a part 43 of the communicating passages 23 and 24 to the first storage unit 11 side or the second storage unit 12 side.
  • the first transformer 65 and the heater body 70 (not shown) of the first heater unit 63 and the heater body (not shown) of the second heater unit 66 are connected. Electric power is supplied from each second transformer 68.
  • the heater main body 70 of the first heater section 63 and the heater main body (not shown) of the second heater section 66 generate heat, and the heater main body of the first heater section 63, that is, approximately the center of the part 43 in the front-rear direction 56.
  • the portions of the 70 and the second heater portion 66 that are in contact with the heater main body (not shown) are thermally welded to form the sealed portion 42.
  • the communication passages 23 and 24 are provided in the first passage. After being sandwiched between the heat insulating materials 71 and 72 of the first heater portion 63 and the heat insulating material (not shown) of the second heater portion 66, the heater main body 70 of the first heater portion 63 and the heater main body of the second heater portion 66 A time lag of at least about 0.1 to 1.0 seconds is preferably set until power is supplied to (not shown).
  • the time lag described above includes, for example, an operation for sandwiching the communication paths 23 and 24 between the heat insulating materials 71 and 72 of the first heater unit 63 and a heat insulating material (not shown) of the second heater unit 66, and the first If the operation of supplying power to the heater main body 70 of the heater unit 63 and the heater main body (not shown) of the second heater unit 66 (for example, switch-on) is independent, the sandwiching is completed. This is realized by the control that disables the switch-on for supplying power until the above-described time lag elapses after the detection.
  • an operation for sandwiching the communication passages 23 and 24 between the heat insulating materials 71 and 72 of the first heater section 63 and a heat insulating material (not shown) of the second heater section 66 is, for example, in a direction to bring the first heater unit 63 into contact with the second heater unit. If it is a so-called one-action configuration that is completed by a series of moving operations, it is realized by setting the above-mentioned time lag from when switch-on for power supply is input to when power is actually supplied Is done.
  • the communication paths 29 and 30 are also heat-sealed by the sealing apparatus 60 in the 2nd connection part 26, and are sealed.
  • Each of the first storage chamber 14, the second storage chamber 22, and the third storage chamber 27 becomes a sealed space by the sealing portion 42. Thereafter, as shown in FIG. 8, in the first connecting portion 21 and the second connecting portion 26, the communication passages 23 and 24 or the communication passages 29 and 30 are cut along the sealing portions 42 that are heat-welded. Thus, the first housing portion 11, the second housing portion 12, and the third housing portion 13 are separated. In addition, if the 2nd accommodating part 12 and the 3rd accommodating part 13 are stored in the same place, the sealing part 42 of the 2nd connection part 26 will not be cut
  • the cell collection port 17 or the sample is used.
  • the collection ports 25 and 31 are opened, and the cell suspension 40 flows out from the first storage chamber 14, the second storage chamber 22, or the third storage chamber 27.
  • the cell suspension 40 is preferentially filled in the first storage chamber 14 in the first step using gravity, and the second storage chamber 22 and the third storage are loaded from the first storage chamber 14 in the second step. Since the cell suspension 40 can be caused to flow into the chamber 27, each process is easily realized.
  • the first storage chamber 14 since the first storage chamber 14 is decompressed and then filled with the cell suspension 40, it is not necessary to connect the first storage chamber 14 to the atmosphere.
  • the air 41 remaining in the first storage chamber 14 is removed, so that the cells in the first storage chamber 14 are removed. It is possible to prevent the suspension 40 from being affected by the atmosphere.
  • the first storage chamber 14 is sealed by sealing the communication passages 23, 24, 29, and 30.
  • Each of the second storage chamber 22 and the third storage chamber 27 can be a sealed space.
  • the second storage chamber 22 is easily filled with the cell suspension 40 filled in the first storage chamber 14.
  • the communication passages 23, 24, 29, and 30 are pressed and closed, and the pressed portions are sealed by thermal welding. There is no 40. Therefore, heat at the time of heat welding is not easily transmitted to the cell suspension 40. Further, since the heater main body 70 is heated and thermally welded while the communication passages 23, 24, 29, and 30 are pressed and closed by the sealing device 60 heat insulating materials 71 and 72, cells are placed at the positions where heat welding is performed. The suspension 40 does not exist, and heat at the time of heat welding is difficult to transfer to the cell suspension 40.
  • the container 10 since the container 10 includes the second storage portion 12 and the third storage portion 13, a plurality of test samples can be obtained.
  • the second storage chamber 22 and the volume of the third storage chamber 27 are the first storage chamber. It becomes easy to be filled with the liquid with which 14 was filled.
  • the lengths of the outer shapes of the first connecting portion 21 and the second connecting portion 26 in the front-rear direction 51 are shorter than the length in the up-down direction 53, and the communication paths 23 and 24 are formed in the first receiving portion 11 and the second receiving portion 12.
  • the communication passages 29 and 30 are located above the center of the outer shape of the second housing part 12 and the third housing part 13 in the up-down direction 53. Therefore, the communication passages 23, 24, 29, and 30 can be easily sealed by heat welding or the like.
  • the communication passages 23 and 24 are respectively opened at the ends in the left and right direction 52 of the position (upper portion) where the length in the left and right direction 52 is the maximum in the first storage chamber 14 and the second storage chamber 22. Since each of the communication passages 29 and 30 is open at the end in the left and right direction 52 of the position (upper portion) where the length in the left and right direction 52 is the maximum in the second accommodation chamber 22 and the third accommodation chamber 27, When the first accommodating portion 11 is on the upper side and the third accommodating portion 13 is on the lower side, air can easily escape from the second accommodating chamber 22 and the third accommodating chamber 27.
  • the second storage chamber 22 and the third storage chamber 27 are positioned above the first storage chamber 14 in the gravitational direction 54 so that the first storage chamber 14 is a cell.
  • the cell suspension 40 is prevented from flowing into the second storage chamber 22 and the third storage chamber 27 until the suspension 40 is filled with the suspension 40.
  • the communication paths 23 and 24 are clipped without using gravity. So that the cell suspension 40 does not flow into the second storage chamber 22 and the third storage chamber 27 until the first storage chamber 14 is filled with the cell suspension 40. Also good.
  • the second storage chamber 22 and the third storage chamber 27 are positioned below the first storage chamber 14 in the direction of gravity 54, so that the cell suspension filled in the first storage chamber 14 is obtained.
  • the liquid 40 flows into the second storage chamber 22 and the third storage chamber 27, but without using the gravity, for example, by pressing the first storage portion 11 from the outside, the second storage chamber 14 can store the second storage chamber.
  • the cell suspension 40 may flow into the chamber 22 and the third storage chamber 27.
  • the cell suspension 40 as a test sample is stored in the second storage chamber 22 and the third storage chamber 27. If a plurality of test samples are not necessary, the third storage portion 13 is provided. Instead, only the second container 12 may be provided in the container 10 for storing the test sample. On the other hand, if three or more test samples are required, in addition to the second storage unit 12 and the third storage unit 13, a similar storage unit may be provided in the container 10 for storing the test sample.
  • the position in the first storage chamber 14, the second storage chamber 22, and the third storage chamber 27 where the length in the left-right direction 52 is maximum is the upper portion of the vertical direction 53.
  • the position where the length of the uppermost portion may be the maximum may be the lower portion of the vertical direction 53 or the central portion.
  • the communication paths 23, 24, 29 and 30 are arranged in a biased manner in the lower portions of the first housing part 11, the second housing part 12 and the third housing part 13.
  • cell (sample) recovery ports are provided below the first storage unit 11, the second storage unit 12, and the third storage unit 13. The positions of these ports are appropriately changed. Also good.
  • the sealing device 60 separately seals the communication paths 23 and 24 of the first connection part 21 and the communication paths 29 and 30 of the second connection part 26, but the first pressing part 61 and the second pressing part 62.
  • the sealing device 60 may be configured such that two sets are provided and the communication paths 23 and 24 of the first connection part 21 and the communication paths 29 and 30 of the second connection part 26 can be sealed simultaneously.
  • the communication passages 23 and 24 of the first connection portion 21 of the container 10 and the communication passages 29 and 30 of the second connection portion 26 may be sealed by a device other than the sealing device 60.

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Abstract

L'invention vise à procurer un procédé de fabrication d'un récipient pour le remplissage de cellules, approprié pour stocker des échantillons à tester d'un fluide contenant des cellules. L'invention concerne un récipient (10) pourvu : d'une première unité de réception (11) divisant une première chambre de réception (14) ; d'une deuxième unité de réception (12) reliée à la première unité de réception (11) par l'intermédiaire d'une première unité de liaison (21) et divisant une deuxième chambre de réception (22) ; et des trajets de liaison (23, 24) ménagés dans la première unité de liaison (21) pour relier la première chambre de réception (14) et la deuxième chambre de réception (22). Dans un état où le fluide de suspension des cellules (40) ne s'écoule pas vers la deuxième chambre de réception (22) par les trajets de liaison (23, 24) jusqu'à ce que la première chambre de réception (14) soit remplie d'un fluide de suspension des cellules (40), la première chambre de réception (14) est remplie avec le fluide de suspension des cellules (40), et ensuite, le fluide de suspension des cellules (40), qui a rempli la première chambre de réception (14), s'écoule dans la deuxième chambre de réception (22) par les trajets de liaison (23, 24).
PCT/JP2015/059189 2014-03-28 2015-03-25 Procédé de fabrication d'un récipient de remplissage de cellules, récipient pour remplissage de cellules et dispositif d'étanchéité WO2015147077A1 (fr)

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JP2020103142A (ja) * 2018-12-27 2020-07-09 大日本印刷株式会社 サンプリング機能付き容器
JP2021019552A (ja) * 2019-07-29 2021-02-18 株式会社カネカ 細胞充填システム及び細胞医薬製剤の製造方法
JP2022119803A (ja) * 2017-10-09 2022-08-17 テルモ ビーシーティー バイオテクノロジーズ，エルエルシー マルチパート凍結乾燥容器

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