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WO2018123619A1 - Sac équipé d'un filtre pour cellules et procédé de production correspondant - Google Patents

Sac équipé d'un filtre pour cellules et procédé de production correspondant Download PDF

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Publication number
WO2018123619A1
WO2018123619A1 PCT/JP2017/044925 JP2017044925W WO2018123619A1 WO 2018123619 A1 WO2018123619 A1 WO 2018123619A1 JP 2017044925 W JP2017044925 W JP 2017044925W WO 2018123619 A1 WO2018123619 A1 WO 2018123619A1
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WO
WIPO (PCT)
Prior art keywords
mesh
filter
filter member
polyethylene
container body
Prior art date
Application number
PCT/JP2017/044925
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English (en)
Japanese (ja)
Inventor
郷史 田中
串田　尚
あゆみ 石割
安達　慎太郎
Original Assignee
東洋製罐グループホールディングス株式会社
帝人株式会社
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.)
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Publication date
Application filed by 東洋製罐グループホールディングス株式会社, 帝人株式会社 filed Critical 東洋製罐グループホールディングス株式会社
Publication of WO2018123619A1 publication Critical patent/WO2018123619A1/fr

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    • 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
    • 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
    • C12M3/06—Tissue, human, animal or plant cell, or virus culture apparatus with filtration, ultrafiltration, inverse osmosis or dialysis means

Definitions

  • the present invention relates to a cell bag with a filter for easily and efficiently performing various operations such as cell and medium fractionation, filtration, and washing performed during cell culture, and a method for producing the same.
  • the present applicant has repeatedly studied a cell culture system capable of performing cell culture efficiently while constructing a closed system environment to reduce the risk of contamination.
  • Patent Document 1 a culture container for culturing cells, a medium storage container for storing a medium and the like, a cell injection container for injecting cells, and a cell suspension after culture
  • a cell culture kit is proposed in which a cell collection container to be collected is connected by a conduit to construct a closed system environment. According to such a cell culture kit, it is possible to carry out from cell injection to medium addition, sampling, and collection while maintaining a closed system in the kit.
  • Patent Document 1 in recovering cultured cells, the culture vessel is allowed to stand to settle the cells in the cell suspension, and then the supernatant of the cell suspension is discharged, An example is shown in which the concentrated cell suspension is transferred from the culture vessel to the cell collection vessel after being reduced.
  • cell culture usually takes a period of several days to several weeks, so that cell growth is not inhibited by the depletion of medium components or accumulation of cell metabolites,
  • the medium may be changed as necessary.
  • the cells in the cell suspension must be allowed to settle by allowing the culture vessel to stand before draining the cell suspension supernatant. It takes time to settle the cells.
  • cells may be mixed into the supernatant by the discharge operation, and the cells may be discharged together with the supernatant.
  • it is desirable to remove as much of the old medium as possible, and to add more new medium.
  • cell contamination is avoided. It will not be possible.
  • the flow path diameter of the port is usually about 1 to 10 mm, and the area of the filter is relatively small. Therefore, the trapped cells are likely to be clogged, and the old medium may be prevented from being discharged. is there. Furthermore, there is a possibility that the cells captured by the filter do not return to the culture container and die while being captured by the filter.
  • the present applicant partitions the inside of the container body with a filter member, so that, for example, when exchanging the medium, the cells in culture remain in one partition. In view of this, only the old medium can be discharged from the other compartment, and various studies have been made to realize this.
  • the present invention provides a filter for partitioning the inside of a container body with a filter member in order to easily and efficiently perform various operations such as cell and medium fractionation, filtration, and washing performed during cell culture.
  • An object of the present invention is to provide a cell bag with a filter whose seal strength with a member is increased, and a method for producing the same.
  • the cell bag with a filter includes a container main body and an injecting / extracting port, the inside of the container main body is partitioned by a filter member, and an injecting / extracting port is provided in at least one section.
  • the filter member comprises a main mesh made of polyester or polyamide having a mesh opening of 20 to 300 ⁇ m, and a polyethylene sub-mesh having a mesh opening of 100 to 4000 ⁇ m arranged on both front and back surfaces thereof, and the container body has at least Between the flexible film containing a polyethylene layer, the said polyethylene layer is made to oppose, and the peripheral part is heat-sealed on both sides of the said filter member, and the heat-fusion part formed in the peripheral part of the said container main body In the above, the polyethylene sub-mesh is melted and fused to each other while entering the gap of the main mesh Both are configured to be fused to the polyethylene layer of the flexible film.
  • the method for producing a cell bag with a filter according to the present invention comprises a container main body and an injection port, and the inside of the container main body is partitioned by a filter member, and the injection port is provided in at least one partition.
  • the method of manufacturing a cell bag with a filter provided with a sub mesh made of polyethylene having an opening of 100 to 4000 ⁇ m is provided on both sides of a main mesh made of polyester or polyamide having an opening of 20 to 300 ⁇ m.
  • the filter member is prepared, the polyethylene layer is opposed between at least a flexible film including a polyethylene layer, the filter member is sandwiched, and the peripheral portions thereof are heat-sealed.
  • the present invention it is possible to provide a cell bag with a filter in which the sealing strength between the flexible film forming the container body and the filter member is increased and the bag breaking strength is improved.
  • FIG. 1 is an explanatory view showing an example of a cell bag with a filter according to the present embodiment
  • FIG. 1 (a) is a plan view of the cell bag with filter 1
  • FIG. 1 (b) is a filter.
  • FIG. 1C is a cross-sectional view taken along the line AA of FIG.
  • a cell bag with filter 1 shown in FIG. 1 includes a container body 2 and injection ports 3a and 3b. And the inside of the container main body 2 is divided into two chambers by the filter member 4, and injection ports 3a and 3b are provided in the respective compartments (first compartment 1st and second compartment 2nd).
  • the injection ports 3a and 3b are portions that serve as entrances and exits when injecting and discharging culture media and cells.
  • the injection ports 3a and 3b are provided by attaching a tubular member through which the culture media or cells can flow to the container body 2.
  • the tubular members forming the injection ports 3a and 3b are formed into a predetermined shape by injection molding, extrusion molding, or the like using a thermoplastic resin such as polyethylene, polypropylene, vinyl chloride, polystyrene elastomer, FEP, etc. Can be molded.
  • the filter member 4 is configured using two types of meshes having different mesh sizes, and includes a main mesh 4a having a relatively small mesh size and a sub-mesh 4b having a relatively large mesh size disposed on both front and back surfaces. It has become.
  • a polyester mesh woven from a polyester resin fiber such as polyethylene terephthalate or a polyamide mesh woven from a polyamide resin fiber such as nylon is used.
  • a polyethylene mesh formed by weaving polyethylene resin fibers is used for the sub-mesh 4b.
  • the polyethylene-type resin fiber which forms the submesh 4b consists of the same kind of polyethylene-type resin as the polyethylene layer 20a of the flexible film 20 mentioned later.
  • the mesh size of these meshes 4a and 4b can be appropriately selected according to the usage mode of the filter-equipped cell bag 1.
  • the main mesh 4a has an opening of 20 to 300 ⁇ m
  • the submesh 4b has a mesh size of 20 to 300 ⁇ m.
  • These mesh sizes are selected in the range of 100 to 4000 ⁇ m. At this time, it is preferable to select from the above range so that the opening of the sub-mesh 4b is 2 to 10 times larger than the opening of the main mesh 4a.
  • the usage example of the cell bag 1 with a filter is mentioned later.
  • the container body 2 is formed by making the flexible film 20 into a bag shape.
  • a flexible film 20 including at least a polyethylene layer 20 a is used as the flexible film 20 forming the container body 2.
  • a flexible film 20 includes, for example, polypropylene, ethylene-vinyl acetate copolymer, polyester, polyamide, silicone elastomer, polystyrene elastomer, tetrafluoroethylene, in addition to a polyethylene resin film including a polyethylene layer 20a as a single layer.
  • the -It may be a laminated film in which the polyethylene layer 20a is laminated on one or more layers made of a thermoplastic resin such as hexafluoropropylene copolymer (FEP) so that the polyethylene layer 20a is located on the surface layer.
  • FEP hexafluoropropylene copolymer
  • the flexible film 20 which forms the container main body 2 has transparency to such an extent that the progress of cell culture, the state of internal cells, the color of the medium, and the like can be confirmed.
  • the container body 2 has two flexible films 20 cut to a desired size, and the two layers of the flexible films 20 are aligned and overlapped, with the polyethylene layer 20a facing each other between the two flexible films 20. Then, these peripheral portions can be formed by heat-sealing with the filter member 4 sandwiched therebetween.
  • the peripheral part of the container body 2 What is necessary is just to heat-seal
  • FIG. 2 is an explanatory diagram schematically showing a production example of the cell bag 1 with a filter, and shows the arrangement of the flexible film 20, the filter member 4, and the injection ports 3a and 3b.
  • the sub-mesh 4b (specifically, the sub-mesh) disposed on both the front and back surfaces of the main mesh 4a in the heat-sealing portion 5 formed at the peripheral portion thereof. 4b forming polyethylene-based resin fibers) are melted and are fused to each other while penetrating into the gaps of the main mesh 4a, and are also fused to the polyethylene layer 20a of the flexible film 20. As a result, the sealing strength between the flexible film 20 forming the container body 2 and the filter member 4 is increased, and the bag breaking strength of the cell bag with filter 1 is improved.
  • the filter member 4 In the state in which the filter member 4 is sandwiched between the flexible films 20, these peripheral portions are heat-sealed to form the container body 2, and the filter member 4 is also cut into a desired size.
  • melting part 5 it is preferable to align so that the edge of the main mesh 4a may be located inside a container rather than the edge of the submesh 4b. Furthermore, it is more preferable to trim the sub-mesh 4b so that the edge of the sub-mesh 4b is positioned inside the container rather than the edge of the flexible film 20 that forms the container body 2. 3 corresponds to the BB cross section of FIG.
  • An end edge of the flexible film 20 is indicated by a one-dot chain line ⁇
  • an end edge of the main mesh 4a is indicated by a one-dot chain line ⁇
  • an end edge of the sub-mesh 4b is indicated by a one-dot chain line ⁇ .
  • the polyethylene layers 20a of the flexible film 20 are melted and fused directly or via the sub-mesh 4b. .
  • the outermost edge portion of the container body 2 is more strongly heat-sealed, and the bag breaking strength of the cell bag with filter 1 is further improved.
  • FIG. 4 shows an example in which the filter-equipped cell bag 1 shown in FIG. 1 is used as a culture bag in which operations such as medium replacement can be performed simply and efficiently.
  • the cells C to be cultured are injected into the container body 2 filled with the medium from the injection port 3b provided in the second section 2nd, and the culture is performed.
  • a main mesh 4a of the filter member 4 having a mesh size that does not allow the cell C to pass through is selected.
  • the mesh size of the submesh 4b of the filter member 4 should just be larger than the mesh size of the main mesh 4a, and can be suitably selected irrespective of the magnitude
  • the old medium When exchanging the medium during the culture period, the old medium is discharged from the inlet / outlet port 3a provided in the first section 1st (see FIG. 4B). At this time, since the cells C being cultured can remain in the second compartment 2nd, it can be prevented from being discharged together with the old medium, and more of the old medium can be discharged. After discharging the old medium, the same amount of new medium is injected from the injection port 3a provided in the first section 1st (see FIG. 4C), and the culture is continued.
  • the medium exchange during the culture period in particular, the operation of discharging the old medium can be performed easily and efficiently.
  • the cell bag with filter 1 in this use example discharges the content liquid in the container from the injection port 3a provided in the first section 1st in a state where the cells C remain in the second section 2nd. Therefore, for example, when used as a culture container for the cell culture kit of Patent Document 1, the cell recovery operation exemplified in Patent Document 1 is also simple and efficient without taking time for cell sedimentation. Can be done well.
  • the washing / recovery mechanism can be introduced into the cell culture kit of Patent Document 1 by repeating the injection and discharge of the washing liquid to perform the washing operation of the cells.
  • a mesh size that does not allow passage of the cells C to be cultured is selected, and the cells are placed in one partition 2nd partitioned by the filter member 4.
  • a medium, a washing solution, and the like are supplied from the injection port 3a provided in the other section 1st partitioned by the filter member 4. Can be injected.
  • This use example is one use example of the cell bag 1 with a filter according to the present embodiment, and the cells performed during cell culture by using the bag 1 with a filter cell according to the present embodiment for cell culture.
  • the various operations such as fractionation, filtration, washing, and filtration of the medium can be carried out simply and efficiently.
  • the inside of the container body 2 is partitioned into two chambers
  • a plurality of similar filter members 4 may be used to partition into three or more chambers.
  • the pouring port may be provided in at least one section, and there may be a section in which the pouring port is not provided.
  • the present invention can be used as a technique for efficiently culturing cells in the fields of pharmaceutical science and biochemistry.

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

Abstract

Selon l'invention, un corps 2 de récipient est formé par la prise en sandwich, entre des films souples 20 dont chacun comprend au moins une couche de polyéthylène 20a et qui sont disposés de manière à ce que les couches de polyéthylène 20a respectives sont orientées l'une vers l'autre, un élément de filtre 4 comprenant : une toile 4a principale en polyester ou en polyamide présentant des ouvertures de 20-300 µm ; et des sous-toiles 4b en polyéthylène qui présentent des ouvertures de 100-4000 μm et qui sont disposées sur les surfaces avant et arrière de la toile principale, puis par thermoscellage des films souples à la périphérie correspondante. Dans une partie thermoscellée 5 formée à la périphérie du corps 2 de récipient, les sous-toiles 4b sont fondues de manière à être liées par fusion les unes aux autres tout en infiltrant les vides dans la toile principale 4a et à être liées par fusion aux couches de polyéthylène 20a des films souples 20. Cette configuration permet d'améliorer la force d'étanchéité avec l'élément de filtre lorsque l'intérieur du corps de récipient doit être divisé par l'élément de filtre de façon à permettre une réalisation simple et efficace de diverses opérations nécessaires pour effectuer une culture cellulaire, telle qu'un fractionnement, une filtration et un lavage de cellules.
PCT/JP2017/044925 2016-12-26 2017-12-14 Sac équipé d'un filtre pour cellules et procédé de production correspondant WO2018123619A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2016250528A JP6803221B2 (ja) 2016-12-26 2016-12-26 フィルタ付き細胞用バッグ、及びその製造方法
JP2016-250528 2016-12-26

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WO2018123619A1 true WO2018123619A1 (fr) 2018-07-05

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TW (1) TW201829766A (fr)
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020085301A1 (fr) * 2018-10-23 2020-04-30 帝人株式会社 Filtre pour filtrage, récipient avec filtre, et procédé d'élimination de matière étrangère dans une suspension cellulaire
CN112996901A (zh) * 2018-09-11 2021-06-18 日产化学株式会社 分离装置及使用该分离装置对分离对象物进行分离的方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110189772A1 (en) * 2008-05-26 2011-08-04 Marc Benbunan Bioreactor with dual compartment
WO2011142667A1 (fr) * 2010-05-12 2011-11-17 Xpand Biotechnology B.V. Poche de culture cellulaire
CN205710773U (zh) * 2016-04-12 2016-11-23 福建三一造血技术有限公司 高效细胞培养袋

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110189772A1 (en) * 2008-05-26 2011-08-04 Marc Benbunan Bioreactor with dual compartment
WO2011142667A1 (fr) * 2010-05-12 2011-11-17 Xpand Biotechnology B.V. Poche de culture cellulaire
CN205710773U (zh) * 2016-04-12 2016-11-23 福建三一造血技术有限公司 高效细胞培养袋

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112996901A (zh) * 2018-09-11 2021-06-18 日产化学株式会社 分离装置及使用该分离装置对分离对象物进行分离的方法
WO2020085301A1 (fr) * 2018-10-23 2020-04-30 帝人株式会社 Filtre pour filtrage, récipient avec filtre, et procédé d'élimination de matière étrangère dans une suspension cellulaire
CN113164844A (zh) * 2018-10-23 2021-07-23 帝人株式会社 过滤用过滤器、带过滤器的容器和细胞悬浮液中的异物去除方法
JPWO2020085301A1 (ja) * 2018-10-23 2021-12-16 帝人株式会社 ろ過用フィルター、フィルター付容器、及び細胞懸濁液中の異物除去方法
EP3871748A4 (fr) * 2018-10-23 2022-08-03 Teijin Limited Filtre pour filtrage, récipient avec filtre, et procédé d'élimination de matière étrangère dans une suspension cellulaire
CN113164844B (zh) * 2018-10-23 2022-11-22 Jcr制药股份有限公司 过滤用过滤器、带过滤器的容器和细胞悬浮液中的异物去除方法
JP7291152B2 (ja) 2018-10-23 2023-06-14 Ｊｃｒファーマ株式会社 ろ過用フィルター、フィルター付容器、及び細胞懸濁液中の異物除去方法

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JP2018102169A (ja) 2018-07-05
JP6803221B2 (ja) 2020-12-23
TW201829766A (zh) 2018-08-16

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