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WO2018123619A1 - Filter-equipped bag for cells and production method therefor - Google Patents

Filter-equipped bag for cells and production method therefor 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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WIPO (PCT)
Prior art keywords
mesh
filter
filter member
polyethylene
container body
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PCT/JP2017/044925
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French (fr)
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
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Publication of WO2018123619A1 publication Critical patent/WO2018123619A1/en

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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

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  • 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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Abstract

A container body 2 is formed by sandwiching, between flexible films 20 each of which includes at least a polyethylene layer 20a and which are disposed so as to have the respective polyethylene layers 20a facing each other, a filter member 4 comprising: a polyester or polyamide main mesh 4a having openings of 20-300 μm; and polyethylene submeshes 4b which have openings of 100-4000 μm and which are disposed on the front and back surfaces of the main mesh, and then by heat-sealing the flexible films at the periphery thereof. In a heat-sealed part 5 formed at the periphery of the container body 2, the submeshes 4b are melted so as to be fusion-bonded to each other while infiltrating voids in the main mesh 4a, and to be fusion-bonded with the polyethylene layers 20a of the flexible films 20. This configuration is able to enhance the sealing strength with the filter member when the interior of the container body is to be partitioned by the filter member so as to allow for a simple and efficient way of performing various operations necessary to carry out cell culturing, such as fractionation, filtration, and washing of cells.

Description

フィルタ付き細胞用バッグ、及びその製造方法Cell bag with filter and method for producing the same
　本発明は、細胞培養に際して行われる細胞や培地の分画、ろ過、洗浄などの種々の操作を簡便、かつ、効率良く行うためのフィルタ付き細胞用バッグ、及びその製造方法に関する。 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.
　近年、医薬品の生産、遺伝子治療、再生医療、免疫療法などの医薬学・生化学分野において、細胞（組織、微生物、ウイルスなどを含む）を人工的な環境下で効率良く大量に培養することが求められている。 In recent years, in the fields of pharmacology and biochemistry such as pharmaceutical production, gene therapy, regenerative medicine, and immunotherapy, cells (including tissues, microorganisms, viruses, etc.) can be efficiently cultured in large quantities in an artificial environment. It has been demanded.
　このような要求に応えるべく、本出願人は、閉鎖系の環境を構築してコンタミネーションのリスクを低減しつつ、効率的に細胞培養を行うことができる細胞培養システムについて検討を重ねてきた。 In order to meet such demands, 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.
　例えば、特許文献１において、細胞を培養するための培養容器と、培地等を貯蔵しておくための培地貯蔵容器と、細胞を注入するための細胞注入容器と、培養後の細胞懸濁液を回収する細胞回収容器とを導管によって連結し、閉鎖系の環境を構築してなる細胞培養用キットを提案している。このような細胞培養用キットによれば、細胞注入から培地追加、サンプリング、回収までをキット内で閉鎖系を維持しながら行うことが可能となる。 For example, in 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.
国際公開２０１３／１１４８４５号パンフレットInternational Publication 2013/114845 Pamphlet
　ところで、特許文献１には、培養された細胞を回収するにあたり、培養容器を静置して細胞懸濁液中の細胞を沈降させた後に、細胞懸濁液の上澄みを排出し、液量を低減させてから、濃縮された細胞懸濁液を培養容器から細胞回収容器に移送する例が示されている。 By the way, in 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.
　また、細胞の培養には、通常、数日～数週間の期間を要するため、培地成分の枯渇や細胞の代謝物の蓄積によって細胞の成長が阻害されてしまわないように、培養期間中に、必要に応じて培地交換を行うことがある。培地交換を行うに際しては、細胞懸濁液の上澄みを排出することによって古い培地を取り除くとともに、それに代えて新しい培地を追加することが考えられる。 In addition, 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. When exchanging the medium, it is conceivable to remove the old medium by discharging the supernatant of the cell suspension and add a new medium instead.
　しかしながら、このようにして細胞の回収や培地交換を行うには、細胞懸濁液の上澄みを排出するに先立って、培養容器を静置して細胞懸濁液中の細胞を沈降させておかなければならず、細胞を沈降させるまでに時間を要してしまう。一方、十分な時間をかけて細胞懸濁液中の細胞を沈降させても、排出操作によって上澄みに細胞が混入し、上澄みと一緒に細胞が排出されてしまう虞もある。特に、培地交換を行うにあたっては、古い培地ができるだけ多く取り除かれるようにして、新しい培地がより多く追加されるようにすることが望まれるが、上澄みの排出量が多くなるほど、細胞の混入が避けられなくなってしまう。 However, in order to recover cells and change medium in this way, 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. On the other hand, even if the cells in the cell suspension are allowed to settle for a sufficient time, cells may be mixed into the supernatant by the discharge operation, and the cells may be discharged together with the supernatant. In particular, when exchanging the medium, it is desirable to remove as much of the old medium as possible, and to add more new medium. However, as the amount of supernatant discharged increases, cell contamination is avoided. It will not be possible.
　また、培地交換を行う際に、古い培地と一緒に細胞が排出されてしまうのを防ぐには、培養容器に設けられたポートの流路にフィルタを取り付けることも考えられる。
　しかしながら、ポートの流路径は、通常、１～１０ｍｍ程度であり、フィルタの面積も比較的小さくなるため、捕捉された細胞によってフィルタに目詰まりが生じ易く、古い培地の排出を妨げてしまう虞がある。さらに、フィルタに捕捉された細胞が培養容器内に戻らずに、フィルタに捕捉されたまま死滅してしまう虞もある。 In order to prevent the cells from being discharged together with the old medium when exchanging the medium, it is conceivable to attach a filter to the flow path of the port provided in the culture container.
However, 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.
　このような背景技術に鑑みて、本出願人は、容器本体の内部をフィルタ部材によって区画することで、例えば、培地交換を行う際には、培養中の細胞を一方の区画に留まらせておいて、他方の区画から古い培地のみを排出させることができると考え、これを実現するために種々の検討を重ねてきた。 In view of such background art, 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.
　しかしながら、フィルタ部材として用いるには、細胞の通過を許容しない程度にメッシュサイズの小さいものが容易に入手できるポリエステル製又はポリアミド製のメッシュが好適であるところ、このようなメッシュを、特許文献１の培養容器ように、ポリエチレン系樹脂を材料とする可撓性素材を用いて形成した容器に取り付けるには、シール強度が弱いという問題があった。
　そこで、かかる問題点に着眼して、本出願人が、さらなる鋭意検討を重ねた結果、本発明を完成するに至った。 However, in order to use as a filter member, a mesh made of polyester or polyamide which is easily available with a mesh size small enough not to allow passage of cells is suitable. In order to attach to a container formed using a flexible material made of polyethylene resin, such as a culture container, there is a problem that the sealing strength is weak.
Accordingly, as a result of further intensive studies by the present applicant, focusing on such problems, the present invention has been completed.
　すなわち、本発明は、細胞培養に際して行われる細胞や培地の分画、ろ過、洗浄などの種々の操作を簡便、かつ、効率良く行うために、容器本体の内部をフィルタ部材によって区画するにあたり、フィルタ部材とのシール強度が高められたフィルタ付き細胞用バッグ、及びその製造方法の提供を目的とする。 That is, 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.
　本発明に係るフィルタ付き細胞用バッグは、容器本体と注入出用ポートとを備え、前記容器本体の内部が、フィルタ部材によって区画されて、少なくとも一つの区画に注入出用ポートが設けられており、前記フィルタ部材が、目開き２０～３００μｍのポリエステル製又はポリアミド製のメインメッシュと、その表裏両面に配された目開き１００～４０００μｍのポリエチレン製のサブメッシュとからなり、前記容器本体が、少なくともポリエチレン層を含む可撓性フィルムの間に、前記ポリエチレン層を対向させて、前記フィルタ部材を挟んで周縁部を熱融着してなり、前記容器本体の周縁部に形成される熱融着部において、前記ポリエチレン製のサブメッシュが溶融して、前記メインメッシュの空隙部に侵入しつつ互いに融着されているともに、前記可撓性フィルムの前記ポリエチレン層と融着している構成としてある。 The cell bag with a filter according to the present invention 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 μm, and a polyethylene sub-mesh having a mesh opening of 100 to 4000 μm 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.
　また、本発明に係るフィルタ付き細胞用バッグの製造方法は、容器本体と注入出用ポートとを備え、前記容器本体の内部が、フィルタ部材によって区画されて、少なくとも一つの区画に注入出用ポートが設けられたフィルタ付き細胞用バッグの製造方法であって、目開き２０～３００μｍのポリエステル製又はポリアミド製のメインメッシュの表裏両面に、目開き１００～４０００μｍのポリエチレン製のサブメッシュを配して前記フィルタ部材を用意し、少なくともポリエチレン層を含む可撓性フィルムの間に、前記ポリエチレン層を対向させて、前記フィルタ部材を挟み、これらの周縁部を熱融着する方法としてある。 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 μm is provided on both sides of a main mesh made of polyester or polyamide having an opening of 20 to 300 μm. 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.
　本発明によれば、容器本体を形成する可撓性フィルムとフィルタ部材とのシール強度が高められ、破袋強度が向上したフィルタ付き細胞用バッグを提供することができる。 According to 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.
本発明の実施形態に係るフィルタ付き細胞用バッグの一例を示す説明図である。It is explanatory drawing which shows an example of the cell bag with a filter which concerns on embodiment of this invention. 本発明の実施形態に係るフィルタ付き細胞用バッグの製造例を模式的に示す説明図である。It is explanatory drawing which shows typically the manufacture example of the cell bag with a filter which concerns on embodiment of this invention. 熱融着部における可撓性フィルム、メインメッシュ、サブメッシュのそれぞれの端縁の位置関係を示す説明図である。It is explanatory drawing which shows the positional relationship of each edge of a flexible film, a main mesh, and a submesh in a heat-fusion part. 本発明の実施形態に係るフィルタ付き細胞用バッグの一使用例を示す説明図である。It is explanatory drawing which shows one usage example of the cell bag with a filter which concerns on embodiment of this invention.
　以下、本発明の好ましい実施形態について、図面を参照しつつ説明する。
　なお、図１は、本実施形態に係るフィルタ付き細胞用バッグの一例を示す説明図であり、図１(ａ)は、フィルタ付き細胞用バッグ１の平面図、図１（ｂ）は、フィルタ付き細胞用バッグ１の側面図、図１（ｃ）は、図１（ａ）のＡ－Ａ断面図である。 Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.
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, and FIG. 1 (b) is a filter. A side view of the attached cell bag 1, FIG. 1C is a cross-sectional view taken along the line AA of FIG.
　本実施形態の一例として図１に示すフィルタ付き細胞用バッグ１は、容器本体２と注入出用ポート３ａ，３ｂとを備えている。そして、容器本体２の内部は、フィルタ部材４によって二室に区画されており、それぞれの区画（第一区画１ｓｔ及び第二区画２ｎｄ）に注入出用ポート３ａ，３ｂが設けられている。 As an example of this embodiment, 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).
　注入出用ポート３ａ，３ｂは、培地や細胞を注入、排出などする際の出入り口となる部位であり、例えば、培地や細胞などが流通可能な管状の部材を容器本体２に取り付けることによって設けることができる。注入出用ポート３ａ，３ｂを形成する管状の部材は、例えば、ポリエチレン、ポリプロピレン、塩化ビニル、ポリスチレン系エラストマー、ＦＥＰなどの熱可塑性樹脂を用いて、射出成形、押出成形などにより、所定の形状に成形することができる。 The injection ports 3a and 3b are portions that serve as entrances and exits when injecting and discharging culture media and cells. For example, 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. Can do. 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.
　フィルタ部材４は、メッシュサイズが異なる二種類のメッシュを用いて構成され、メッシュサイズが相対的に小さいメインメッシュ４ａと、その表裏両面に配されたメッシュサイズが相対的に大きいサブメッシュ４ｂとからなっている。
　メインメッシュ４ａには、ポリエチレンテレフタレート等のポリエステル系樹脂繊維を織製したポリエステル製メッシュ、又はナイロン等のポリアミド系樹脂繊維を織製してなるポリアミド製メッシュが用いられる。サブメッシュ４ｂには、ポリエチレン系樹脂繊維を織製してなるポリエチレン製メッシュが用いられる。
　なお、サブメッシュ４ｂを形成するポリエチレン系樹脂繊維は、後述する可撓性フィルム２０のポリエチレン層２０ａと同種のポリエチレン系樹脂からなるのが好ましい。 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.
As the main mesh 4a, 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.
In addition, it is preferable that 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.
　これらのメッシュ４ａ，４ｂのメッシュサイズは、フィルタ付き細胞用バッグ１の使用態様に応じて適宜選定することができるが、メインメッシュ４ａについては目開き２０～３００μｍの範囲で、サブメッシュ４ｂについては目開き１００～４０００μｍの範囲で、これらのメッシュサイズを選定する。このとき、サブメッシュ４ｂの目開きが、メインメッシュ４ａの目開きに対して２～１０倍の大きさとなるように、上記範囲から選定するのが好ましい。
　なお、フィルタ付き細胞用バッグ１の使用例については後述する。 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 μm, and the submesh 4b has a mesh size of 20 to 300 μm. These mesh sizes are selected in the range of 100 to 4000 μm. 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.
In addition, the usage example of the cell bag 1 with a filter is mentioned later.
　容器本体２は、可撓性フィルム２０をバッグ状に製袋することによって形成される。
　容器本体２を形成する可撓性フィルム２０としては、少なくともポリエチレン層２０ａを含む可撓性フィルム２０が用いられる。かかる可撓性フィルム２０は、ポリエチレン層２０ａを単層で含むポリエチレン系樹脂フィルムの他、例えば、ポリプロピレン、エチレン－酢酸ビニル共重合体、ポリエステル、ポリアミド、シリコーン系エラストマー、ポリスチレン系エラストマー、テトラフルオロエチレン－ヘキサフルオロプロピレン共重合体（ＦＥＰ）などの熱可塑性樹脂からなる一種又は二種以上の層に、ポリエチレン層２０ａが表層に位置するように積層された積層フィルムであってもよい。
　なお、容器本体２を形成する可撓性フィルム２０は、細胞培養の進行状況や内部の細胞の状態又は培地の色などを確認できる程度の透明性を有しているのが好ましい。 The container body 2 is formed by making the flexible film 20 into a bag shape.
As the flexible film 20 forming the container body 2, a flexible film 20 including at least a polyethylene layer 20 a is used. Such 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. -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.
In addition, it is preferable that 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.
　容器本体２は、例えば、所望のサイズに裁断された二枚の可撓性フィルム２０を揃えて重ね合せ、その際、二枚の可撓性フィルム２０の間に、そのポリエチレン層２０ａを対向させて、フィルタ部材４を挟み込んだ状態で、これらの周縁部を熱融着することによって形成することができる。
　このとき、フィルタ部材４によって区画された、それぞれの区画１ｓｔ，２ｎｄに注入出用ポート３ａ，３ｂを設けるには、容器本体２の周縁部を熱融着する際に、容器本体２の周縁部における可撓性フィルム２０とフィルタ部材４との間の所定の位置に、注入出用ポート３ａ，３ｂを形成する管状の部材を挟んで熱融着するようにすればよい。
　また、熱融着に際しては、その温度は、サブメッシュ４ｂに用いたポリエチレン系樹脂の融点以上、メインメッシュ４ａに用いたポリエステル系樹脂又はポリアミド系樹脂の融点未満の温度とする。
　なお、図２は、フィルタ付き細胞用バッグ１の製造例を模式的に示す説明図であり、可撓性フィルム２０、フィルタ部材４、注入出用ポート３ａ，３ｂの配置を示している。 For example, 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.
At this time, in order to provide the injection ports 3a and 3b in the sections 1st and 2nd partitioned by the filter member 4, when the peripheral part of the container body 2 is heat-sealed, the peripheral part of the container body 2 What is necessary is just to heat-seal | push the tubular member which forms the ports 3a and 3b for injection | pouring in the predetermined position between the flexible film 20 and the filter member 4 in this.
In heat sealing, the temperature is higher than the melting point of the polyethylene resin used for the sub-mesh 4b and lower than the melting point of the polyester resin or polyamide resin used for the main mesh 4a.
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.
　このようにして形成された容器本体２にあっては、その周縁部に形成される熱融着部５において、メインメッシュ４ａの表裏両面に配されたサブメッシュ４ｂ（具体的には、サブメッシュ４ｂを形成するポリエチレン系樹脂繊維）が溶融して、メインメッシュ４ａの空隙部に侵入しつつ互いに融着されているとともに、可撓性フィルム２０のポリエチレン層２０ａと融着している。その結果、容器本体２を形成する可撓性フィルム２０とフィルタ部材４とのシール強度が高まり、フィルタ付き細胞用バッグ１の破袋強度が向上する。 In the container body 2 formed as described above, 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.
　可撓性フィルム２０の間にフィルタ部材４を挟み込んだ状態で、これらの周縁部を熱融着して容器本体２を製袋するにあたり、フィルタ部材４についても所望のサイズに裁断するが、熱融着部５において、メインメッシュ４ａの端縁が、サブメッシュ４ｂの端縁よりも容器内方に位置するように切り揃えておくことが好ましい。さらに、サブメッシュ４ｂの端縁が、容器本体２を形成する可撓性フィルム２０の端縁よりも容器内方に位置するように切り揃えておくのがより好ましい。
　なお、図３は、図１（ａ）のＢ－Ｂ断面に相当し、熱融着部５における可撓性フィルム２０、メインメッシュ４ａ、サブメッシュ４ｂのそれぞれの端縁の位置関係を示し、可撓性フィルム２０の端縁を一点鎖線αで示し、メインメッシュ４ａの端縁を一点鎖線βで示し、サブメッシュ４ｂの端縁を一点鎖線γで示している。 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. In the fusion | 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. 1 (a), and shows the positional relationship of the respective edges of the flexible film 20, the main mesh 4a, and the submesh 4b in the heat-sealed portion 5. An end edge of the flexible film 20 is indicated by a one-dot chain line α, an end edge of the main mesh 4a is indicated by a one-dot chain line β, and an end edge of the sub-mesh 4b is indicated by a one-dot chain line γ.
　このようにすることで、熱融着部５の最外縁部では、可撓性フィルム２０のポリエチレン層２０ａどうしが直接、又はサブメッシュ４ｂを介して、これらが溶融して融着するようになる。その結果、容器本体２の最外縁部がより強固に熱融着されて、フィルタ付き細胞用バッグ１の破袋強度がより一層向上する。 By doing in this way, in the outermost edge part of the heat-fusion part 5, the polyethylene layers 20a of the flexible film 20 are melted and fused directly or via the sub-mesh 4b. . As a result, 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.
　次に、本実施形態の一例として図１に示すフィルタ付き細胞用バッグ１の使用例について説明する。
　図４は、図１に示すフィルタ付き細胞用バッグ１を、培地交換などの操作を簡便、かつ、効率良く行うことができる培養バッグとして使用する例を示している。 Next, the usage example of the cell bag 1 with a filter shown in FIG. 1 is demonstrated as an example of this embodiment.
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.
　本使用例では、培地で満たされた容器本体２内に、第二区画２ｎｄに設けた注入出用ポート３ｂから、培養対象の細胞Ｃを注入して培養を行う。このとき、培養対象の細胞Ｃの大きさを考慮して、フィルタ部材４のメインメッシュ４ａとして、かかる細胞Ｃの通過を許容しないメッシュサイズのものを選択する。これにより、培養中の細胞Ｃは、第二区画２ｎｄに留まり、第一区画１ｓｔには移動しないようにすることができる（図４（ａ）参照）。
　なお、フィルタ部材４のサブメッシュ４ｂのメッシュサイズは、メインメッシュ４ａのメッシュサイズよりも大きければよく、培養対象の細胞Ｃの大きさによらずに適宜選択できる。 In this usage example, 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. At this time, considering the size of the cell C to be cultured, 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. Thereby, the cell C in culture can remain in the second section 2nd and not move to the first section 1st (see FIG. 4A).
In addition, 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 | size of the cell C to be cultured.
　培養期間中に培地交換を行う際には、第一区画１ｓｔに設けた注入出用ポート３ａから、古い培地を排出する（図４（ｂ）参照）。このとき、培養中の細胞Ｃを第二区画２ｎｄに留まらせることができるため、古い培地と一緒に排出されないようにすることができ、古い培地をより多く排出させることが可能になる。古い培地を排出した後は、それと同量の新しい培地を第一区画１ｓｔに設けた注入出用ポート３ａから注入し（図４（ｃ）参照）、培養を継続する。 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.
　このように、本使用例にあっては、培養期間中の培地交換、特に、古い培地の排出操作を簡便、かつ、効率良く行うことができる。 Thus, in this use example, the medium exchange during the culture period, in particular, the operation of discharging the old medium can be performed easily and efficiently.
　また、本使用例におけるフィルタ付き細胞用バッグ１は、第二区画２ｎｄに細胞Ｃを留まらせた状態で、第一区画１ｓｔに設けた注入出用ポート３ａから、容器内の内容液を排出することができるため、例えば、特許文献１の細胞培養用キットの培養容器として使用すれば、特許文献１が例示する細胞の回収操作も、細胞の沈降に時間をかけずに、簡便、かつ、効率良く行うことができる。さらに、特許文献１の細胞培養用キットの細胞回収容器として使用し、培養容器から回収された細胞懸濁液を第二区画２ｎｄに移送して、第一区画１ｓｔに設けた注入出用ポート３ａから、洗浄液の注入、排出を繰り替えして細胞の洗浄操作が行われるようにすることで、特許文献１の細胞培養用キットに洗浄回収機構を導入することができる。 In addition, 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. Furthermore, it is used as a cell collection container of the cell culture kit of Patent Document 1, and the cell suspension recovered from the culture container is transferred to the second compartment 2nd, and the injection port 3a provided in the first compartment 1st Therefore, 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.
　このように、本使用例では、フィルタ部材４のメインメッシュ４ａとして、培養対象の細胞Ｃの通過を許容しないメッシュサイズのものを選択して、フィルタ部材４によって区画された一方の区画２ｎｄに細胞Ｃを注入して培養を行うことで、当該区画２ｎｄに細胞Ｃを留まらせた状態で、フィルタ部材４によって区画された他方の区画１ｓｔに設けた注入出用ポート３ａから、培地や洗浄液などを注入出することができる。 Thus, in this usage example, as the main mesh 4a of the filter member 4, 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. By injecting C and culturing, in a state where the cells C remain in the section 2nd, 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.
　本使用例は、本実施形態に係るフィルタ付き細胞用バッグ１の一使用例であり、本実施形態に係るフィルタ付き細胞用バッグ１を細胞培養の用途に供することで、細胞培養に際して行われる細胞の分画、ろ過、洗浄、又は培地のろ過などの種々の操作を簡便、かつ、効率良く行うことができる。 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.
　以上、本発明について、好ましい実施形態を示して説明したが、本発明は、前述した実施形態に限定されるものではなく、本発明の範囲で種々の変更実施が可能であることはいうまでもない。 Although the present invention has been described with reference to the preferred embodiments, the present invention is not limited to the above-described embodiments, and various modifications can be made without departing from the scope of the present invention. Absent.
　例えば、前述した実施形態では、容器本体２の内部を二室に区画した例を示したが、同様のフィルタ部材４を複数用いて、三室以上に区画することもできる。また、それぞれの区画に、注入出用ポート３ａ，３ｂを一つずつ設けた例を示したが、必要に応じて、二以上の注出入用ポートを設けてもよい。また、注出入用ポートは、少なくとも一つの区画に設けられていればよく、注出入用ポートが設けられていない区画があってもよい。 For example, in the above-described embodiment, an example in which the inside of the container body 2 is partitioned into two chambers has been shown, but a plurality of similar filter members 4 may be used to partition into three or more chambers. Moreover, although the example which provided each one port 3a, 3b for injection | pouring in each division was shown, you may provide two or more pouring / injecting ports as needed. In addition, 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.
　この明細書に記載の文献及び本願のパリ優先権の基礎となる日本出願明細書の内容を全てここに援用する。 All the contents of the documents described in this specification and the content of the Japanese application that forms the basis of the Paris priority of the present application are incorporated herein.
　本発明は、医薬学・生化学分野において、細胞を効率良く培養する技術として利用できる。 The present invention can be used as a technique for efficiently culturing cells in the fields of pharmaceutical science and biochemistry.
　１　　　　　フィルタ付き細胞用バッグ
　２　　　　　容器本体
　２０　　　　　可撓性フィルム
　２０ａ　　　　　ポリエチレン層
　３ａ，３ｂ　　　　　注入出用ポート
　４　　　　　フィルタ部材
　４ａ　　　　　メインメッシュ
　４ｂ　　　　　サブメッシュ
　５　　　　　熱融着部
　１ｓｔ　　　　　第一区画
　２ｎｄ　　　　　第二区画 DESCRIPTION OF SYMBOLS 1 Cell bag with a filter 2 Container main body 20 Flexible film 20a Polyethylene layer 3a, 3b Port for injection | emission 4 Filter member 4a Main mesh 4b Submesh 5 Thermal fusion part 1st 1st division 2nd 2nd division

Claims (3)

  1. 　容器本体と注入出用ポートとを備え、
    　前記容器本体の内部が、フィルタ部材によって区画されて、少なくとも一つの区画に注入出用ポートが設けられており、
    　前記フィルタ部材が、目開き２０～３００μｍのポリエステル製又はポリアミド製のメインメッシュと、その表裏両面に配された目開き１００～４０００μｍのポリエチレン製のサブメッシュとからなり、
    　前記容器本体が、少なくともポリエチレン層を含む可撓性フィルムの間に、前記ポリエチレン層を対向させて、前記フィルタ部材を挟んで周縁部を熱融着してなり、
    　前記容器本体の周縁部に形成される熱融着部において、前記ポリエチレン製のサブメッシュが溶融して、前記メインメッシュの空隙部に侵入しつつ互いに融着されているともに、前記可撓性フィルムの前記ポリエチレン層と融着していることを特徴とするフィルタ付き細胞用バッグ。     It has a container body and an injection port,
    The inside of the container body is partitioned by a filter member, and an injection port is provided in at least one partition,
    The filter member comprises a main mesh made of polyester or polyamide having a mesh opening of 20 to 300 μm, and a polyethylene sub-mesh having a mesh opening of 100 to 4000 μm arranged on both the front and back sides.
    The container body is formed by heat-sealing a peripheral portion with the filter member sandwiched between the flexible film including at least a polyethylene layer,
    In the heat-sealed portion formed at the peripheral portion of the container main body, the polyethylene sub-mesh is melted and fused with each other while invading into the gap of the main mesh, and the flexible film A cell bag with a filter, which is fused to the polyethylene layer.
  2. 　前記熱融着部において、前記メインメッシュの端縁が、前記サブメッシュの端縁よりも容器内方に位置し、前記熱融着部の最外縁部では、前記可撓性フィルムの前記ポリエチレン層どうしが直接、又は前記サブメッシュを介して、これらが溶融して融着している請求項１に記載のフィルタ付き細胞用バッグ。 In the heat fusion part, the edge of the main mesh is located inside the container from the edge of the sub-mesh, and the outermost edge part of the heat fusion part is the polyethylene layer of the flexible film. The cell bag with a filter according to claim 1, wherein these are melted and fused directly or via the sub-mesh.
  3. 　容器本体と注入出用ポートとを備え、前記容器本体の内部が、フィルタ部材によって区画されて、少なくとも一つの区画に注入出用ポートが設けられたフィルタ付き細胞用バッグの製造方法であって、
    　目開き２０～３００μｍのポリエステル製又はポリアミド製のメインメッシュの表裏両面に、目開き１００～４０００μｍのポリエチレン製のサブメッシュを配して前記フィルタ部材を用意し、
    　少なくともポリエチレン層を含む可撓性フィルムの間に、前記ポリエチレン層を対向させて、前記フィルタ部材を挟み、
    　これらの周縁部を熱融着することを特徴とするフィルタ付き細胞用バッグの製造方法。     A method for producing a cell bag with a filter comprising a container body and an injection port, wherein the inside of the container body is partitioned by a filter member, and an injection port is provided in at least one partition,
    The filter member is prepared by arranging a polyethylene sub-mesh having a mesh size of 100 to 4000 μm on both sides of a main mesh made of polyester or polyamide having a mesh size of 20 to 300 μm,
    Between the flexible film including at least a polyethylene layer, the polyethylene layer is opposed to the filter member,
    A method for producing a cell bag with a filter, characterized in that these peripheral portions are heat-sealed.
PCT/JP2017/044925 2016-12-26 2017-12-14 Filter-equipped bag for cells and production method therefor WO2018123619A1 (en)

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