Cell container, method for producing same, port member, and method for culturing cells

Abstract

This cell container comprises: a resin bag member; and a port member which includes a molded body of a resin composition and communicates the inside and the outside of the bag member. The bag member includes a multilayer resin film which includes a surface layer, a core layer, and a heat seal layer in this order from the outside toward the inside of the cell container, and the heat seal layer contains a 4-methyl-1-pentene-based copolymer.

Description

Cell container and method for manufacturing the same, port member, and method for culturing cells

[0001] The present invention relates to a cell container and a method for manufacturing the same, a port member, and a method for culturing cells.

[0002] Known cell containers for growing, growing, storing, and transporting cells include a cell container in which an oxygen-permeable film is heat-sealed to form a sealed bag member, and a port member attached to a part of the bag connects the inside and outside of the container (see, for example, Patent Document 1). Patent Document 1 describes that the port member is sandwiched and welded between the peripheral edges of two gas-permeable films, both of which are planar articles.

[0003] International Publication No. 2022 / 014436

[0004] As described in Patent Document 1, cell containers are sometimes provided with a port member for introducing and retrieving culture fluid and cultured cells. However, when welding to a bag member, depending on the combination of material of the members, welding between the bag member and the port member may be insufficient, making it impossible to ensure airtightness of the cell container.

[0005] The present invention has been made in consideration of the above-mentioned problems of the conventional technology, and aims to provide a cell container having a bag member and a port member that can improve adhesion, a method for manufacturing the same, a port member used in the cell container, and a method for culturing cells using the cell container.

[0006] One aspect of the present invention for solving the above problems relates to the following cell containers [1] to

[16] . [1] A cell container having a resin bag member and a port member that communicates between the inside and outside of the bag member and includes a molded article of a resin composition, wherein the bag member includes a multilayer resin film that includes, from the outside to the inside of the cell container, a surface layer, a core layer, and a heat seal layer in this order, and the heat seal layer includes a 4-methyl-1-pentene copolymer. [2] The cell container according to [1], wherein the core layer includes a 4-methyl-1-pentene copolymer or a styrene-based elastomer. [3] The cell container according to [1] or [2], wherein the 4-methyl-1-pentene copolymer contained in the heat seal layer has 80 mol% to 99 mol% of structural units derived from 4-methyl-1-pentene, and 1 mol% to 20 mol% of structural units derived from an α-olefin having 2 to 20 carbon atoms, excluding structural units derived from 4-methyl-1-pentene, relative to all structural units. [4] The bag member has an oxygen permeability of 3.0 L / (m ２ ・day・atm) or more 100.0L / (m ２ [5] The cell container according to any one of [1] to [3], wherein the bag member is formed from the resin film having a sterility assurance level (SAL) of 10 or less as measured in accordance with BS EN556-1:2001. －３The cell container according to any one of [1] to [4], wherein the cell container is one of the following: [6] The cell container according to any one of [1] to [5], wherein the resin composition molded article contained in the port member is a resin composition molded article containing at least one resin selected from the group consisting of a 4-methyl-1-pentene copolymer, polyethylene, and polypropylene. [7] The port member according to any one of [1] to [6], wherein the resin composition molded article contained in the port member is a resin composition molded article having a peel strength of 20 N / 15 mm or more and 100 N / 15 mm or less as measured by a 180° peel test performed in accordance with the method described below. Measurement conditions: A 65 mm x 65 mm x 1.0 mm press sheet obtained from the resin composition is heat-sealed to the planar surface of a 65 mm x 65 mm x 0.3 mm substrate formed from a 4-methyl-1-pentene copolymer consisting of 4-methyl-1-pentene structural units and propylene structural units at a temperature of 240°C, a pressure of 0.1 MPa, a pressure time of 5 seconds, and a seal width of 10 mm, and then a 180° peel test is performed using a tensile tester at 23°C. [8] The cell container according to any of [1] to [7], wherein the molded article of the resin composition contained in the port member is a molded article of a resin composition having a total light transmittance of 85% or more when measured on a 1 mm thick press sheet. [9] The cell container according to any one of [1] to [8], wherein the molded article of the resin composition contained in the port member is a molded article of the resin composition having a durometer hardness type D measured in accordance with ASTM D2240 of 30 or more and 80 or less when measured on a 1 mm thick press sheet.

[10] The cell container according to any one of [1] to [9], wherein the port member has a narrow tubular portion having a shorter circumferential length and a thick tubular portion having a longer circumferential length, and at least the thick tubular portion contains the molded article of the resin composition.

[11] The cell container according to any one of [1] to

[10] , wherein the port member is joined to the bag member via a single-layer or multi-layer thermal adhesive film having at least an elastomer layer containing a styrene-based elastomer.

[12] The cell container according to

[11] , wherein the thermal adhesive film is disposed so as to surround the circumferential direction of the cylindrical port member, and the elastomer layer is disposed so as to surround the entire periphery of the port member.

[13] The cell container according to

[12] , wherein the thermal adhesive film is welded to the entire periphery of the port member.

[14] The cell container according to any one of

[11] to

[13] , wherein the thermal adhesive film has a layer containing 4-methyl-1-pentene copolymer.

[15] The cell container according to

[14] , wherein the layer containing 4-methyl-1-pentene copolymer is a sealant layer.

[16] The cell container according to any one of

[11] to

[15] , wherein the bonding width of the thermal adhesive film to the port member in the direction in which the port member communicates between the inside and outside of the bag member is 1 mm or more and 50 mm or less.

[0007] One aspect of the present invention for solving the above problems relates to the following methods for manufacturing a cell container

[17] and

[18] .

[17] A method for manufacturing a cell container, comprising the steps of: arranging bag members made of resin containing at least a 4-methyl-1-pentene copolymer with their ends overlapping; and heat-sealing the overlapped bag members to form a bag, wherein in the overlapping and arranging step, a port member is placed between the overlapped ends; and in the processing step, the bag members with the port member placed therebetween are heated and pressurized to bond the bag members to the port member.

[18] The method for manufacturing a cell container according to

[17] , wherein in the overlapping and arranging step, a single-layer or multi-layer thermal adhesive film having an elastomer layer containing at least a styrene-based elastomer is placed between the port member and the bag members, and in the processing step, the bag member and the port member are bonded via the thermal adhesive film.

[0008] One aspect of the present invention for solving the above problems relates to a port member as set forth in

[19] below:

[19] A port member comprising a resin bag member containing at least a 4-methyl-1-pentene copolymer, and a port member welded to the resin bag member to communicate between the inside and outside of the bag member, the port member comprising a molded article of a resin composition containing the 4-methyl-1-pentene copolymer, wherein the resin composition has a total light transmittance of 85% or more when measured on a pressed sheet having a thickness of 1 mm.

[0009] One aspect of the present invention for solving the above problems relates to a method for culturing cells according to the following item

[20] :

[20] The method for culturing cells comprises the steps of: introducing cells into a cell container according to any one of items [1] to

[16] ; and culturing the cells in the cell container.

[0010] According to the present invention, there are provided a cell container having a bag member and a port member that can improve adhesion, a method for manufacturing the same, a port member used in the cell container, and a method for culturing cells using the cell container.

[0011] FIG. 1A is a plan view showing the appearance of a cell container having a port member, and FIG. 1B is a partial cross-sectional view of the cell container shown in FIG. 1A taken along dashed line 1B-1B. FIG. 2A is a plan view showing the appearance of a cell container according to one embodiment of the present invention, and FIG. 2B is a partial cross-sectional view of the cell container shown in FIG. 2A taken along dashed line 2B-2B. FIG. 3A is a plan view of a port member 200, FIG. 3B is a front view of the port member as viewed from the inside to the outside of the bag member, and FIG. 3C is a rear view of the port member as viewed from the outside to the inside of the bag member. FIG. 4A is an exemplary cross-sectional view of a single-layer thermal adhesive film, FIG. 4B is an exemplary cross-sectional view of a two-layer thermal adhesive film, and FIG. 4C is an exemplary cross-sectional view of a three-layer thermal adhesive film. FIGS. 5A to 5C are schematic diagrams showing the width of the thermal adhesive film in the width direction of the joint.

[0012] 1. Cell Container Figure 1A is a plan view showing the appearance of a cell container according to one embodiment of the present invention, and Figure 1B is a partial cross-sectional view of the cell container shown in Figure 1A taken along dashed line 1B-1B. Figure 2A is a plan view showing the appearance of a cell container according to one embodiment of the present invention, and Figure 2B is a partial cross-sectional view of the cell container shown in Figure 2A taken along dashed line 2B-2B.

[0013] The cell container 100 is a container used to culture cells by introducing a culture medium and cells into the container. In this specification, culturing cells means multiplying, growing, or maintaining the cells in a living state.

[0014] The cell container 100 is formed by overlapping two resin films 112 and 114 and heat-sealing the resin films around the entire periphery of the edges to form a joint 116, thereby forming a substantially sealed bag member 110. One or more port members 200 (three in this embodiment) are sandwiched between the resin films 112 and 114 in a portion of the heat-sealed joint 116. The port members 200 may be joined in direct contact with the resin films 112 and 114 as shown in FIGS. 1A and 1B, or may be joined to these resin films by heat sealing at least partially via a thermal adhesive film 130 as shown in FIGS. 2A and 2B.

[0015] 1-1. Bag Member 110 The bag member 110 is formed by welding together a resin film 112 and a resin film 114. Both the resin film 112 and the resin film 114 are multilayer thermoplastic resin films formed by molding a thermoplastic resin composition containing a 4-methyl-1-pentene copolymer into a film shape. From the viewpoint of improving the heat seal strength and the gas permeability to oxygen and carbon dioxide, at least one of the resin film 112 and the resin film 114 contains a 4-methyl-1-pentene copolymer. It is preferable that at least one of the resin film 112 and the resin film 114 further contains a styrene-based elastomer or a butene-based copolymer.

[0016] In this embodiment, both the resin film 112 and the resin film 114 are laminates of at least three layers, each having a surface layer and a heat seal layer disposed on the outermost surfaces of both sides of the film, and a core layer sandwiched therebetween. These layers are disposed in the order of the surface layer, the core layer, and the heat seal layer from the outside to the inside of the cell container 100.

[0017] The surface layer and the heat seal layer can be used as support layers for sandwiching and holding the core layer from above and below. The heat seal layer is used for heat sealing with other resin films. The surface layer and the heat seal layer can also be used as layers for suppressing blocking when a soft, highly oxygen-permeable core layer is overlaid, thereby enhancing the anti-blocking properties of the resin film.

[0018] The thickness of the surface layer and the heat seal layer is preferably 5 μm or more and 300 μm or less, more preferably 5 μm or more and 200 μm or less, and even more preferably 5 μm or more and 150 μm or less. The thicker the surface layer and the heat seal layer, the more the heat sealability of the resin film can be improved. Furthermore, the thinner the thickness of the surface layer and the heat seal layer, the more the oxygen permeability of the bag member can be improved.

[0019] The thickness of the core layer is preferably 10 μm or more and 0.5 mm or less, more preferably 15 μm or more and 0.3 mm or less, and even more preferably 20 μm or more and 100 μm or less. The thicker the core layer, the more the flatness of the film can be improved. By appropriately thinning the thickness of the core layer, the oxygen permeability of the bag member can be improved.

[0020] The resin film contains a 4-methyl-1-pentene copolymer in at least one of these layers. This can increase the oxygen permeability of the resin film. The resin film may contain a 4-methyl-1-pentene copolymer in multiple layers. In this case, the 4-methyl-1-pentene copolymer contained in each layer may be a copolymer having the same physical properties such as copolymerization ratio, or may be a different copolymer having different physical properties such as copolymerization ratio.

[0021] The resin film preferably contains at least one thermoplastic resin or thermoplastic elastomer selected from the group consisting of ethylene-based (co)polymers, propylene-based (co)polymers, butene-based (co)polymers, and styrene-based elastomers in at least one of these layers. By using an appropriate combination of these thermoplastic resins or thermoplastic elastomers, the transparency, oxygen permeability, and heat seal strength of the resin film can be well balanced.

[0022] In this embodiment, the resin film contains a 4-methyl-1-pentene-propylene copolymer in at least the heat seal layer. This increases the heat seal strength of the resin film, as well as the gas permeability to oxygen and carbon dioxide, and also increases the anti-blocking properties between films. Furthermore, the resin film preferably contains a butene copolymer in at least the heat seal layer. This increases the heat sealability and transparency of the resin film.

[0023] The resin film preferably contains a thermoplastic elastomer such as a polyolefin elastomer or a styrene elastomer in the core layer. Hydrogenated SEBS, SEPS, and SEEPS can be preferably used as the styrene elastomer. Of these, polyolefin elastomers are preferred, and 4-methyl-1-pentene copolymers are more preferred, from the viewpoint of improving the flatness and oxygen permeability of the resin film. In this specification, the term "thermoplastic elastomer" refers to a polymer that exhibits fluidity when heated above its melting point if crystalline or above its glass transition point if amorphous, while exhibiting rubber elasticity at room temperature.

[0024] For example, the resin film may be a multilayer film having a surface layer and a heat seal layer each containing a 4-methyl-1-pentene copolymer and a butene copolymer, and a core layer containing a 4-methyl-1-pentene copolymer or a styrene elastomer.

[0025] The ethylene-based (co)polymer may be a homopolymer of ethylene or a copolymer of ethylene and an α-olefin having from 3 to 20 carbon atoms. Of these, a copolymer of ethylene and an α-olefin having from 3 to 20 carbon atoms is preferred. From the viewpoint of improving gas permeability and heat sealability, the α-olefin having from 3 to 20 carbon atoms is preferably propylene, butene, hexene, or octene. The proportion of structural units derived from ethylene to all structural units of the ethylene-based (co)polymer is preferably from 80 mol% to 95 mol%, more preferably from 85 mol% to 95 mol%, and even more preferably from 88 mol% to 93 mol%. The proportion of structural units derived from α-olefins having from 3 to 20 carbon atoms to all structural units of the ethylene-based (co)polymer is preferably from 5 mol% to 20 mol%, more preferably from 5 mol% to 15 mol%, and even more preferably from 7 mol% to 12 mol%.

[0026] The ethylene (co)polymer may also be a block polymer of ethylene and silicone.

[0027] The block copolymer may be a (polyethylene)-(silicone) diblock copolymer or a (polyethylene)-(silicone)-(polyethylene) terblock copolymer. The polyethylene may be an ethylene homopolymer or a copolymer of ethylene and an α-olefin having from 3 to 20 carbon atoms, but is preferably an ethylene homopolymer. When the polyethylene is a copolymer, the molar ratio of ethylene to the other α-olefin (ethylene:other α-olefin) is preferably from 81:19 to 99:1, and more preferably from 90:10 to 99:1.

[0028] The block copolymer can be obtained by reacting polyethylene having a vinyl group at one end with a silicon-containing compound in the presence of a catalyst, specifically by a method described in WO 2012 / 098865 or the like.

[0029] For example, the block copolymer may have the following structure:

[0030] A-CH ２ -CH ２ -Si(CH ３ ) ２ —O—(Si(CH ３ ) ２ -O) ｉ -Si(CH ３ ) ２ -CH ２ -CH ２ -A: Each of the two As independently represents a polyethylene, and i represents an integer of 1 or greater. The polyethylene represented by each As preferably has a number average molecular weight (Mn) of 100 or greater and 500,000 or less, more preferably 500 or greater and 50,000 or less, and even more preferably 700 or greater and 10,000 or less. i is preferably 1 or greater and 1,000 or less, more preferably 1 or greater and 300 or less, and even more preferably 1 or greater and 50 or less.

[0031] The propylene-based (co)polymer may be a homopolymer of propylene, or a copolymer of propylene and ethylene or an α-olefin other than propylene. Of these, a copolymer of propylene and an α-olefin other than propylene is preferred. The α-olefin other than propylene is preferably ethylene or an α-olefin having from 4 to 20 carbon atoms, and from the viewpoint of improving gas permeability and heat sealability, ethylene, butene, hexene, and octene are preferred. The proportion of structural units derived from propylene to all structural units of the propylene-based (co)polymer is preferably 70 mol% to 95 mol%. The proportion of structural units derived from ethylene or an α-olefin other than propylene to all structural units of the propylene-based (co)polymer is preferably 5 mol% to 30 mol%.

[0032] The 4-methyl-1-pentene copolymer of the heat seal layer is a copolymer of 4-methyl-1-pentene and ethylene or an α-olefin other than 4-methyl-1-pentene. The 4-methyl-1-pentene copolymer is preferably a copolymer of 4-methyl-1-pentene and ethylene or an α-olefin other than 4-methyl-1-pentene, and more preferably a copolymer of 4-methyl-1-pentene and propylene. The proportion of structural units derived from 4-methyl-1-pentene to all structural units of the 4-methyl-1-pentene copolymer is preferably 80 mol% or more and 99 mol% or less. The proportion of structural units derived from ethylene or an α-olefin other than 4-methyl-1-pentene to all structural units of the 4-methyl-1-pentene copolymer is preferably 1 mol% or more and 20 mol% or less.

[0033] The 4-methyl-1-pentene copolymer of the core layer is a copolymer of 4-methyl-1-pentene and ethylene or an α-olefin other than 4-methyl-1-pentene. The 4-methyl-1-pentene copolymer is preferably a copolymer of 4-methyl-1-pentene and ethylene or a linear α-olefin other than 4-methyl-1-pentene having from 2 to 20 carbon atoms, and examples of linear α-olefins having from 3 to 20 carbon atoms include propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, 1-decene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, and 1-eicosene. Of these, a copolymer of 4-methyl-1-pentene and 1-hexadecene or 1-octadecene is preferred. The proportion of structural units derived from 4-methyl-1-pentene to all structural units of the 4-methyl-1-pentene copolymer is preferably 80 mol% or more and 99 mol% or less. The proportion of structural units derived from ethylene or an α-olefin other than 4-methyl-1-pentene to all structural units of the 4-methyl-1-pentene copolymer is preferably 1 mol% or more and 20 mol% or less.

[0034] The butene copolymer is preferably a copolymer of butene and ethylene or an α-olefin other than butene, and is preferably a copolymer of 1-butene and ethylene, or a copolymer of 1-butene and propylene. The butene copolymer preferably contains structural units derived from 1-butene in a proportion of its total structural units of 80 mol% to 99 mol%, more preferably 83 mol% to 99 mol%, and even more preferably 85 mol% to 99 mol%. Furthermore, the butene copolymer preferably contains structural units derived from ethylene or an α-olefin other than butene in a proportion of its total structural units of 1 mol% to 20 mol%, more preferably 1 mol% to 17 mol%, and even more preferably 1 mol% to 15 mol%.

[0035] Ethylene-based (co)polymers, propylene-based (co)polymers, 4-methyl-1-pentene-based copolymers, and butene-based (co)polymers can be produced by polymerizing monomers in the presence of known catalysts such as Ziegler-Natta catalysts and metallocene catalysts using known polymerization methods such as a gas phase method, a bulk method, or a slurry method. The monomers may be derived from biomass or fossil fuels. Furthermore, both biomass-derived and fossil fuel-derived monomers may be used.

[0036] The styrene-based elastomer may be a block copolymer (SBC) having a polystyrene block that forms a hard segment and a conjugated diene monomer block such as isoprene or butadiene that forms a soft segment, such as a hydrogenated styrene-butadiene-styrene block copolymer (HSBR), a styrene-ethylene-propylene-styrene block copolymer (SEPS), a styrene-ethylene-butene-styrene block copolymer (SEBS), or a styrene-ethylene-ethylene-propylene-styrene block copolymer (SEEPS).

[0037] The resin film 112 and the resin film 114 may contain additives such as a heat stabilizer, an anti-slip agent, an anti-blocking agent, an anti-fogging agent, a lubricant, a pigment, a dye, a plasticizer, an antioxidant, a hydrochloric acid absorber, an antioxidant, a crystal nucleating agent, a filler (inorganic filler, organic filler), and a softener.

[0038] At least one of the resin films 112 and 114 has an oxygen permeability of 3.0 L / (m ２ ・day・atm) or more 100.0L / (m ２ ·day·atm) or less, and ２ ・day・atm) or more 50.0L / (m ２ ·day·atm) or less, and 5.0 L / (m ２ ・day・atm) or more 30.0L / (m ２ It is more preferable that the oxygen permeability of the resin film is equal to or less than 1 / 2 day / atm. By increasing the oxygen permeability of the resin film, the efficiency of cell culture in the cell container can be increased.

[0039] At least one of the resin films 112 and 114 has a carbon dioxide permeability of 10 L / (m ２ ・day・atm) or more 400L / (m ２ ·day·atm) or less, and ２ ・day・atm) or more 300L / (m ２ The higher the carbon dioxide permeability, the higher the cell culture efficiency of the cell container.

[0040] The oxygen permeability and carbon dioxide permeability were measured in accordance with JIS K 7126-1:2006 using a differential pressure gas permeability measuring device (manufactured by Toyo Seiki Seisakusho) at a test temperature of 23°C and a test relative humidity of 0% RH, with the measurement area of ​​the resin film being 5 cm. ２ When the oxygen permeability or carbon dioxide permeability is expected to be large, an aluminum mask is placed on the measurement sample in advance to reduce the actual permeation area to 5.0 cm ２ It is more preferable to set the following.

[0041] At least one of the resin films 112 and 114 has a moisture permeability of 10 g / (m ２ ・day) or more 100g / (m ２ ·day) or less, and ２ ・day) or more 80g / (m ２ ·day) or less, and 14 g / (m ２ ・day) or more 60g / (m ２ By setting the moisture permeability in this range, the moisture content is less likely to lose moisture.

[0042] The moisture permeability is calculated in accordance with the isobaric method (cup-type gravimetric method) described in JIS Z 0208:2021 under condition B (test temperature 40° C., test relative humidity 90% RH).

[0043] At least one of the resin films 112 and 114 has a sterility assurance level (SAL) of 10 or more for medical devices measured in accordance with BS EN556-1:2001. －３ Preferably, it is 10 －６ Here, SAL indicates the degree of sterility of the sterilization target after the sterilization process, and is expressed as the probability of microorganisms being expected to exist per unit number of the sterilization target after sterilization. SAL is 10 －ｎ The number of viable bacteria per sterilized item is expressed as 10 －ｎ n is preferably 3 or more, and more preferably 6 or more.

[0044] From the viewpoint of improving cell observation efficiency, at least one of resin film 112 and resin film 114 preferably has high visible light transmittance and low haze. Specifically, the total light transmittance measured in accordance with JIS K 7361-1:1997 is preferably 50% to 100%, more preferably 70% to 100%, and even more preferably 90% to 100%. Furthermore, the haze measured in accordance with JIS K7136:2000 is preferably 0% to 20%, and more preferably 0% to 10%.

[0045] It is preferable that resin film 112 and resin film 114 have high welding strength when heat-sealed. Specifically, the welding strength when resin film 112 and resin film 114 are heat-sealed is preferably 5 N / 15 mm width or more and 300 N / 15 mm width or less, more preferably 10 N / 15 mm width or more and 200 N / 15 mm width or less, and even more preferably 20 N / 15 mm width or more and 200 N / 15 mm width or less.

[0046] To measure the welding strength when the resin film 112 and the resin film 114 are heat-sealed, two rectangular heat-sealing test pieces measuring 150 mm wide x 50 mm long are cut out from the resin film so that the longitudinal direction coincides with the MD (machine direction) direction of the resin film. Next, the two heat-sealing test pieces are overlapped so that the heat-sealing layers of the resin films face each other, and then heat-sealed using a heat-sealing tester (Tester Sangyo Co., Ltd., thermal gradient heat-sealing tester, model number TP-701-G) under the following conditions: the upper and lower temperatures of the heat-sealing bar (heat-sealing temperature) are both 180°C, the seal width is 10 mm, the seal pressure is 0.1 MPa, and the seal time is 2 seconds. The heat-sealing is performed so that the welding line is perpendicular to the MD direction of the resin film. The two welded resin films were then removed from the tester, and 15 mm wide strip test pieces were cut out in directions perpendicular and parallel to the weld line as face-to-face heat seal test pieces of the resin films. Using a tensile tester (Orientec Co., Ltd., Tensilon Universal Testing Machine, Model RTG-1250), the strip test pieces were peeled at a test temperature of 23°C, a chuck distance of 50 mm, and a tensile speed of 300 mm / min, and the maximum peel strength was measured, which was taken as the heat seal strength (unit: N / 15 mm). The heat seal strength was measured for three test pieces, and the average value was calculated.

[0047] Resin films 112 and 114 preferably have high welding strength when heat-sealing port member 200. Specifically, when port member 200 is heat-sealed to resin film 112 or resin film 114, the welding strength is preferably 10 N / 15 mm width or more and 100 N / 15 mm width or less, more preferably 15 N / 15 mm width or more and 100 N / 15 mm width or less, and even more preferably 20 N / 15 mm width or more and 100 N / 15 mm width or less.

[0048] To measure the weld strength when the port member 200 is heat-sealed to the resin films 112 and 114, rectangular heat-seal test pieces measuring 65 mm wide and 65 mm long are cut from 10 mm wide and 50 mm long sheets cut from these films. Next, a press sheet (65 mm x 65 mm x 1.0 mm) obtained from the resin composition used to make the port member 200 is heat-sealed, followed by a 180° peel test using a tensile tester. The heat-sealing conditions are as follows: the heat-sealing temperature is at least 10°C higher (e.g., 220-240°C) than the melting point of the resin composition with the highest melting point among the resin compositions used to make the resin films 112 and 114 and the resin composition used to make the port member 200; the temperature of the bottom of the heat-seal bar is 23°C; the seal width is 10 mm; the seal pressure is 0.1 MPa; and the seal time is 2-5 seconds. The weld line is aligned in the machine direction of the multilayer film. The heat seal strength is measured for three test pieces and the average value is calculated.

[0049] The thickness of the resin films 112 and 114 is preferably 50 μm or more and 1 mm or less, more preferably 50 μm or more and 500 μm or less, and even more preferably 50 μm or more and 300 μm or less. The thicker the resin films 112 and 114, the stronger the strength of the bag member 110 and the welding strength when heat-sealed can be increased. The thinner the resin films 112 and 114, the higher the permeability to oxygen and carbon dioxide gases can be increased.

[0050] 1-2. Port Member The port member 200 is a hollow member that connects the inside and outside of the sealed bag member 110. One end of the hollow port member 200 protrudes inward from the bag member 110 beyond the heat-sealed joint 116, and the other end protrudes outward from the bag member 110 beyond the heat-sealed joint 116. Through the port member 200, a culture medium can be introduced into the bag member 110, or conversely, a culture medium can be discharged from the bag member 110. Alternatively, through the port member 200, cells to be cultured can be introduced into the bag member 110, or conversely, cultured cells can be removed from the bag member 110.

[0051] The number of port members 200 is not limited and may be one or more. When there are more than one, the number may be three as shown in Figures 1A and 2A, or may be two, four, or more. The positions at which the port members 200 are arranged are also not limited. There may be multiple port members 200 that communicate between the inside and outside of the bag member 110 facing in the same direction, or there may be multiple port members 200 that communicate between the inside and outside of the bag member 110 facing in different directions (e.g., opposite directions).

[0052] FIG. 3A is a plan view of the port member 200 (a plan view viewed in the same direction as FIGS. 1A and 2A ), FIG. 3B is a front view of the port member 200 viewed from the inside to the outside of the bag member 110, and FIG. 3C is a rear view of the port member 200 viewed from the outside to the inside of the bag member 110.

[0053] As shown in FIGS. 3A to 3C , the port member 200 includes a resin main body 210 and a passage 220 that penetrates the interior of the main body 210 from front to back and allows the flow of liquid and cells. The main body 210 further includes a thin tube portion 212 that is a long, thin cylinder with a shorter outer periphery and a thick, short columnar portion 214 with a longer outer periphery. The passage 220 penetrates the thin tube portion 212 and the thick tube portion 214, communicating them from front to back, while maintaining the same diameter. In this embodiment, the cross-sectional outer periphery of the thin tube portion 212 is circular, and the cross-sectional outer periphery of the thick tube portion 214 is boat-shaped. However, the cross-sectional outer periphery shapes are not limited to these. The cross-sectional outer periphery of the thin tube portion 212 may be elliptical, oval, or polygonal, such as a diamond, and the cross-sectional outer periphery of the thick tube portion 214 may be circular, elliptical, oval, or polygonal, such as a diamond.

[0054] The thin tube portion 212 is disposed protruding from the bag member 110 and is connected to a cap for sealing the cell container 100, an injection port for injecting a drug or the like, or a tube for connecting to a Luer port outside the bag member 110. Therefore, the shape of the thin tube portion 212 may be any shape that allows connection thereto. The thick tube portion 214 is disposed at the joint 116 of the bag member 110 and is welded to the two resin films 112 and 114. In this embodiment, the thick tube portion 214 has a diamond-shaped cross section, with a short diagonal in the stacking direction of the two films and a long diagonal in the planar direction of the two films. This allows the upper and lower edges to contact the respective films over a longer range, increasing the weld area between the thick tube portion 214 and the resin films 112 and 114 and further increasing the heat seal strength of the port member 200.

[0055] The passage 220 passes through the thin cylindrical portion 212 and the thick cylindrical portion 214. The inner diameter of the passage 220 may be constant throughout the thin cylindrical portion 212 and the thick cylindrical portion 214, or may vary midway.

[0056] In this embodiment, the port member 200 is an integrally molded body formed by injection molding a resin composition containing the material of the port member 200. However, the thin cylindrical portion 212 and the thick cylindrical portion 214 may be fabricated separately and then welded or bonded together.

[0057] From the viewpoint of improving the visibility of the cell container, the port member 200 preferably has high transparency to visible light.

[0058] By making the port member 200 a molded body with high hardness, even when a port member 200 having grooves or screws in the thin cylindrical portion 212 is manufactured, it is less likely to deform during welding, and when a soft tube is fitted, leakage of the internal fluid from the fitting portion can be prevented.

[0059] The material of the port member 200 may be a resin composition containing polyethylene or polypropylene from the viewpoints of resistance to material degradation due to gamma ray irradiation in gamma ray sterilization and improving bondability via a thermal adhesive film. From these viewpoints, the polyethylene is preferably linear low-density polyethylene (LLDPE) or high-density polyethylene (HDPE). Furthermore, from the viewpoint of improving bondability by heat sealing via a thermal adhesive film, the polyethylene preferably has a melting point (Tm) measured by differential scanning calorimetry (DSC) of 80°C or higher and 140°C or lower, more preferably 110°C or higher and 140°C or lower, and even more preferably 120°C or higher and 140°C or lower. The polypropylene is preferably homopolypropylene (hPP) or random polypropylene (rPP). From the viewpoint of improving the bondability by heat sealing via the thermal adhesive film, the polypropylene preferably has a melting point (Tm) measured by DSC of 120° C. or higher and 165° C. or lower, and more preferably 135° C. or higher and 165° C. or lower. Furthermore, the surface of the port member 200 that comes into close contact with the thermal adhesive film 130 is preferably smooth to make it easier to ensure close contact.

[0060] Alternatively, the material of the port member 200 can be a resin composition containing a 4-methyl-1-pentene copolymer, or a resin composition containing a 4-methyl-1-pentene copolymer and a butene copolymer, from the viewpoint of increasing the heat seal strength and breaking elongation of the port member 200 to the resin films 112 and 114.

[0061] The 4-methyl-1-pentene copolymer is preferably a copolymer of 4-methyl-1-pentene and an α-olefin other than 4-methyl-1-pentene. The 4-methyl-1-pentene copolymer preferably has a ratio of structural units derived from 4-methyl-1-pentene of 80 mol % to 99 mol % of all structural units.

[0062] The other α-olefin in these copolymers may be a linear olefin, a branched olefin, or a cyclic olefin.

[0063] The other linear α-olefin is more preferably an α-olefin having from 2 to 20 carbon atoms, and even more preferably an α-olefin having from 2 to 3 carbon atoms. Examples of the other linear α-olefin include ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, 1-decene, 1-undecene, 1-dodecene, 1-tetradecene, 1-hexadecene, 1-octadecene, and 1-eicosene. Of these, ethylene, propylene, 1-butene, 1-pentene, 1-hexene, 1-octene, and 1-decene are preferred, ethylene and propylene are more preferred, and propylene is even more preferred.

[0064] The resin composition may be a mixture of a 4-methyl-1-pentene copolymer and another α-olefin copolymer. From the viewpoint of improving adhesion to the bag member and transparency, the resin composition preferably contains a 4-methyl-1-pentene copolymer and a butene copolymer.

[0065] The resin composition preferably has a total light transmittance of 85% or more, more preferably 90% to 100%, and even more preferably 92% to 100%, when measured on a 1 mm thick pressed sheet in accordance with JIS K7361-1:1997. By using a resin composition capable of producing a transparent molded body, the transparency of the port member 200 can be increased, thereby improving the visibility of the cell container. (Conditions for measuring total light transmittance) Apparatus: Fully automatic haze meter (manufactured by Tokyo Denshoku Co., Ltd., TC-HIII DPK) Light source: 12V 50W halogen lamp C light (lifespan 2000 hours) (manufactured by NARVA, 55928)

[0066] The resin composition preferably has a peel strength of 20 N / 15 mm or more and 100 N / 15 mm or less, more preferably 25 N / 15 mm or more and 100 N / 15 mm or less, and even more preferably 30 N / 15 mm or more and 100 N / 15 mm or less, as measured by a 180° peel test under the following measurement conditions. The higher the peel strength, the more likely it is that the port member 200 will be welded to the resin films 112 and 114, thereby improving the airtightness of the cell container 100. (Measurement Conditions) A 65 mm x 65 mm x 1.0 mm press sheet obtained from the resin composition is heat-sealed to the planar surface of a 65 mm x 65 mm x 0.3 mm substrate formed from a 4-methyl-1-pentene copolymer consisting of 4-methyl-1-pentene structural units and propylene structural units at a temperature of 240°C, a pressure of 0.1 MPa, a pressurization time of 5 seconds, and a seal width of 10 mm. The multilayer film is welded so that the weld line is perpendicular to the machine direction of the multilayer film. Then, a 180° peel test is performed at 23°C using a tensile tester. The peel strength is measured for three test pieces and the average value is calculated.

[0067] The resin composition preferably has a durometer type D hardness of 30 or more and 80 or less, more preferably 40 or more and 80 or less, and even more preferably 50 or more and 75 or less, as measured on a pressed sheet in accordance with ASTM D2240. By using a resin composition that can produce a molded product with high hardness, even when a port member 200 having a thin-walled portion such as a groove or a screw in the thin tubular portion 212 is produced, deformation during welding is less likely to occur, and when a soft tube is fitted, leakage of the internal fluid through a gap with the fitting portion can be prevented.

[0068] The resin composition preferably has a flexural modulus measured in accordance with ASTM D790 of 100 MPa to 2000 MPa, more preferably 200 MPa to 1700 MPa, and even more preferably 300 MPa to 1700 MPa. The lower the flexural modulus, the less likely the port member 200 is to be damaged when subjected to an impact, such as by being dropped. The higher the flexural modulus, the easier it is to insert a cap, an injection port, a tube, or the like, into the thin cylindrical portion 212.

[0069] 1-3. Thermal Adhesive Film The port member 200 may be joined to the bag member 110 by being directly welded to the resin films 112 and 114 as shown in FIGS. 1A and 1B, or may be joined to the bag member 110 via a thermal adhesive film 300 as shown in FIGS. 2A and 2B.

[0070] The thermal adhesive film 300 is disposed at a portion of the port member 200 joined to the heat-sealed joint 116 of the resin films 112 and 114, so as to surround the hollow port member 200 in the circumferential direction. The thermal adhesive film 300 is a single-layer or multi-layer resin sheet. The thermal adhesive film 300 adheres to the port member 200 before heat sealing to prevent misalignment of the port member 200 during heat sealing and to prevent a decrease in the airtightness of the cell container 100 due to the occurrence of gaps that are not heat-sealed due to the misalignment. Furthermore, the thermal adhesive film 300 is welded to the port member 200 on one side (the inner surface) and to the resin films 112 and 114 on the other side (the outer surface), thereby tightly adhering the port member 200 and these resin films without any gaps, thereby preventing a decrease in the airtightness of the cell container 100 due to the occurrence of gaps between them.

[0071] Fig. 4A is an exemplary cross-sectional view of a single-layer thermal adhesive film 300, Fig. 4B is an exemplary cross-sectional view of a two-layer laminate thermal adhesive film 300, and Fig. 4C is an exemplary cross-sectional view of a three-layer laminate thermal adhesive film 300. The thermal adhesive film 300 shown in Fig. 4A is made of a single layer of elastomer layer 310. Note that the layer structure of the laminate thermal adhesive film 300 is not limited to this, and may be four or more layers.

[0072] As shown in FIGS. 4A and 4B , the thermal adhesive film 300 preferably has an elastomer layer 310 containing a styrene-based elastomer (SEBS, SEPS, or SEEPS) on the side that comes into direct contact with the surface of the port member 200. The elastomer layer 310 is slightly adhesive and adheres to the port member 200 when it is joined to the resin film 112 or 114 (before heat sealing). This allows the thermal adhesive film 300 to adhere tightly to the port member 200, making it less likely for the port member 200 to shift position and reducing the likelihood of gaps forming between them, thereby improving the airtightness of the cell container 100. To more significantly enhance the effect of improving the airtightness, the elastomer layer 310 is preferably disposed so as to surround the entire periphery of the port member 200, and more preferably is welded to the entire periphery of the port member 200.

[0073] The styrene-based elastomer may be any elastomer containing styrene-derived structural units. The styrene-based elastomer may have a styrene-derived structural unit content of 5% by mass to 50% by mass, preferably 10% by mass to 40% by mass, and more preferably 10% by mass to 35% by mass. The styrene-based elastomer is preferably a hydrogenated product in which some or all of the unsaturated bonds derived from the raw material monomer, such as alkadiene (usually excluding unsaturated bonds in the benzene ring derived from styrene), have been converted to saturated bonds by hydrogenation.

[0074] Examples of the styrene-based elastomer include a block copolymer having a polystyrene block and a polyolefin block composed of structural units derived from an alkadiene having from 4 to 10 carbon atoms, and hydrogenated products thereof. The polyolefin block preferably contains structural units derived from isoprene or butadiene. The block copolymer may be a diblock copolymer composed of a polystyrene block and a polyolefin block bonded thereto, or a triblock copolymer composed of polystyrene blocks located at both ends and a polyolefin block located therebetween. The content of the polyolefin block relative to the total mass of the block copolymer may be from 5 to 95% by mass, preferably from 10 to 90% by mass, and more preferably from 20 to 90% by mass.

[0075] Specific examples of the styrene-based elastomer that is a block copolymer include block copolymers (SBCs) having a polystyrene block that forms the hard segment and a conjugated diene monomer block such as isoprene or butadiene that forms the soft segment, hydrogenated styrene-butadiene-styrene block copolymers (HSBRs), styrene-ethylene-propylene-styrene block copolymers (SEPSs), styrene-ethylene-butene-styrene block copolymers (SEBSs), and styrene-ethylene-ethylene-propylene block copolymers (SEEPSs). These styrene-based elastomers may be used alone or in combination of two or more.

[0076] Furthermore, from the viewpoint of facilitating adhesion of these elastomers to the surface of the port member 200 and enhancing reworkability for fine adjustment of the bonding position after bonding to the port member 200, the surface hardness measured by durometer hardness (type A) is preferably 40 or more and 95 or less, more preferably 50 or more and 90 or less, and even more preferably 60 or more and 85 or less.

[0077] The thermoplastic resin layer 320 disposed on the resin film 112 or resin film 114 side of the elastomer layer 310 can be a layer (sealant layer) provided to increase the welding strength of the thermal adhesive film 300 to the resin film 112 or resin film 114. Furthermore, as shown in Figure 4C, the elastomer layer 330 that may be disposed on the port member 200 side of the elastomer layer 310 can be a layer provided to increase the adhesion of the thermal adhesive film 300 to the port member 200 and the welding strength.

[0078] For example, when the material of the port member 200 is a relatively hard resin such as high-density polyethylene (HDPE), it is preferable to dispose the elastomer layer 330 to increase the welding strength of the thermal adhesive film 300 to the port member 200. When the material of the port member 200 includes polyethylene, the elastomer layer 330 preferably includes an ethylene-based elastomer. Specific examples of the ethylene-based elastomer of the elastomer layer 330 include ethylene-propylene copolymers, ethylene-propylene-butene copolymers, ethylene-octene copolymers, ethylene-butene copolymers, and ethylene-hexene copolymers.

[0079] Alternatively, when the material of the port member 200 includes polypropylene, the elastomer layer 330 preferably includes a propylene-based elastomer. Specific examples of the propylene-based elastomer of the elastomer layer 330 include propylene-ethylene copolymer, propylene-butene copolymer, propylene-ethylene-butene copolymer, etc.

[0080] From the viewpoint of improving the heat sealability with the resin film 112 or the resin film 114, the thermoplastic resin layer 320 preferably contains a 4-methyl-1-pentene copolymer.

[0081] The 4-methyl-1-pentene copolymer is preferably a copolymer of 4-methyl-1-pentene and an α-olefin other than 4-methyl-1-pentene, and is preferably a copolymer of 4-methyl-1-pentene and propylene. The molar ratio of 4-methyl-1-pentene to propylene (4-methyl-1-pentene / propylene) is preferably 45 / 55 or more and 95 / 5 or less, more preferably 70 / 30 or more and 93 / 7 or less, and even more preferably 75 / 25 or more and 90 / 10 or less.

[0082] The content of the 4-methyl-1-pentene copolymer contained in the thermoplastic resin layer 320 is preferably 75% by mass or more and 100% by mass or less, more preferably 80% by mass or more and 100% by mass or less, and even more preferably 85% by mass or more and 100% by mass or less, relative to the total mass of the thermoplastic resin layer 320.

[0083] The content of the ethylene-based copolymer contained in the elastomer layer 330 is preferably 50% by mass or more and 100% by mass or less, more preferably 60% by mass or more and 100% by mass or less, and even more preferably 70% by mass or more and 100% by mass or less, relative to the total mass of the elastomer layer 330.

[0084] When the thermal adhesive film has the structure shown in Fig. 4A or 4B, the elastomer layer 310 is preferably made of a styrene-based elastomer alone in order to ensure welding strength. Furthermore, when the thermal adhesive film has the structure shown in Fig. 4C, it is preferable that the elastomer layer 310 contains both a styrene-based elastomer and a butene-based copolymer in order to increase welding strength.

[0085] When the thermal adhesive film has a three-layer configuration as shown in Figure 4C, the content of the styrene-based elastomer contained in the elastomer layer 310 is preferably 20% by mass or more and 100% by mass or less, more preferably 40% by mass or more and 100% by mass or less, and even more preferably 50% by mass or more and 100% by mass or less, relative to the total mass of the elastomer layer 310.

[0086] The ratio of the content of the styrene elastomer to the butene copolymer (styrene elastomer / butene copolymer) in the elastomer layer 310 is preferably 20 / 80 or more and 100 / 0 or less, more preferably 30 / 70 or more and 90 / 10 or less, and even more preferably 40 / 60 or more and 70 / 30 or less.

[0087] The thickness of elastomer layer 310 is preferably 3 μm to 50 μm, more preferably 5 μm to 30 μm, and even more preferably 10 μm to 20 μm. If the thickness of elastomer layer 310 is within this range, when a heat sealing method is applied, high welding strength is maintained, and the elastomer layer 310 that protrudes from the portion pressed by the heat seal bar is less likely to produce burrs, thereby reducing the risk of the burrs falling off and contaminating the contents of the container.

[0088] The thickness of the thermoplastic resin layer 320 is preferably 5 μm or more and 100 μm or less, more preferably 10 μm or more and 50 μm or less, and even more preferably 20 μm or more and 30 μm or less. The thicker the thermoplastic resin layer 320, the higher the sealing property and welding strength of the thermal adhesive film 300 to the resin film can be. The thinner the thermoplastic resin layer 320, the less likely burrs can be generated.

[0089] The thickness of the elastomer layer 330 is preferably 3 μm or more and 100 μm or less, more preferably 5 μm or more and 50 μm or less, and even more preferably 10 μm or more and 30 μm or less. The thinner the elastomer layer 330, the less likely it is that burrs will occur. From the viewpoints of suppressing burrs during heat sealing and ensuring welding strength, the thickness of the elastomer layer 330 is preferably thinner than the thickness of the thermoplastic resin layer 320.

[0090] The thickness of the thermoplastic resin layer 320 is preferably ½ to 1 / 0.5 of the thickness of the elastomer layer 310 .

[0091] The thickness of the elastomer layer 330 is preferably ½ to 1 / 0.5 of the thickness of the elastomer layer 310 .

[0092] The total thickness of the thermal adhesive film 300 is preferably 10 μm to 100 μm, more preferably 20 μm to 70 μm, and even more preferably 30 μm to 50 μm. The thicker the thermal adhesive film 300, the easier it is to handle when temporarily fixing it to the port. The thinner the thermal adhesive film 300, the less likely it is to produce burrs during heat sealing.

[0093] In order to improve the airtightness of the port member 200, the heat sealing strength of the thermal adhesive film 300 when heat sealed to HDPE is preferably 10 N / 15 mm width or more and 300 N / 15 mm width or less.

[0094] The heat seal strength when heat sealed to HDPE can be measured by the following method. The thermal adhesive film 300 is cut into a size of 10 mm wide x 50 mm long to prepare a test piece. The elastomer layer 310 or 330 of the prepared test piece is placed on a high-density polyethylene plate (thickness 1 mm, melting point 130°C, density 960 kg / m). ３ The thermoplastic resin layer 320 of the test piece and the resin film 112 containing a 4-methyl-1-pentene copolymer are placed face to face and in contact with each other, and then the thermoplastic resin layer 320 of the test piece and the resin film 112 containing a 4-methyl-1-pentene copolymer are placed face to face and overlapped with each other, and then the test piece is welded (heat sealed) using a heat seal tester (thermal gradient heat seal tester TP-701-G, manufactured by Tester Sangyo Co., Ltd.) under the conditions of an upper temperature (heat seal temperature) of 220°C, a lower temperature of 23°C, a seal width of 10 mm, a seal pressure of 0.1 MPa, and a seal time of 5 seconds.

[0095] Next, the welded resin film 112, the thermal adhesive film 113, and the high-density polyethylene plate were removed from the testing machine, and 15 mm-wide strip-shaped test pieces were cut out in directions perpendicular and parallel to the weld line. The unwelded ends of the 15 mm-wide welded resin film and the high-density polyethylene plate were gripped with a 50 mm chuck distance using a tensile testing machine (Orientec Co., Ltd., Tensilon Universal Testing Machine, Model RTG-1250) and pulled 180° relative to the heat-sealed surface at a tensile speed of 300 mm / min and a temperature of 23°C, and the resin film was peeled off, and the maximum peel strength was measured. This maximum value was taken as the heat-seal strength (unit: N / 15 mm width). The heat-seal strength was measured for three test pieces, and the average value was calculated.

[0096] In order to improve the airtightness of the port member 200, the heat sealing strength of the thermal adhesive film 300 when heat sealed to hPP is preferably 10 N / 15 mm width or more and 300 N / 15 mm width or less.

[0097] The heat seal strength when heat-sealed to hPP can be measured by the following method. A thermal adhesive film 300 is cut into a size of 10 mm wide x 50 mm long to prepare a test piece. The elastomer layer 310 or elastomer layer 330 of the prepared test piece is brought into contact with an hPP plate (thickness 1 mm, melting point 160°C, MFR 230°C, 2.16 kgf = 9 g / 10 min) so that they face each other, and then the thermoplastic resin layer 320 of the test piece is placed face-to-face with a resin film 112 containing a 4-methyl-1-pentene copolymer, which is then overlapped. Then, using a heat seal tester (thermal gradient heat seal tester TP-701-G, manufactured by Tester Sangyo Co., Ltd.), the test pieces are welded (heat sealed) under the following conditions: upper temperature (heat seal temperature) 220°C, lower temperature 23°C, seal width 10 mm, seal pressure 0.1 MPa, and seal time 5 seconds.

[0098] Next, the welded resin film 112, thermal adhesive film 113, and hPP plate were removed from the testing machine, and 15 mm-wide strip test pieces were cut out in directions perpendicular and parallel to the weld line. The unwelded ends of the 15 mm-wide welded resin film and hPP plate were gripped with a 50 mm chuck distance using a tensile testing machine (Orientec Co., Ltd., Tensilon Universal Material Testing Machine, Model RTG-1250) and pulled 180° relative to the heat-sealed surface at a tensile speed of 300 mm / min and a temperature of 23°C, allowing the resin film to peel off. The maximum peel strength was measured. This maximum value was taken as the heat-seal strength (unit: N / 15 mm width). The heat-seal strength was measured for three test pieces, and the average value was calculated.

[0099] The width of the thermal adhesive film 300 in the width direction of the joint 116 (the direction perpendicular to the tangent line contacting the outer periphery of the bag member 110 at the position where the thermal adhesive film 300 is arranged) may be the same as the width of the joint 116 at the same position ( FIG. 5A ), may be smaller than the width of the joint 116 ( FIG. 5B ), or may be larger than the width of the joint 116 ( FIG. 5C ). According to the findings of the inventors, the material of the thermal adhesive film 300 described above is not easily eluted in the culture solution. Therefore, even if the thermal adhesive film 300 extends further inward than the joint 116 in the bag member 110, contamination of the culture solution does not occur.

[0100] From the viewpoint of ensuring airtightness of the culture medium, the bonding width L (heat seal width) of the thermal adhesive film 300 in the width direction of the joint 116 (the direction in which the port member 200 connects the inside and outside of the bag member 110) is preferably 1 mm or more and 50 mm or less, more preferably 5 mm or more and 20 mm or less, and even more preferably 5 mm or more and 10 mm or less.

[0101] The thermal adhesive film may contain additives such as heat stabilizers, anti-slip agents, anti-blocking agents, lubricants, pigments, dyes, plasticizers, hydrochloric acid absorbers, antioxidants, crystal nucleating agents, anti-fungal agents, fillers (inorganic fillers, organic fillers), and softeners.

[0102] 1-4. Manufacturing Method of Cell Container The cell container 100 can be produced by overlapping two resin films 112 and 114, or by folding one resin film 112 and overlapping the ends, and then heat-sealing the ends to form a bag.

[0103] When the resin films are overlapped and arranged, the port member 200 is sandwiched between the ends of the overlapping resin films. At this time, the port member 200 is arranged so that one end of the hollow port member 200 is located outside the resin film and the other end is located inside the heat-sealed region of the resin film. At this time, a thermal adhesive film 300 may be arranged between the port member 200 and the resin film. The thermal adhesive film 300 may be arranged so as to surround the entire circumference of the hollow port member 200 (it may be wrapped around the port member 200), or two thermal adhesive films 300 may be arranged above and below the port member 200 so as to be in contact with the respective resin films. However, it is preferable to arrange the thermal adhesive film 300 so as to surround the entire circumference of the port member 200.

[0104] The thermal adhesive film 300 may have a protective sheet placed on one or both sides before being used to manufacture the cell container 100. At this time, the protective sheet is peeled off, and the thermal adhesive film 300 is placed between the port member 200 and the resin film.

[0105] Heat sealing should be performed at a sealing temperature above the melting point of the resin that is the material of the port member 200 if the resin is a crystalline resin, or above the glass transition temperature of the resin if the resin is an amorphous resin, and above the melting point of the heat seal layer of the resin film.Taking into consideration the speed at which heat is transferred from the heat seal bar to the welding interface, which depends on the thickness of the resin sheet, productivity, and heat seal strength, heat sealing can be performed at a temperature of, for example, 100°C to 240°C, preferably 100°C to 200°C, at a pressure of 0.1 MPa to 1 MPa, preferably 0.1 MPa to 0.5 MPa, for 1 second to 10 seconds, preferably 1 second to 5 seconds.

[0106] The resin film and the port member 200 are joined by the heat sealing, or the resin film and the port member 200 are joined via the thermal adhesive film 300. When the port member 200 is a molded article of a resin composition containing a 4-methyl-1-pentene copolymer, the port member 200 is well fused to the heat seal layer of the resin film, thereby sufficiently increasing adhesion, and therefore the port member 200 and the resin film may be joined directly. When the port member 200 is a molded article of a resin composition containing polyethylene or polypropylene, joining via the thermal adhesive film 300 can increase adhesion between the port member 200 and the bag member 110 formed from the resin film.

[0107] When the port member 200 is a molded article made of a resin composition containing a 4-methyl-1-pentene copolymer, the port member 200 is in close contact with the resin film, making it less likely for the port member 200 to shift position during heat sealing. Therefore, the loss of airtightness of the cell container 100 due to gaps that are not heat-sealed due to the above-mentioned shift in position is less likely to occur.

[0108] When the thermal adhesive film 300 is used, the thermal adhesive film 300 is in close contact with the port member 200, so that misalignment of the thermal adhesive film 300 during heat sealing is unlikely to occur. Therefore, the loss of airtightness of the cell container 100 due to gaps not being heat-sealed caused by the misalignment is unlikely to occur. Furthermore, depending on the number of layers of the thermal adhesive film 300, the elastomer layer 310 or the elastomer layer 330 is welded to the port member 200, and the thermoplastic resin layer 320 is welded to the resin film, so that the loss of airtightness of the cell container 100 due to gaps being formed between the thermal adhesive film 300 and the port member 200 or the resin film is unlikely to occur.

[0109] When overlapping and arranging the resin films, the resin films may be shaped into a predetermined shape by methods such as vacuum forming, pressure forming, and vacuum / pressure forming. In this case, the edges of the resin films are required to be overlapped. In addition to the method of sandwiching the overlapped resin films between hot plates for heat sealing, impulse sealing, high-frequency welding, and laser welding can also be applied as methods for joining the resin films. For example, in the case of laser welding, as described in Japanese Patent No. 4279674, a method can be used in which the overlapping resin films are irradiated with laser light having a wavelength of 1.8 to 2 μm emitted from a Ho-YAG laser or a Tm fiber laser, or with laser light having a wavelength of 10.6 μm emitted from a carbon dioxide laser, to the joined portions of the overlapping resin films, causing the resin films to directly absorb the energy of the laser light and melt the joined portions, thereby welding them.

[0110] Furthermore, the number of resin films used to fabricate the cell container 100 is not limited to two. A single resin film may be folded and arranged with its ends overlapping, and the ends may be heat-sealed to form a bag, or a cell container may be formed using three or more resin films.

[0111] Furthermore, after the cell container is produced, it can be sterilized before use. As a sterilization method, gamma ray sterilization is preferred because the film material is resistant to gamma ray irradiation. The gamma ray irradiation dose is preferably in the range of 10 to 50 kGy. Furthermore, from the viewpoint of preventing contamination and maintaining sterility, it is preferred that the cell container be used as a single-use item.

[0112] 2. Method for Culturing Cells The cell container described above can be used for culturing various types of cells.

[0113] Specifically, an injection port for injecting a drug or the like and a tube for connecting to a Luer port are attached to each port member 200. Then, a culture solution containing cells (cell suspension) is introduced into the cell container 100 from the Luer port connected to the tube. Note that a culture solution not containing cells may be introduced into the cell container 100 first, and then the cell suspension may be introduced.

[0114] Thereafter, the cell container 100 is placed in an incubator to culture the cells. The conditions for culturing the cells are not particularly limited and may be selected depending on the cells to be cultured.

[0115] During the culture, a drug may be injected through the injection port, or a portion of the cells may be sampled to check the culture state. Also, some or all of the cells may be transferred to another cell container 100 via a tube attached to the port member.

[0116] Finally, the cultured cells are collected through a tube attached to the port member.

[0117] The cells targeted by the cell container 100 are not particularly limited and may be differentiated somatic cells or undifferentiated stem cells. The cells may be living cells or dead cells.

[0118] It should be noted that each of the above-described embodiments represents an example of the present invention, and the present invention is not limited to the above-described embodiments. It goes without saying that various other embodiments are possible within the scope of the concept of the present invention.

[0119] The present invention will be described in detail based on examples, but the present invention is not limited to these examples.

[0120] [Experiment 1] In Experiment 1, a test was conducted in which a port member or a press sheet of the material thereof was directly bonded to a multilayer film that was to be used as a bag member.

[0121] 1. Preparation of multilayer and single layer films Multilayer films having the following layer configuration were prepared. For layers prepared using multiple materials, the blending amounts of the materials used in preparing each layer are shown in parentheses.

[0122] Multilayer film 1-1: surface layer / core layer / heat seal layer = [4MP1-3 (81 parts by mass) + 4MP1-4 (10 parts by mass) + butene-1 (9 parts by mass)] / 4MP1-1 / [4MP1-2 (90 parts by mass) + butene-1 (10 parts by mass)] = 60 μm / 180 μm / 60 μm Multilayer film 1-2: surface layer / core layer / heat seal layer = [4MP1-2 (91 parts by mass) + butene-2 ​​(9 parts by mass) + ethylene / silicone block copolymer (1 part by mass)] / 4MP1-1 / same as surface layer = 30 μm / 40 μm / 30 μm Multilayer film 1-3: surface layer / core layer / heat seal layer = [4MP1-2 (40 parts by mass) + 4MP1-3 (60 parts by mass) + ethylene / silicone block copolymer (1 part by mass) / SEBS / same as surface layer] = 75 μm / 150 μm / 75 μm Multilayer film 1-4: surface layer / core layer / heat seal layer = [4MP1-2 (40 parts by mass) + 4MP1-3 (60 parts by mass) + ethylene / silicone block copolymer (1 part by mass) / SEBS / same as surface layer] = 25 μm / 50 μm / 25 μm Multilayer film 1-5: surface layer / core layer / heat seal layer = [4MP1-2 (81 parts by mass) + 4MP1-4 (10 parts by mass) + butene-1 (9 parts by mass) / 4MP1-1 / 4MP1-2 (90 parts by mass) + butene-1 (10 parts by mass)] = 60 μm / 180 μm / 60 μm Monolayer film 1-6: LLDPE monolayer film, thickness 300 μm Monolayer film 1-7: LLDPE monolayer film, thickness 100 μm

[0123] The raw materials for the multilayer film are as follows. (4-Methyl-1-pentene Copolymer 1) A 4-methyl-1-pentene copolymer (melting point: 224°C) having a proportion of 97.6 mol% of structural units derived from 4-methyl-1-pentene, a proportion of 1.44 mol% of structural units derived from 1-hexadecene, and a proportion of 0.96 mol% of structural units derived from 1-octadecene was designated 4MP1-1. (4-Methyl-1-pentene Copolymer 2) A 4-methyl-1-pentene copolymer (melting point: 130°C) having a proportion of 85 mol% of structural units derived from 4-methyl-1-pentene and a proportion of 15 mol% of structural units derived from propylene was designated 4MP1-2. (4-Methyl-1-pentene Copolymer 3) A 4-methyl-1-pentene copolymer having a proportion of 72 mol% of structural units derived from 4-methyl-1-pentene and a proportion of 28 mol% of structural units derived from propylene was designated 4MP1-3. (4-Methyl-1-pentene Copolymer 4) A 4-methyl-1-pentene copolymer (manufactured by Mitsui Chemicals, Inc., product: TPX, brand: MX004, melting point: 228°C) containing structural units derived from 4-methyl-1-pentene, structural units derived from 1-hexadecene, and structural units derived from 1-octadecene was designated 4MP1-4. (Butene Copolymer 1) A butene copolymer (manufactured by Mitsui Chemicals, Inc., product name: Tafmer, brand: BL2491M, melting point: 100°C) containing structural units derived from 1-butene and structural units derived from propylene was designated butene-1. (Butene-based copolymer 2) A butene-based copolymer containing structural units derived from 1-butene and structural units derived from ethylene (manufactured by Mitsui Chemicals, Inc., product name: Tafmer, brand name: BL3450M, melting point: 100°C) was used as butene-2. (Styrene-based elastomer) Kraton G1657VS (SEBS, styrene content: 13% by mass) manufactured by Kraton Polymer Japan Co., Ltd. was used as SEBS. (Ethylene / silicone block copolymer) A polyethylene (P'-1) having a vinyl group at one end, synthesized in accordance with the method described in Synthesis Example 2 of WO 2012 / 098865, was used as an ethylene / silicone block copolymer.(Linear Low-Density Polyethylene) Evolue SP2040 (melting point: 116°C) manufactured by Prime Polymer Co., Ltd. was used as LLDPE.

[0124] 2. Port Member 2-1. Preparation of Materials 4MP1-1 and 4MP1-2 were prepared as materials for the port member.

[0125] 2-2. Measurement of physical properties The physical properties of the materials prepared above were measured by the following methods.

[0126] 2-2-1. Total Light Transmittance Press sheets (65mm x 65mm x 1mm) were produced using a Shinto Metal Industries compression molding machine (maximum clamping force 10 tons) ASF-10 and a Shinto Metal Industries cooling compression molding machine (maximum clamping force 37 tons) NSF-37. The molding conditions for each material were as follows: 4MP1-1 Heating temperature: 270°C, preheating: 10 minutes, pressure: 3 minutes, cooling: 5 minutes 4MP1-2 Heating temperature: 200°C, preheating: 3 minutes, pressure: 2 minutes, cooling: 3 minutes

[0127] The total light transmittance of this press sheet was measured in accordance with JIS K7361-1:1997 under the following conditions. (Conditions for measuring total light transmittance) Apparatus: Fully automatic haze meter (TC-HIII DPK, manufactured by Tokyo Denshoku Co., Ltd.) Light source: 12V 50W halogen lamp C light (lifespan 2000 hours) (55928, manufactured by NARVA)

[0128] 2-2-2. Durometer Hardness (Type D) (Surface Hardness) Test pieces were prepared under the following conditions using a 1 mm thick resin sheet obtained by a compression molding machine similar to that used to measure the total light transmittance, and the surface hardness was measured. Test standard: ASTM D2240 Test temperature: 23°C Apparatus: Durometer Hardness (Type D) Hardness Tester The instantaneous value immediately after the indenter of the hardness tester started to contact was read, and this was taken as the surface hardness.

[0129] 2-2-3. Melt flow rate (MFR) Measured in accordance with ASTM D1238 (2013). Measurements were performed under the following conditions according to the melting point of each material: 4MP1-1: 260°C, 5 kgf 4MP1-2: 230°C, 2.16 kgf

[0130] 2-2-4. Flexural Modulus The flexural modulus of the above-mentioned 4MP1-1 was measured in accordance with ASTM D790.

[0131] 2-2-5. Density Measurement was carried out in accordance with JIS K7112-2:2023 using a resin sheet having a thickness of 1 mm obtained by a compression molding machine similar to that used in measuring the total light transmittance.

[0132] 2-2-6. Appearance of port section Port members with the shapes shown in Figure 3 were produced from each material by injection molding. The produced port members had a thin and thick cylindrical section, both of which had circular cross sections, with an outer diameter of 8 mm, an inner diameter of 4 mm, an outer diameter of 10 mm, and an inner diameter of 6 mm.

[0133] The appearance of the formed port member was visually inspected and judged based on the following criteria: transparent so that the opposite side could be easily seen, slightly colored and semi-transparent so that the opposite side was difficult to see, or opaque so that the opposite side could not be seen.

[0134] The measurement results for the port members are shown in Table 1.

[0135]

[0136] 3. Evaluation 3-1. Peel Strength The same press sheets (65 × 65 × 1 mm) as those used in the measurement of total light transmittance described above were used for heat sealing, and the peel strengths between Multilayer Films 1-1 to 1-5 and Single Layer Films 1-6 to 1-7 and the press sheets were determined.

[0137] The multilayer film was placed on top of the press sheet so that the sealant layer surface was in contact with the surface of the press sheet, and heat-sealed using a heat seal tester (Tester Sangyo Co., Ltd., thermal gradient heat seal tester, model number TP-701-G) under the following conditions: upper temperature of the heat seal bar (heat seal temperature) 240°C, lower temperature of the heat seal bar 23°C, seal width 10 mm, seal pressure 0.1 MPa, and seal time 5 seconds. The heat seal was performed so that the weld line was perpendicular to the MD direction of the multilayer film. The welded multilayer film and press sheet were then removed from the tester. Then, a 15 mm wide strip test piece was cut out perpendicular to the weld line direction. Using a tensile tester (Tensilon universal material testing machine, model RTG-1250, manufactured by Orientec Co., Ltd.), the welded strip-shaped test pieces were peeled at 180° under conditions of a test temperature of 23°C, a chuck distance of 50 mm, and a tensile speed of 300 mm / min, and the maximum peel strength was measured and taken as the peel strength (unit: N / 15 mm). The peel strength was measured for three test pieces and the average value was calculated.

[0138] The peeled test pieces were also visually inspected to evaluate how they peeled. When the test piece was torn at the edge of the sealed area (edge ​​tear), it was determined that the multilayer film and the press sheet were very well welded. When delamination occurred between the multilayer film layers (interlayer delamination), it was determined that the multilayer film and the press sheet were very well welded. On the other hand, when delamination occurred at the interface between the multilayer film and the press sheet (interface delamination), it was determined that the weldability between the multilayer film and the press sheet was poor.

[0139] Two sheets of film measuring 120 mm in length and 120 mm in width were prepared by cutting out each of multilayer films 1-1 to 1-4 and single layer films 1-5 to 1-6, and these were arranged facing each other. Three sides were heat-sealed at a sealing temperature of 160°C and a sealing pressure of 0.1 MPa to obtain bag members.

[0140] A port member was placed at the opening of the bag member, and heat-sealed with a welding line width (joint width) of 10 mm under conditions of a sealing temperature of 240°C, a sealing pressure of 0.1 MPa, and a sealing time of 4 seconds to obtain a cell culture bag with a port.

[0141] The resulting cell culture bag with port was filled with 50 ml of air, and the port member was sealed with a silicone resin inner stopper with an upper diameter of 13 mm, a lower diameter of 10 mm, and a height of 18 mm. After leaving the bag to stand for 30 minutes, the bag was submerged in water 30 cm deep (depth 31.5 cm, width 18.5 cm, height 24.4 cm), and visually inspected for air bubbles leaking outside the bag for 30 minutes. ○: No air bubbles were observed during the 30 minutes the bag was submerged in water. ×: Air bubbles were observed during the 30 minutes the bag was submerged in water.

[0142] The measurement results and evaluation results of the physical properties of each material are shown in Tables 2 and 3.

[0143]

[0144]

[0145] As shown in Tables 2 and 3, all of multilayer films 1 to 5 (all bag member materials) containing a 4-methyl-1-pentene copolymer in the heat seal layer were welded to the port member. Furthermore, as shown in Table 1, the port parts made from resin compositions containing a 4-methyl-1-pentene copolymer and having a total light transmittance of 85% or more when measured on a pressed sheet (thickness 1 mm) had excellent transparency and good visibility.

[0146] [Experiment 2] In Experiment 2, a test was conducted in which a port member was joined to a bag member formed of a multilayer film via a thermal adhesive sheet.

[0147] 1. Preparation of multilayer film The multilayer film 1-5 produced in Experiment 1 was used as the multilayer film 2-1 in Experiment 2. The oxygen permeability of the multilayer film 2-1 was 4.2 L / (m ２ · day · atm), carbon dioxide permeability is 15 L / (m ２ · day · atm), water vapor permeability is 10 g / m ２ ・It was day.

[0148] The multilayer film 1-2 produced in Experiment 1 was used as the multilayer film 2-2 in Experiment 2. The oxygen permeability of the multilayer film 2-2 was 12 L / (m ２ · day · atm), carbon dioxide permeability is 45 L / (m ２ · day · atm), water vapor permeability is 32 g / m ２ ・It was day.

[0149] The multilayer film 1-3 produced in Experiment 1 was used as the multilayer film 2-3 in Experiment 2. The oxygen permeability of the multilayer film 2-3 was 4.2 L / (m ２ · day · atm), carbon dioxide permeability is 15 L / (m ２ · day · atm), water vapor permeability is 11 g / m ２ ・It was day.

[0150] The multilayer film 1-4 produced in Experiment 1 was used as the multilayer film 2-4 in Experiment 2. The oxygen permeability of the multilayer film 2-4 was 12 L / (m ２ ·day·atm), carbon dioxide permeability is 44 L / (m ２ · day · atm), water vapor permeability is 31 g / m ２ ・It was day.

[0151] The oxygen permeability [L / (m ２ ・day・atm)], carbon dioxide permeability [L / (m ２ ·day·atm)], and moisture permeability [g / m ２ ·day] was measured by the following method.

[0152] The multilayer films 2-1 to 2-4 were cut into a shape of 30 mm width x 30 mm length and used as test pieces.

[0153] The oxygen permeability and carbon dioxide permeability were measured in accordance with JIS K 7126-1:2006 using a differential pressure gas permeability measuring device (manufactured by Toyo Seiki Seisakusho) at a test temperature of 23°C and a test humidity of 0% RH, with the measurement area of ​​the film containing 4-methyl-1-pentene copolymer being 5 cm. ２The measurement area of ​​the film containing a 4-methyl-1-pentene copolymer was adjusted by preparing two adhesive aluminum masks manufactured by Modern Control Co., Ltd., each with a 25 mm diameter hole in the center, and laminating the film containing the 4-methyl-1-pentene copolymer to be measured between the two masks so that the film was sandwiched between them. Specifically, the film containing a 4-methyl-1-pentene copolymer was positioned so that the central hole overlapped the two masks.

[0154] The moisture permeability was calculated in accordance with the isobaric method (cup-type gravimetric method) described in JIS Z 0208:2021 under condition B (test temperature 40°C, test humidity 90% RH).

[0155] 2. Preparation of thermal adhesive film 2-1. Preparation of materials The following materials were prepared. The density of each material was measured in accordance with JIS K7112-2:2023 (density gradient tube method).

[0156] 2-1-1. 4-Methyl-1-pentene copolymer 4MP1: A copolymer of 85 mol% 4-methyl-1-pentene and 15 mol% propylene

[0157] 2-1-2. Ethylene-based polymer PE-1: Mitsui Chemicals, Inc., Tafmer DF7350 (ethylene-1-butene copolymer, density 0.870 g / cm ３ , MFR 230°C, 2.16 kgf = 65 g / 10 min) PE-2: Tafmer A-4085S (ethylene-1-butene copolymer, density 0.885 g / cm), manufactured by Mitsui Chemicals, Inc. ３ , MFR 230°C, 2.16 kgf = 6.7 g / 10 min) PE-3: Tafmer A-20085S (ethylene-1-butene copolymer, density 0.885 g / cm), manufactured by Mitsui Chemicals, Inc. ３ , MFR 230°C, 2.16 kgf = 33 g / 10 min) PE-4: Mitsui Chemicals, Inc., Tafmer A-4090S (ethylene-1-butene copolymer, density 0.893 g / cm ３ , MFR230℃・2.16kgf=6.7g / 10min)

[0158] 2-1-3. Propylene-based polymers PP-1: Mitsui Chemicals, Inc., Tafmer XM-7070 (propylene-butene copolymer, melting point 75°C, MFR 230°C, 2.16 kgf = 7 g / 10 min)

[0159] 2-1-4. Butene polymer PB: Mitsui Chemicals, Inc., Tafmer BL2491M (butene-propylene copolymer, density 0.90 kg / cm ３ , MFR230℃・2.16kgf=9g / 10min)

[0160] 2-1-5. Styrene-based elastomer SEBS: Kraton G1657VS (styrene content: 13% by mass, density: 0.90 g / cm), manufactured by Kraton Polymer Japan Co., Ltd. ３ , MFR230℃・2.16kgf=9g / 10min)

[0161] 2-2. Preparation of Thermal Adhesive Film The above materials were appropriately selected and mixed (dry blended) to form the resin material for thermoplastic resin layer 1, the resin material for elastomer layer 1, and the resin material for elastomer layer 2. Of the three-layer thermal adhesive film to be produced, thermoplastic resin layer 1 is the layer disposed on the heat seal surface side of resin film 112 and resin film 114, and the layer disposed on the port member side is elastomer layer 2 (sealant layer).

[0162] Next, each resin material was charged into the hopper of a multilayer film molding machine equipped with a multilayer T-die (lip width 350 mm), two single-screw extruders (for thermoplastic resin layer 1 and elastomer layer 2), and one single-screw extruder (for elastomer layer 1). The cylinder temperatures of each extruder (feed section / compression section to metering section) were 140°C / 240°C for both the extruders for thermoplastic resin layer 1 and elastomer layer 2, and 170°C / 250°C for the extruder for the elastomer layer. The die temperature was 240°C. Melt-kneaded mixtures of the resin material for thermoplastic resin layer 1, the resin material for elastomer layer 1, and the resin material for elastomer layer 2 were co-extruded from the T-die to have the cross-sectional layer configuration and total thickness shown in Table 1, and cast-molded using a chill roll with a mirror surface set at 20°C. The resulting mixtures were then taken up to obtain thermal adhesive films (total thickness: 50 μm to 100 μm).

[0163] Tables 4 and 5 show the blending ratio of the resin material of each layer and the thickness configuration of the produced thermal adhesive films 1 to 6.

[0164]

[0165]

[0166] 3. Fabrication of port member High density polyethylene (melting point 130°C, density 960 kg / m ３ A hollow cylindrical port member 1 having a height of 30 mm, an inner diameter of 12 mm, and an outer diameter of 15 mm was produced by injection molding using a resin having a melting point of 1.5 mm and a melting point of 1.5 mm.

[0167] A hollow cylindrical port member 2 having a height of 30 mm, an inner diameter of 12 mm, and an outer diameter of 15 mm was produced by injection molding using polypropylene resin (melting point 160° C., MFR 230° C., 2.16 kgf=9.0 g / 10 min).

[0168] 4. Evaluation 4-1. Temporary Fixation Test The obtained thermal adhesive films 1 to 6 were cut into strips. The thermal adhesive films were arranged so that the thermoplastic resin layer 2 side of the strip-shaped thermal adhesive films faced the port member, and the thermal adhesive films were temporarily fixed so as to surround the entire periphery of the port member, and the ease of temporary fixing of the thermal adhesive films to the port member before heat welding was evaluated according to the following criteria: ○: The thermal adhesive film adhered to the entire periphery of the port member and did not peel off from the port member ×: The thermal adhesive film sheet did not adhere to the port member and temporary fixing was not possible

[0169] 4-2. Heat sealing test The obtained thermal adhesive films 1 to 6 were cut into a size of 10 mm in width and 50 mm in length to prepare test pieces. The elastomer layer 2 of the prepared test piece was placed between a high-density polyethylene plate (thickness 1 mm, melting point 130°C, density 960 kg / m ３The test pieces were placed face-to-face in contact with a polyethylene plate (thickness 1 mm, melting point 160°C, MFR 230°C, 2.16 kgf = 13 g / 10 min) or a propylene plate (thickness 1 mm, melting point 160°C, MFR 230°C, 2.16 kgf = 9.0 g / 10 min), and the thermoplastic resin layer 1 of the test piece and the multilayer film 2-1 were placed face-to-face and overlapped, and then heat-sealed using a heat-sealing tester (thermal gradient heat-sealing tester TP-701-G, manufactured by Tester Sangyo Co., Ltd.) under the conditions of an upper temperature (heat-sealing temperature) of 220°C, a lower temperature of 23°C, a seal width of 10 mm, a seal pressure of 0.1 MPa, and a seal time of 5 seconds.

[0170] Next, the resin film, heat-bonded film, and high-density polyethylene or polypropylene plate integrated by welding were removed from the testing machine and cut into 15 mm wide strip test pieces in a direction perpendicular to the welding line and parallel to it. These 15 mm wide welded test pieces were then pulled and peeled at a 180° angle relative to the heat-sealed surface using a tensile testing machine (Orientec Co., Ltd., Tensilon Universal Testing Machine, Model RTG-1250) under conditions of a chuck distance of 50 mm, a pulling speed of 300 mm / min, and a temperature of 23°C, and the maximum peel strength was measured. This maximum value was taken as the heat-seal strength (unit: N / 15 mm). The heat-seal strength was measured for three test pieces and the average value was calculated.

[0171] Two films measuring 100 mm in length and 100 mm in width were prepared by cutting out multilayer films 2-1 to 2-4, and placed opposite each other. Three sides were heat-sealed at a sealing temperature of 160°C and a sealing pressure of 0.1 MPa to obtain a bag member.

[0172] The obtained thermal adhesive films 1 to 6 were each cut into a 10 mm wide strip, and the elastomer layer 2 of the prepared test piece was placed opposite port member 1 or 2, brought into contact with the entire outer periphery of the port, and the thermal adhesive film was temporarily fixed to the port member. The port member with the thermal adhesive film temporarily fixed thereto was placed at the opening of the bag member and heat-sealed with a welding line width (joint width) of 10 mm under conditions of a sealing temperature of 220°C, a sealing pressure of 0.1 MPa, and a sealing time of 5 seconds, to obtain a cell culture bag with a port.

[0173] The resulting cell culture bag with port was filled with 50 ml of air, and the port member was sealed with a silicone resin inner stopper with an upper diameter of 13 mm, a lower diameter of 10 mm, and a height of 18 mm. After leaving the bag to stand for 30 minutes, the bag was submerged in water 30 cm deep (depth 31.5 cm, width 18.5 cm, height 24.4 cm), and visually inspected for air bubbles leaking outside the bag for 30 minutes. ○: No air bubbles were observed during the 30 minutes the bag was submerged in water. ×: Air bubbles were observed during the 30 minutes the bag was submerged in water.

[0174] The evaluation results of Thermal Adhesive Films 1 to 6 are shown in Tables 6 and 7. Table 6 shows the results of a temporary fixing test using a high-density polyethylene port member 1, a heat sealing test using a high-density polyethylene plate, and an airtightness test using a high-density polyethylene port member 1. Table 7 shows the results of a temporary fixing test using a high-density polypropylene port member 2, a heat sealing test using a polypropylene plate, and an airtightness test using a polypropylene port member 2.

[0175]

[0176]

[0177] As shown in Tables 6 and 7, all of multilayer films 2-1 to 2-4 (all bag member materials) containing a 4-methyl-1-pentene copolymer in the heat seal layer were welded to the port member. Furthermore, as is clear from Tables 6 and 7, by joining a port member to a bag member with high oxygen permeability via a single-layer or multilayer heat-adhesive film having an elastomer layer containing a styrene-based elastomer, the port member was temporarily fixed, and positional displacement during heat sealing was suppressed.

[0178] [Experiment 3] In Experiment 3, a cell culture test was carried out.

[0179] 3-1. Fabrication of cell culture bag with port 3-1-1. Cell culture bag 1 Two sheets of film measuring 120 mm in length and 120 mm in width were prepared by cutting out the multilayer film 1-2 fabricated in Experiment 1, and were arranged facing each other. Three sides were heat-sealed to a width of 10 mm at a sealing temperature of 160°C and a sealing pressure of 0.1 MPa to obtain a bag member.

[0180] The port member made from 4MP1-1 in Experiment 1 was placed at the opening of the bag member, and heat-sealed at a sealing temperature of 240°C, a sealing pressure of 0.1 MPa, and a sealing time of 4 seconds with a welding line width (joint width) of 10 mm to obtain a cell culture bag 1 with a port. The area of ​​the culture space of the cell culture bag 1 with a port was 100 cm. ２ 3-1-2. Cell culture bag 2 Two films each measuring 120 mm in length and 120 mm in width were prepared by cutting out the single layer film 1-7 produced in Experiment 1, and were arranged facing each other. Three sides were heat-sealed at a sealing temperature of 160°C and a sealing pressure of 0.1 MPa to a width of 10 mm to obtain a bag member.

[0181] The port member 1 made from high-density polyethylene in Experiment 2 was placed at the opening of the bag member, and heat-sealed at a sealing temperature of 220°C, a sealing pressure of 0.1 MPa, and a sealing time of 4 seconds with a welding line width (joint width) of 10 mm to obtain a cell culture bag 2 with a port. The area of ​​the culture space of the cell culture bag 2 with a port was 100 cm. ２ It was.

[0182] 3-1-3. Cell culture bag 3 Two sheets of multilayer film 1-2 produced in Experiment 1, each measuring 100 mm in length and 50 mm in width, were cut out and placed facing each other. Three sides were heat-sealed at a sealing temperature of 160°C and a sealing pressure of 0.1 MPa to obtain a bag member.

[0183] The port member made from 4MP1-1 in Experiment 1 was placed at the opening of the bag member, and heat-sealed at a sealing temperature of 240°C, a sealing pressure of 0.1 MPa, and a sealing time of 4 seconds with a welding line width (joint width) of 10 mm to obtain a cell culture bag 3 with a port. The area of ​​the culture space of the cell culture bag 3 with a port was 24 cm. ２ It was.

[0184] 3-1-4. Cell culture bag 4 Two films each measuring 100 mm in length and 50 mm in width were prepared by cutting out the single-layer film 1-7 produced in Experiment 1, and were placed facing each other. Three sides were heat-sealed at a sealing temperature of 160°C and a sealing pressure of 0.1 MPa to a width of 10 mm to obtain a bag member.

[0185] The port member 1 made from high-density polyethylene in Experiment 2 was placed at the opening of the bag member, and heat-sealed at a sealing temperature of 220°C, a sealing pressure of 0.1 MPa, and a sealing time of 4 seconds with a welding line width (joint width) of 10 mm to obtain a cell culture bag 4 with a port. The area of ​​the culture space of the cell culture bag 4 with a port was 24 cm. ２ It was.

[0186] The cell culture bag with the port was sterilized by gamma irradiation at a dose of 25 kGy. The Sterility Assurance Level (SAL) of the medical device measured in accordance with BS EN556-1:2001 for the cell culture bag manufactured under the same conditions and sterilized by gamma irradiation at a dose of 25 kGy was 10. －６ It was.

[0187] 3-2. Cell culture test 3-2-1. Test 1 2 x 10 ７Human chronic myeloid leukemia cell line K562 (RIKEN BRC, RCB0027) was suspended in 100 ml of RPMI 1640 medium (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% FBS (BioWest), 1% GlutaMAX™-I supplement (Thermo Fisher Scientific), and 1% penicillin / streptomycin (Fujifilm Wako Pure Chemical Industries, Ltd.). The cells were introduced into the sterilized ported cell culture bag 1 and cell culture bag 2 and incubated at 37°C and 5% CO. ２ CO maintained at ２ The cells were left to stand in an incubator for culturing, and the cell suspension was collected from the port using a syringe 3 and 7 days after the start of culturing.

[0188] The cell density and cell viability of the cell suspension were calculated using the trypan blue exclusion method 3 and 7 days after the start of culture. Specifically, the cell suspension was mixed with 0.4 w / v% trypan blue solution (Fujifilm Wako Pure Chemical Industries, Ltd.), then injected into a cell counting board (Funakoshi Co., Ltd.), and observed under a phase-contrast microscope (OLYMPUS Co., Ltd.) to count the cell density. Cell viability was assessed by classifying cells that were not stained due to their ability to exclude trypan blue dye as live cells and cells that stained blue with the dye as dead cells. Cell viability was calculated using the formula: live cell density ÷ total cell (live cell + dead cell) density. The results are shown in Tables 8 and 9.

[0189]

[0190]

[0191] The results shown in Tables 8 and 9 indicate that cell culture bag 1 has a higher cell culture efficiency than cell culture bag 2.

[0192] 3-2-2. Test 2 3 x 10 ６Human peripheral blood mononuclear cells (PBMC) (manufactured by LONZA) were suspended in 6 ml of RPMI 1640 medium (manufactured by Gibco) containing 10% FCS, 1% penicillin / streptomycin, 50 ng / ml CD3 monoclonal antibody (manufactured by Thermo Fisher Scientific), 2 μg / ml CD28 monoclonal antibody (manufactured by Thermo Fisher Scientific), and 500 IU / ml recombinant human interleukin 2 (manufactured by Proteintech), and then introduced into the sterilized cell culture bag 3 with ports and cell culture bag 4. The cell culture bags containing the cell suspension were incubated at 37°C and 5% CO ２ CO maintained at ２ The cells were placed in an incubator and cultured for 20 days. After 3, 6, 9, 12, 15, and 17 days, the entire cell suspension was collected from the port using a syringe. The collected cell suspension was centrifuged at 400 g for 10 minutes, and 3 ml of the supernatant was removed. 3 ml of RPMI 1640 medium containing 10% FCS, 1% penicillin / streptomycin, and 500 IU / ml recombinant human interleukin-2 was added, suspended, and reintroduced into the cell culture bag.

[0193] Twenty days after the start of culture, cell density and cell viability were calculated for the cell suspension using the trypan blue exclusion method. Specifically, the cell suspension was mixed with 0.4 w / v% trypan blue solution (Gibco), then injected into a cell counting board (Wakembee Tech Co., Ltd.), and observed using a phase-contrast microscope (OLYMPUS) to count the cell density. Cell viability was assessed by classifying cells that were not stained due to their ability to exclude trypan blue dye as live cells and cells that stained blue with the dye as dead cells. Cell viability was calculated using the formula: live cell density ÷ total cell (live cell + dead cell) density. The results are shown in Tables 10 and 11.

[0194]

[0195]

[0196] The results shown in Tables 10 and 11 show that cell culture bag 3 has a higher cell culture efficiency than cell bag 4.

[0197] This application claims priority to Japanese Patent Application No. 2023-170083, filed September 29, 2023, and Japanese Patent Application No. 2023-214004, filed December 19, 2023. The matters described in the specification, claims, and drawings of those applications as originally filed are incorporated herein by reference.

[0198] The cell container according to the present invention can be applied to the culture of various cells.

[0199] REFERENCE SIGNS LIST 100 Cell container 110 Bag member 112, 114 Resin film 116 Joint portion 200 Port member 210 Main body portion 212 Narrow cylindrical portion 214 Wide cylindrical portion 220 Passage 300 Thermal adhesive film 310 Elastomer layer 320 Thermoplastic resin layer 330 Elastomer layer

Claims

1. A cell container having a resin bag member and a port member including a molded body of a resin composition that connects the inside and outside of the bag member, wherein the bag member includes a multi-layer resin film including, from the outside to the inside of the cell container, a surface layer, a core layer and a heat seal layer in that order, and the heat seal layer includes a 4-methyl-1-pentene-based copolymer.

2. The cell container according to claim 1, wherein the core layer comprises a 4-methyl-1-pentene copolymer or a styrene elastomer.

3. The cell container described in claim 1, wherein the 4-methyl-1-pentene copolymer contained in the heat seal layer has 80 mol % or more and 99 mol % or less of structural units derived from 4-methyl-1-pentene, and 1 mol % or more and 20 mol % or less of structural units derived from an α-olefin having from 2 to 20 carbon atoms, excluding structural units derived from 4-methyl-1-pentene, relative to all structural units.

4. The bag member has an oxygen permeability of 3.0 L / (m at 23°C. ２ ・day・atm) or more 100.0L / (m ２ The cell container according to claim 1 , which is formed from the resin film having a thermal conductivity of 100 / single-day atm or less.

5. The bag member has a sterility assurance level (SAL) of 10 for medical devices measured in accordance with BS EN556-1:2001. －３ The cell container of claim 1 , wherein:

6. The cell container according to claim 1, wherein the resin composition molded body contained in the port member is a resin composition molded body containing at least one resin selected from the group consisting of 4-methyl-1-pentene copolymers, polyethylene, and polypropylene.

7. The cell container according to claim 1, wherein the resin composition molded article contained in the port member is a resin composition molded article having a peel strength of 20 N / 15 mm or more and 100 N / 15 mm or less as measured by a 180° peel test conducted under the following measurement conditions: Measurement conditions: A 65 mm x 65 mm x 1.0 mm press sheet obtained from the resin composition is heat-sealed to a flat surface of a 65 mm x 65 mm x 0.3 mm substrate formed from a 4-methyl-1-pentene copolymer consisting of 4-methyl-1-pentene structural units and propylene structural units at a temperature of 240°C, a pressure of 0.1 MPa, a press time of 5 seconds, and a seal width of 10 mm, and then a 180° peel test is conducted at 23°C using a tensile tester.

8. The cell container according to claim 1, wherein the resin composition molded body contained in the port member is a resin composition molded body having a total light transmittance of 85% or more when measured on a press sheet having a thickness of 1 mm.

9. The cell container according to claim 1, wherein the resin composition molded body contained in the port member is a resin composition molded body having a durometer hardness type D of 30 or more and 80 or less when measured on a press sheet having a thickness of 1 mm, as measured in accordance with ASTM D2240.

10. The cell container according to claim 1, wherein the port member has a thin cylindrical portion having a shorter outer circumferential length and a thick cylindrical portion having a longer outer circumferential length, and at least the thick cylindrical portion includes a molded body of the resin composition.

11. The cell container according to claim 1, wherein the port member is joined to the bag member via a single-layer or multi-layer thermal adhesive film having an elastomer layer containing at least a styrene-based elastomer.

12. The cell container according to claim 11, wherein the thermal adhesive film is arranged to circumferentially surround the port member, which is cylindrical, and the elastomer layer is arranged to surround the entire periphery of the port member.

13. The cell container according to claim 12, wherein the thermal adhesive film is welded to the entire periphery of the port member.

14. The cell container of claim 11, wherein the thermal adhesive film has a layer comprising a 4-methyl-1-pentene copolymer.

15. The cell container of claim 14, wherein the layer containing the 4-methyl-1-pentene copolymer is a sealant layer.

16. The cell container according to claim 11, wherein the bonding width of the thermal adhesive film to the port member in the direction in which the port member communicates between the inside and outside of the bag member is 1 mm or more and 50 mm or less.

17. A method for manufacturing a cell container, comprising: a step of arranging resin bag members containing at least a 4-methyl-1-pentene copolymer with their ends overlapping; and a step of heat-sealing the overlapped bag members to form a bag, wherein in the overlapping and arranging step, a port member is arranged between the overlapped ends; and in the processing step, the bag members with the port member arranged therebetween are heated and pressurized to join the bag member and the port member.

18. A method for manufacturing a cell container as described in claim 17, wherein in the overlapping and arranging step, a single-layer or multi-layer thermal adhesive film having an elastomer layer containing at least a styrene-based elastomer is arranged between the port member and the bag member, and in the processing step into a bag shape, the bag member and the port member are joined via the thermal adhesive film.

19. A port member comprising a resin bag member containing at least a 4-methyl-1-pentene copolymer and a port member welded to the resin bag member to communicate between the inside and outside of the bag member, the port member comprising a molded article of a resin composition containing a 4-methyl-1-pentene copolymer, the resin composition having a total light transmittance of 85% or more when measured on a press sheet having a thickness of 1 mm.

20. A method for culturing cells, comprising the steps of: introducing cells into a cell container according to any one of claims 1 to 16; and culturing the cells in the cell container.

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