JP5919776B2 - Method for producing cell culture vessel - Google Patents

Description

  The present invention relates to a method for producing a cell culture container on which a functional substrate having a functional organic compound layer is fixed.

  Patent Document 1 discloses a cell culture container such as a petri dish having a surface coated with a functional organic compound layer such as a temperature-responsive polymer. By using this cell culture container, it is possible to easily peel off and recover from the cell support without destroying the cultured / proliferated cells simply by changing the temperature. However, as described in Patent Document 1, it is necessary to perform surface treatment separately by batch treatment on a cell culture container such as a petri dish to provide a functional compound layer from the viewpoint of workability. There is still room for improvement.

  Therefore, as in Patent Document 2, etc., a functional substrate having a functional compound layer on the surface and having a shape such as a film shape or a plate shape is separately prepared, and the functional substrate is placed on the bottom surface of the container with an adhesive or There has been proposed a technique for forming a target cell culture container by fixing it with an adhesive or the like. According to this technique, workability is enhanced as compared with the case where the functional organic compound layer is provided by batch processing of the cell culture containers individually.

  As a means for joining the functional substrate to the cell culture vessel, there is a means for joining them together with an adhesive. For this purpose, a functional substrate comprising at least a base material layer, a functional compound layer disposed on one side of the base material layer, and an adhesive layer disposed on the other side of the base material layer, It is common to join to a cell culture container through the adhesive layer. This pressure-sensitive adhesive layer is an adhesive or a layer containing a pressure-sensitive adhesive, and is a layer made of a substance having a self-adhesive property such as a polyacrylic acid ester-based adhesive as an adhesive and an acrylic pressure-sensitive adhesive as a pressure-sensitive adhesive Usually it is.

JP-A-2-21865 JP 2010-98799 A

  When using a functional substrate with a pressure-sensitive adhesive layer that always shows adhesiveness, it is usual to protect the pressure-sensitive adhesive layer with a release film until application, and then apply it to the container after removing the release film. . A film-like functional substrate (functional film) is preferable because it can be mass-produced by a roll-to-roll method, but a functional film having a pressure-sensitive adhesive layer that always shows adhesiveness is used as a release film. It is virtually impossible to produce in a roll-to-roll manner without protection.

  For this reason, the method of sticking a functional substrate having a pressure-sensitive adhesive layer that always exhibits adhesiveness to a cell culture container requires a step for removing the release film, the release film is generated as waste, There are problems such as.

  Then, this invention aims at providing the efficient method for joining a functional base | substrate to a container main body and manufacturing a cell culture container which does not require the protection by a peeling film.

The present invention requires protection by a release film by using a bonding layer that does not substantially exhibit adhesiveness in a dry state and exhibits adhesiveness in a wet state with a solvent, instead of an adhesive layer that always exhibits adhesiveness. An efficient method for manufacturing a cell culture container by bonding a functional substrate to a container body is provided. Specifically, the following invention is included.
(1) A method for producing a cell culture container comprising a container part for containing cells and a medium,
The container part
A container body member;
A functional substrate comprising at least a base material layer and a functional organic compound layer disposed on one side of the base material layer;
The functional substrate is a bonding layer that exists between the base material layer and the surface of the container body member so that the functional organic compound layer faces the space for accommodating cells and culture medium on the surface of the container body member. Are joined via
The bonding layer is a layer that does not substantially exhibit adhesiveness in a dry state and exhibits adhesiveness in a wet state with a solvent,
The following steps:
A functional substrate in which the bonding layer is further disposed on the other side of the base material layer, with the bonding layer wetted with a solvent, the bonding layer and the surface are disposed on the surface of the container body member. A contact step of bringing the functional base into contact with the surface of the container body member through the bonding layer by volatilizing the solvent while maintaining the contact state. Method.
(2) The contact process is
A step of preparing a functional substrate in a state where the bonding layer is wetted with a solvent and a container body member,
Contacting the functional substrate with the surface of the container body member.
(3) The contact process is
A step of preparing a functional substrate in a state where the bonding layer is dried and a container body member in a state where a solvent is present on the surface;
Contacting the functional substrate with the surface of the container body member.
(4) A method for producing a cell culture container comprising a container part for containing cells and a medium,
The container part
A container body member;
A functional substrate comprising at least a base material layer and a functional organic compound layer disposed on one side of the base material layer;
The functional substrate is a bonding layer that exists between the base material layer and the surface of the container body member so that the functional organic compound layer faces the space for accommodating cells and culture medium on the surface of the container body member. Are joined via
The bonding layer is a layer that is weakly tacky or does not substantially exhibit tackiness in a dry state, and exhibits tackiness in a wet state with a solvent,
The following steps:
A contact step in which the functional substrate is brought into contact with the container body member having the bonding layer disposed on the surface so that the base layer and the bonding layer are in contact with each other while the bonding layer is wetted with a solvent. As well as
A method comprising a bonding step of bonding the functional substrate to the surface of the container body member via the bonding layer by volatilizing the solvent while maintaining a contact state.
(5) The functional substrate is a film-like functional substrate,
The long functional substrate wound in a roll shape with the bonding layer dried in advance is fed out, and the fed out functional substrate is cut into a shape to be bonded to the container body member. The method according to any one of (1) to (3), further comprising a cutting step.
(6) The functional substrate is a film-like functional substrate,
The method further includes a cutting step in which the long functional substrate wound in a roll is prepared, and the functional substrate is cut into a shape to be joined to the container body member. ) Method.

  ADVANTAGE OF THE INVENTION According to this invention, the efficient method for manufacturing a cell culture container by joining a functional base | substrate to a container main body which does not require protection by a peeling film is provided.

  In addition, the pressure-sensitive adhesive layer that always exhibits adhesiveness has a problem that air bubbles tend to remain on the application surface of the functional substrate and the container body member because of high adhesiveness at the time of application, The bonding layer that has been wetted with a solvent and developed a viscosity has a relatively low adhesiveness at the time of application, and therefore, it is difficult for bubbles to remain on the application surface.

It is a perspective view which shows one Embodiment of the cell culture container manufactured by this invention. It is I-I 'sectional drawing of FIG. 1A. It is II-II 'sectional drawing of FIG. 1A. It is a perspective view for demonstrating the manufacturing process of the cell culture container shown to FIG. 1A-FIG. 1C. It is a perspective view for demonstrating the manufacture process of another embodiment of the cell culture container manufactured by this invention. It is a perspective view which shows another embodiment of the cell culture container manufactured by this invention. FIG. 4B is a cross-sectional view taken along the line III-III ′ of FIG. 4A. It is sectional drawing of the functional base | substrate used for 2nd Embodiment of the method of this invention. It is sectional drawing of the functional base | substrate used for 1st Embodiment of the method of this invention. It is sectional drawing for demonstrating an example of 1st Embodiment of the manufacturing method of the cell culture container of this invention. It is sectional drawing for demonstrating another example of 1st Embodiment of the manufacturing method of the cell culture container of this invention. FIG. 3 is a schematic top view of a long functional substrate for producing a film-like functional substrate that can be used in the first embodiment of the method for producing a cell culture container of the present invention. It is IV-IV 'sectional drawing of FIG. 9A. It is sectional drawing for demonstrating an example of 2nd Embodiment of the manufacturing method of the cell culture container of this invention. FIG. 5 is a schematic top view of a long functional substrate for producing a film-like functional substrate that can be used in the second embodiment of the method for producing a cell culture container of the present invention. FIG. 11B is a cross-sectional view taken along the line V-V ′ of FIG. 11A.

  Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. Each figure is a schematic diagram, and the size and shape of each part are appropriately exaggerated for easy understanding.

<Shape of cell culture vessel>
First, the overall shape of the cell culture container produced in the present invention will be described.
The cell culture container produced by the present invention includes at least a container part for containing cells and a medium, and further includes a lid or the like as appropriate.

  As shown in FIGS. 1A to 1C, the container portion may have a shape in which a space for containing cells and a medium is closed by a wall surface, or one end of the space as shown in FIGS. 4A and 4B. The shape may be open.

  A preferred embodiment of a cell culture vessel is shown in FIGS. 1A-1C. A container unit 100 shown in FIG. 1A includes a container body member 103 composed of a bottom part 101 and a side wall part 102 erected on the periphery of the bottom part 101, and a bottom part 101 joined to an upper end part of the container body member 103. And a top surface member 104 disposed to face each other. A through-hole 105 is formed in a part of the side wall portion 102, and the container portion has a shape called “flask type” including a neck portion 106 that extends from the periphery of the through-hole 105 to the outside of the container portion. A locking part 107 for locking the lid 110 is formed on the neck part 106 of the container part 100, and the lid 110 is detachably mounted via the locking part 107. A flask type cell culture container 120 is formed by combining the container part 100 and the lid 110.

  FIG. 1B shows a cross-sectional view taken along the line I-I ′ of the container 100, and FIG. 1C shows a cross-sectional view taken along the line II-II ′. The container main body member 103 composed of the bottom 101 and the side wall 102 of the container 100 includes an opening 108 that is defined by the bottom 101 and the side wall 102 and opened upward. The opening 108 is closed by joining the top surface member 104, and as a result, a space 130 for accommodating cells and a medium is formed. A functional substrate 140 is fixed to the bottom surface of the container main body member 103 on the opening 108 side (that is, on the bottom 101 facing the space 130).

  In order to form the container part 100, first, as shown in FIG. 2, the functional base 140 is joined to the inner bottom surface of the container body member 103 composed of the bottom part and the side wall part, and then the container body member 103 is joined to the container body member 103. The top member 104 is joined.

  FIG. 3 shows a manufacturing process of another embodiment of a flask-type container similar to FIGS. 1A to 1C. In this embodiment, the container main body member 109 is formed in a flat plate shape, and the functional substrate 140 is fixed on the bottom surface (surface) of the flat plate main body member 109. And the side wall member 111 and the top | upper surface member 104 are joined with respect to the container main body member 109 to which the functional base | substrate 140 was fixed, and the container part which has the same outer shape as FIG. 1A-FIG. 1C is obtained. In FIG. 3, it goes without saying that either the joining process of the container body member 109 and the side wall member 111 and the joining process of the side wall member 111 and the top surface member 104 may be performed first.

  As another embodiment of the container part, as shown in FIGS. 4A and 4B, a dish-shaped container part including a container body member 203 composed of a bottom part 201 and a side wall part 202 standing on the periphery of the bottom part 201. 200. The container body member 203 includes an opening 204 that is defined by the bottom 201 and the side wall 202 and is opened upward, and the opening 204 functions as a space 220 for containing cells and a culture medium. A functional base 210 is fixed to the bottom surface of the container main body member 203 on the opening 204 side (that is, on the bottom 201 facing the space 220).

  In the present invention, the “container body member” is a member corresponding to the whole or a part of the container body that provides a surface to which the functional substrate is bonded. As shown in FIGS. 2 and 3, in the embodiment in which the container main body member is a member corresponding to a part of the container main body, the remaining portion constituting the container main body can be appropriately joined to the container main body member.

<Material of cell culture container>
The material forming the members of the cell culture container such as the container main body member, the side wall member, the top surface member, the neck and the lid is not particularly limited, and materials generally used in cell culture can be used. For example, polystyrene resin, polyester resin, polyethylene resin, polyethylene terephthalate resin, polypropylene resin, ABS resin, nylon, acrylic resin, fluorine resin, polycarbonate resin, polyurethane resin, methylpentene resin, phenol resin, melamine resin, epoxy resin, vinyl chloride A resin material such as a resin, a resin material containing at least one of the above-described surface hydrophilized treatment, and an inorganic material such as glass or quartz are preferable, but a resin material is preferable. The resin material is preferably a polystyrene resin or a polyethylene terephthalate resin.

  The container body member is preferably made of a transparent material through which the cultured cells can be observed from outside the container.

<Functional substrate>
As shown in FIG. 5, the cross section along the thickness direction of the functional substrates 140 and 210 in the present invention includes at least a base material layer 502 and a functional organic compound layer 501 disposed on the base material layer 502. Prepare. The functional substrate is typically a transparent functional substrate. Here, the “base” may be a film or a plate-like body as long as it has a predetermined structure. The film-like functional substrate (hereinafter sometimes referred to as “functional film”) is preferably flexible so that it can be wound into a roll. A flexible functional film is preferable because mass production by a roll-to-roll method is easy.

  The functional substrate can be manufactured by forming the functional organic compound layer 501 on the base material layer 502 by an appropriate method. Between the base material layer 502 and the functional organic compound layer 501, one or more other layers (for example, a primer layer described later) may be present as necessary.

  The shape of the functional substrate can be any shape depending on the shape of the region of the member to be fixed. For example, it may be a polygon such as a triangle, a rectangle (rectangle, square, parallelogram, rhombus, etc.), a pentagon, a hexagon, a heptagon, an octagon, a circle, an ellipse, or the like.

<Base material layer>
The base material layer 502 is appropriately selected according to the final functional substrate. If the functional substrate is plate-like, a plate-like substrate layer is used, and if the functional substrate is film-like, a film-like substrate layer (hereinafter referred to as “film substrate layer”) is used.

  The base material layer 502 only needs to include a material capable of forming the above-described functional organic compound layer 501 on one surface, and the type of the material is not particularly limited. Typically, as a material for the base layer, polyethylene terephthalate (PET), polystyrene (PS), polycarbonate (PC), TAC (triacetylcellulose), polyimide (PI), nylon (Ny), low density polyethylene (LDPE) ), Medium density polyethylene (MDPE), polyvinyl chloride, polyvinylidene chloride, polyphenylene sulfide, polyether sulfone, polyethylene naphthalate, polypropylene, acrylic resin and other inorganic materials such as glass and quartz, A resin material is preferred.

  The surface of the base material layer 502 on which the functional organic compound layer 501 is formed can be a surface that has been subjected to easy adhesion treatment. “Easy adhesion treatment” refers to treatment with an easy adhesive such as polyester, acrylic ester, polyurethane, polyethyleneimine, silane coupling agent, perfluorooctane sulfonic acid (PFOS), and the like.

  The thickness of the base material layer 502 can be selected as appropriate. When the base material layer is a film base material layer, its thickness (when the film base material layer includes an easy adhesion layer in addition to the base material layer, the total thickness of the film base material layer including the easy adhesion layer) Is not particularly limited, but is preferably a thickness imparting flexibility, for example, 5 to 500 μm, more preferably 20 to 500 μm, and particularly preferably 50 to 250 μm.

<Functional organic compound layer>
The organic compound constituting the functional organic compound layer 501 is not particularly limited as long as it has a desired function, but more preferably, the organic compound can be changed from cell adhesiveness to cell nonadhesiveness by a predetermined stimulus. Examples include stimuli-responsive polymers having a possible surface, and hydrophilic compounds such as ethylene glycol chains composed of one or more ethylene glycol units (CH ₂ —CH ₂ —O).

  The film thickness of the functional organic compound layer 501 may be, for example, in the range of 0.5 nm to 300 nm, and particularly preferably in the range of 1 nm to 100 nm.

  Hereinafter, preferred embodiments of the “stimulus responsive polymer layer” and the “hydrophilic compound layer” will be described.

<Stimulus responsive polymer layer>
The functional organic compound layer 501 is particularly preferably a stimulus-responsive polymer layer. The stimulus-responsive polymer layer is a layer containing a polymer in which the degree of surface cell adhesion is changed by a predetermined stimulus. Examples of the stimulus responsive polymer include a temperature responsive polymer, a pH responsive polymer, an ion responsive polymer, and a photoresponsive polymer. Among these, a temperature-responsive polymer is preferable because it is easy to give a stimulus.

  As the temperature-responsive polymer, for example, a polymer that exhibits cell adhesion at a temperature at which cells are cultured and exhibits cell non-adhesion at a temperature at which the produced cell sheet is peeled may be used. For example, a temperature-responsive polymer has improved affinity to surrounding water at temperatures below the critical dissolution temperature, and the polymer takes up water and swells to make it difficult for cells to adhere to the surface (cell non-adhesiveness). It is preferable to use a material exhibiting a property (cell adhesiveness) that makes the polymer shrink and easily adheres cells to the surface when water is desorbed from the polymer at a temperature equal to or higher than the same temperature. Such a critical solution temperature is called a lower critical solution temperature. A temperature-responsive polymer having a lower critical solution temperature T of 0 ° C. to 80 ° C., more preferably 0 ° C. to 50 ° C. may be used. This is because the cells can be stably cultured when T is 0 ° C to 80 ° C.

  Suitable temperature-responsive polymers include acrylic polymers or methacrylic polymers. Specific examples of suitable temperature-responsive polymers include poly-N-isopropylacrylamide (T = 32 ° C.), poly-Nn-propyl acrylamide (T = 21 ° C.), and poly-Nn-propyl methacrylamide. (T = 32 ° C.), poly-N-ethoxyethyl acrylamide (T = about 35 ° C.), poly-N-tetrahydrofurfuryl acrylamide (T = about 28 ° C.), poly-N-tetrahydrofurfuryl methacrylamide (T = About 35 ° C.), and poly-N, N-diethylacrylamide (T = 32 ° C.).

  As a monomer for forming these polymers, a monomer that can be polymerized by irradiation with radiation can be used. Examples of the monomer include (meth) acrylamide compounds, N- (or N, N-di) alkyl-substituted (meth) acrylamide derivatives, (meth) acrylamide derivatives having a cyclic group, and vinyl ether derivatives. More than seeds may be used. When a single monomer is used alone, the polymer formed on the substrate is a homopolymer, and when multiple monomers are used together, the polymer formed on the substrate is a heteropolymer. Both forms are encompassed by the present invention.

  In addition, when it is necessary to adjust T depending on the type of proliferating cell, when it is necessary to enhance the interaction between the coating substance and the cell culture support, and the balance between the hydrophilicity and hydrophobicity of the cell support is adjusted. If necessary, other monomers other than those described above may be further added for copolymerization. Further, a graft or block copolymer of the above polymer used in the present invention and another polymer, or a mixture of the polymer of the present invention and another polymer may be used. Moreover, it is also possible to crosslink within a range where the original properties of the polymer are not impaired.

  As the pH responsive polymer and the ion responsive polymer, those suitable for the cell sheet to be prepared can be appropriately selected.

  The stimulus-responsive polymer layer is prepared by preparing a coating composition containing a monomer that is polymerized to form a target stimulus-responsive polymer and an organic solvent that can dissolve the monomer. A coating film is formed by coating on the surface of the layer, and then the monomer in the coating film is polymerized by an appropriate means such as radiation irradiation to form a polymer. And a grafting reaction between them.

<Hydrophilic compound layer>
As another embodiment of the functional organic compound layer, hydrophilicity such as an ethylene glycol chain composed of one or more ethylene glycol units (an ethylene glycol chain composed of a plurality of ethylene glycol units can be referred to as a “polyethylene glycol chain”). A layer of a functional compound. The terminal of the ethylene glycol chain may be in a form blocked with a hydroxyl group, or the terminal of the ethylene glycol chain may be in a form in which another substance such as a biological substance is linked by a covalent bond. A layer containing an ethylene glycol chain whose end is blocked with a hydroxyl group can provide a hydrophilic surface to which cells are difficult to adhere.

  Examples of biological substances that can be covalently bonded to the end of an ethylene glycol chain include antigens, antibodies, DNA, RNA, peptides, hormones, enzymes, cytokines, sugar chains, lipids, coenzymes, enzyme inhibitors, cells, and other functions. The protein which has is included. Furthermore, the low molecular weight compound which has affinity with such a biological substance, and a high molecular compound are also contained in the range of a biological substance.

  In order to immobilize a layer of a hydrophilic compound such as an ethylene glycol chain on the surface of a resin base material layer, it can be physically adsorbed on the surface of the resin base material layer in advance. Then, a primer layer containing polysiloxane containing a functional group capable of forming a covalent bond by reacting with the terminal hydroxyl group of the ethylene glycol chain is provided. The functional group on the side chain of the polysiloxane is preferably a glycidyl group or an epoxy group. The primer layer can be formed by applying a silanol compound having a desired side chain to the surface of the base material layer, and performing condensation polymerization on the surface to convert it into polysiloxane.

  Next, a functional group of the primer layer is reacted with a hydroxyl group of polyethylene glycol in which two or more ethylene glycol or ethylene glycol units are repeated to form a covalent bond, thereby immobilizing the ethylene glycol chain. At this time, ethylene glycol or polyethylene glycol containing a catalytic amount of concentrated sulfuric acid is brought into contact with the primer layer. A layer containing ethylene glycol chains whose ends are blocked with hydroxyl groups is formed in this way.

  Furthermore, if necessary, at least one functional group capable of forming a covalent bond with another substance is directly or indirectly linked to one end of the ethylene glycol chain. The method for introducing the functional group is not particularly limited.

<Junction layer>
One of the features of the present invention is that the base layer 502 of the functional substrate and the surfaces of the container main body members 103, 109, and 203 are bonded via the bonding layer 503.

  In the first embodiment of the present invention, the bonding layer 503 is disposed in advance on the functional substrate. That is, the functional substrates 140-1 and 210-1 used in the first embodiment of the present invention include the functional organic compound layer 501 of the base material layer 502 in addition to the base material layer 502 and the functional organic compound layer 501. Is prepared in advance in a state in which a bonding layer 503 is further provided on the side where no is formed (FIG. 6).

  In the second embodiment of the present invention, the bonding layer 503 is disposed in advance on the container body member. Therefore, the functional bases 140 and 210 used in the second embodiment of the present invention need only have two layers shown in FIG.

  The bonding layer 503 is a layer that does not substantially exhibit adhesiveness in a dry state and exhibits adhesiveness in a wet state with a solvent. Since such a bonding layer does not have adhesiveness in a dry state, it is not necessary to protect the surface with a release film. A functional substrate or container body member provided with such a bonding layer is easy to handle and suitable for mass production. In particular, since the film-like and long functional substrate provided with the bonding layer can be wound into a roll and fed out from the roll without the release film, a roll-to-roll described later Continuous treatment by the method is easy and preferable.

  As a material for forming the bonding layer 503, the adhesive layer exhibits adhesiveness in a wet state in which water, isopropyl alcohol, propanol, ethanol, methanol, or the like, preferably water or isopropyl alcohol is absorbed, and substantially in the dry state. Examples thereof include materials not shown, and specific examples include polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylamide, and acrylic acid copolymer. In addition to these materials, the bonding layer may appropriately contain other components such as a crosslinking agent and a filler.

  In the second embodiment of the present invention, the bonding layer 503 provided on the surface of the container body member is not required until it has no adhesiveness in the dry state, and adheres to the extent that protection by the release film is not necessary in the dry state. The same effect can be obtained even if the layer is weak. Examples of such a bonding layer 503 include a material that exhibits adhesiveness in a wet state in which the solvent is absorbed, and that has low adhesiveness to the extent that protection by a release film is not necessary in a dry state. Specifically, polyvinyl alcohol, The thing etc. which added the tackifier to any one of polyvinylpyrrolidone, polyacrylamide, and an acrylic acid copolymer are mentioned.

  The thickness of the bonding layer 503 can be determined as appropriate so that the functional substrate and the container body member can be bonded, and can generally have a thickness of 1 to 20 μm in a dry state. .

  A method for forming the bonding layer 503 on the surface of the base layer of the functional substrate or the container body member is not particularly limited, and for example, a printing method or a spin coating method can be used. When the bonding layer 503 is formed of a water-soluble polymer material, the base layer or the container body member of the functional substrate is used for the purpose of preventing the bonding layer from eluting into the cell culture medium during cell culture. It is also possible to form a bonding layer 503 by covalently bonding a polymer material to the surface by graft polymerization.

  The bonding layer 503 ′ (see FIG. 7 and the like) wetted with a solvent has a lower viscosity than a pressure-sensitive adhesive layer that is always sticky. The pressure-sensitive adhesive layer that always exhibits adhesiveness has a problem that bubbles are likely to remain on the application surface due to its high adhesiveness at the time of application. The use of the bonded layer 503 ′ is also advantageous for reducing the remaining bubbles.

<Manufacturing method>
Hereinafter, each of 1st Embodiment and 2nd Embodiment of the manufacturing method of the cell culture container of this invention is demonstrated.

<First Embodiment>
In the first embodiment, a base material layer 502, a functional organic compound layer 501 disposed on one side of the base material layer 502, and a bonding layer disposed on the other side of the base material layer 502 shown in FIG. In this embodiment, the functional substrates 140-1 and 210-1 having 503 are used.

The first embodiment of the method for producing a cell culture container of the present invention comprises:
A contact step in which the functional substrate having the structure is brought into contact with the surface of the container body member so that the bonding layer and the surface are in contact with each other in a state where the bonding layer 503 is wetted with a solvent, and while maintaining the contact state It includes at least a bonding step of bonding the functional substrate to the surface of the container main body member via the bonding layer by volatilizing the solvent.

  7 and 8, the functional base 140-1 is joined to the container main body member 103 and then the top member 104 is joined by the method according to the first embodiment, and then the flask type shown in FIGS. 1A to 1C. A specific example of manufacturing the cell culture container (the direction of II ′ cross-sectional view in FIG. 1A) will be described.

  In a typical example of the first embodiment, as shown in FIG. 7, a predetermined solvent is applied to the bonding layer 503 in a dry state of the functional base 140-1 before being bonded to the container main body member 103 and is wetted. (FIG. 7B). The wet bonding layer 503 'exhibits adhesiveness as described above. Next, the functional substrate 140-1 and the container body member 103 are brought into contact with each other so that the wet bonding layer 503 'is in contact with a predetermined surface of the separately prepared container body member 103 (FIG. 7C). Next, the solvent is volatilized while maintaining the contact state, and the base material layer 502 is bonded to the bottom surface of the container body member 103 via the bonding layer 503 (FIG. 7D). Next, the top plate member 104 is joined to the upper end edge of the container body member 103 to complete the flask-type container unit 100 (FIG. 7E). The conditions of the joining step for volatilizing the solvent are not particularly limited as long as the solvent can be volatilized, but preferred examples include natural drying in a low-humidity environment, air drying, and use of a dry hood.

  Typically, the solvent is not completely removed by volatilization in the bonding process of the first embodiment and the second embodiment, and FIGS. 7D, 7E, 8C, 8D, and 10D after the bonding process are performed. An appropriate amount of solvent remains in the bonding layer 503 in FIG. 10E. The degree of volatilization of the solvent in the first embodiment and the second embodiment of the present invention can be adjusted as appropriate so that the functional substrate after the volatilization of the solvent and the container body member are joined with sufficient strength.

  In another example of the first embodiment, as shown in FIG. 8, a solvent 810 is disposed on a predetermined surface of the container body member 103 on which the functional substrate is provided by coating, dropping, or the like, and prepared separately here. The functional substrate 140-1 having the dried bonding layer 503 is brought into contact so that the bonding layer 503 is wetted by the solvent 810 to form a wet bonding layer 503 ′ (FIG. 8B). Next, the solvent is volatilized while maintaining the contact state, and the base material layer 502 is bonded to the bottom surface of the container body member 103 via the bonding layer 503 (FIG. 8C). Next, the top plate member 104 is joined to the upper edge of the container body member 103 to complete the flask-type container unit 100 (FIG. 8D). The amount of the solvent 810 disposed on the surface of the container body member 103 is not particularly limited as long as it is a sufficient amount to wet the bonding layer 503 and develop adhesiveness. As the solvent 810, a solvent that can wet the bonding layer 503 and develop adhesiveness as illustrated above can be appropriately selected and used depending on the material of the bonding layer 503.

  The functional substrate 140-1 for use in the first embodiment shown in FIGS. 7 and 8 is preferably in the form of a film. The film-like functional substrate 140-1 can be manufactured from a long film-like functional substrate 900 including a functional organic compound layer 501, a base material layer 502, and a bonding layer 503, as shown in FIG. it can. The long film-like functional substrate 900 can be wound into a roll with the bonding layer 503 dried, and does not require protection with a release film. A long film-like functional substrate 900 wound in a roll shape is appropriately fed out, and the fed-out functional substrate 900 is cut into a piece of a predetermined shape to be joined to the container body member 103 to obtain a sheet-like shape. The functional substrate 140-1 can be obtained (cut process). It is also possible to perform a roll-to-roll process in which the remaining portion of the long film-like functional substrate 900 after the functional substrate 140-1 is cut off is rolled again. The cutting step may be a step of cutting the sheet-like functional substrate 140-1 from the long film-like functional substrate 900 by cutting, or as the long film-like functional substrate 900, the functional substrate. The cutting process may be a process of removing the functional substrate 140-1 along these cuts, using perforations, half-cut lines and the like so that 140-1 can be cut off. . As shown in FIG. 7, when the bonding layer 503 on the functional substrate 140-1 is wetted, the bonding layer 503 of the long functional substrate 900 fed out from the roll is formed before the cutting step. A sheet-like functional substrate 140-1 having a wet bonding layer 503 ′ can be manufactured by applying a solvent and moistening, and then executing a cutting process. Alternatively, a wet bonding layer 503 ′ can be formed by applying a solvent to the sheet-like functional substrate 140-1 after the cutting process.

Second Embodiment
In the second embodiment, a functional substrate having a base material layer 502 and a functional organic compound layer 501 disposed on one side of the base material layer 502 shown in FIG. 5 and a bonding layer on the surface are provided. It is embodiment using a container main body member.

The second embodiment of the method for producing a cell culture container of the present invention comprises:
A contact step in which the functional substrate is brought into contact with the container body member on which the bonding layer 503 is disposed in a state where the bonding layer is wetted with a solvent so that the base material layer and the bonding layer are in contact with each other; And
It includes at least a bonding step of bonding the functional substrate to the surface of the container body member via the bonding layer by volatilizing the solvent while maintaining the contact state.

  Specifically, as shown in FIG. 10, a container body member 103 having a bonding layer 503 disposed on the bottom surface is prepared, and the bonding layer 503 is wetted with a solvent to form a bonding layer 503 ′ (FIG. 10B). . The wet bonding layer 503 'exhibits adhesiveness as described above. A functional substrate 140 having a separately prepared base material layer 502 and a functional organic compound layer 501 is brought into contact with the container body member 103 so that the base material layer 502 is in contact with the wet bonding layer 503 ′. (FIG. 10C). Next, the solvent is volatilized while maintaining the contact state, and the base material layer 502 is bonded to the bottom surface of the container body member 103 via the bonding layer 503 (FIG. 10D). Next, the top plate member 104 is joined to the upper end edge of the container body member 103 to complete the flask-type container unit 100 (FIG. 10E).

  The functional substrate 140 for use in the second embodiment shown in FIG. 10 is preferably in the form of a film. The film-like functional substrate 140 can be manufactured from a long film-like functional substrate 1100 including a functional organic compound layer 501 and a base material layer 502 as shown in FIG. The long film-like functional substrate 1100 can be wound into a roll and does not need to be protected by a release film. A long film-like functional substrate 1100 wound in a roll shape is appropriately fed out, and the fed-out functional substrate 1100 is cut into sheet pieces of a predetermined shape to be joined to the container body member 103 to obtain a sheet-like shape. The functional substrate 140 can be obtained (cut process). It is also possible to perform a roll-to-roll process in which the remaining portion of the long film-like functional substrate 1100 after the functional substrate 140 is cut off is wound into a roll. The cutting step may be a step of cutting the sheet-like functional substrate 140 from the long film-like functional substrate 1100 by cutting, or the functional substrate 140 is used as the long film-like functional substrate 1100. A cut provided in advance such as a perforation or a half cut line so as to be cut off is used, and the cutting step may be a step of removing the functional substrate 140 along these cuts.

100, 200 Container part 101, 201 Bottom part 102, 202 Side wall part 103, 109, 203 Container body member 140, 140-1, 210, 210-1 Functional substrate 501 Functional organic compound layer 502 Base material layer 503 Bonding layer 810 Solvent 900, 1100 Long film-like functional substrate

Claims (6)

A method for producing a cell culture container comprising a container part for containing cells and a medium,
The container part
A container body member;
A functional substrate comprising at least a base material layer and a functional organic compound layer disposed on one side of the base material layer;
The functional substrate is a bonding layer that exists between the base material layer and the surface of the container body member so that the functional organic compound layer faces the space for accommodating cells and culture medium on the surface of the container body member. Are joined via
The bonding layer is a layer composed of a material that does not substantially exhibit adhesiveness in a dry state and exhibits adhesiveness in a wet state with a solvent,
The following steps:
In the functional substrate in which the bonding layer in a dried state is further disposed on the other side of the base material layer, the bonding layer is wetted with a solvent, and the functional substrate is wetted with the bonding layer, A contact step of bringing the bonding layer and the surface into contact with the surface of the container main body member; and volatilizing the solvent while maintaining the contact state, whereby the functional substrate is formed on the surface of the container main body member. A method including a bonding step of bonding the substrate via the bonding layer. The contact process is
A step of preparing a functional substrate in a state where the bonding layer is wetted with a solvent by wetting the bonding layer with a solvent in the functional substrate in a state where the bonding layer is dried;
Preparing a container body member;
Contacting the functional substrate with the surface of the container body member. The contact process is
A step of preparing a functional substrate in a state where the bonding layer is dried and a container body member in a state where a solvent is present on the surface;
Contacting the functional substrate with the surface of the container body member. A method for producing a cell culture container comprising a container part for containing cells and a medium,
The container part
A container body member;
A functional substrate comprising at least a base material layer and a functional organic compound layer disposed on one side of the base material layer;
The functional substrate is a bonding layer that exists between the base material layer and the surface of the container body member so that the functional organic compound layer faces the space for accommodating cells and culture medium on the surface of the container body member. Are joined via
The bonding layer is a layer composed of a material that is weakly tacky or does not substantially exhibit tackiness in a dry state and exhibits tackiness in a solvent wet state;
The following steps:
In the container body member in which the bonding layer in a dried state is disposed on the surface, the bonding layer is wetted with a solvent, and the functional substrate is placed on the container body member in the state in which the bonding layer is wet. And a contact step in which the bonding layer comes into contact with each other, and
A method comprising a bonding step of bonding the functional substrate to the surface of the container body member via the bonding layer by volatilizing the solvent while maintaining a contact state. The functional substrate is a film-like functional substrate;
The long functional substrate wound in a roll shape with the bonding layer dried in advance is fed out, and the fed out functional substrate is cut into a shape to be bonded to the container body member. The method according to claim 1, further comprising a cutting step. The functional substrate is a film-like functional substrate;
The method further includes a cutting step of feeding out the long functional substrate wound in a roll shape prepared in advance and cutting the fed-out functional substrate into a shape to be joined to the container body member. 4. The method according to 4.

Images