JP5998464B2 - 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.

JP-A-2-21865 JP 2010-98799 A JP 2009-154904 A JP 2004-134020 A

  For example, Patent Document 3 and Patent Document 4 disclose a seal sticking device or the like for sticking seals to a desired product. A label applicator (labeler) that sucks and holds the surface of a label piece (functional substrate) as disclosed in Patent Document 3 and Patent Document 4 in order to fix a separately prepared functional substrate to the bottom of the container ).

  However, when the functional substrate is held by a labeler, the functional organic compound layer side of the functional substrate is sucked and held, so that the functional organic compound layer is partially destroyed by the labeler's suction force. There is also a risk that functions such as cell culture may be impaired. Even when the functional substrate is manually pasted, the contact with the functional organic compound layer increases in the pasting process, and functions such as cell culture may be similarly impaired.

  Accordingly, the present invention provides a method for producing a cell culture container, which can minimize the contact of another member with the functional organic compound layer when fixing the functional substrate and can maintain the function of the functional organic compound layer satisfactorily. It aims to provide a means for.

The present inventors use a functional base with a gripped portion provided with one or more gripped portions on the periphery of the functional base portion, and hold the functional base in a state where the gripped portion is gripped by a gripping tool or the like. It discovered that the said subject of maintaining the function of a functional organic compound layer favorably by joining to a main-body member was discovered. That is, the present invention includes the following inventions.
(1) A functional base part comprising at least an adhesive layer, a base material layer disposed on the adhesive layer, and a functional organic compound layer disposed on the base material layer, and a peripheral edge of the functional base part A functional substrate with a gripped portion, comprising one or more gripped portions provided.
(2) The function with the gripped portion of (1), wherein the gripped portion is formed by extending the base material layer from the peripheral edge of the functional base portion, and the gripped portion does not include the adhesive layer. Sex substrate.
(3) The functional substrate with a gripped portion according to (1) or (2), wherein the gripped portion is formed so as to be separable from the functional base portion.
(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 an adhesive layer, a base material layer disposed on the adhesive layer, and a functional organic compound layer disposed on the base material layer;
Having at least
The following steps:
(1) to (3), while holding the gripped portion of the functional base with gripped portion, the adhesive layer and the surface of the container body member are brought into contact with each other, and the functional base with gripped portion is A method comprising a joining step of joining to a surface of a container body member.
(5) The method according to (4), further comprising a separation step of separating the gripped portion from the functional base portion after the joining step.
(6) A method for producing a functional substrate with a gripped portion according to (2),
The following steps:
A base material in which a pressure-sensitive adhesive layer is disposed in a band-shaped region that is a part of one surface, and a functional organic compound layer is disposed on at least a part of the other surface including a portion located on the back side of the band-shaped region, A portion of each of the plurality of partial regions having different lengthwise positions in the belt-shaped region is a functional base portion, and is adhered to the one surface side adjacent to each of the portions. A method comprising a cutting step of forming a plurality of functional substrates with gripped portions by cutting so that at least one portion where no layer is disposed becomes a gripped portion.
(7) The base material is a long film base material,
The band-shaped region is a band-shaped region along the length direction of the long film substrate.
Method (6).
(8) A cell culture container provided with a container part for containing cells and a medium,
The container part
A container body member;
Provided at the periphery of the functional substrate portion, the functional substrate portion comprising at least an adhesive layer, a base material layer disposed on the adhesive layer, and a functional organic compound layer disposed on the base material layer A functional substrate with a gripped portion, including one or more gripped portions;
A cell culture vessel having at least

  According to the present invention, since the container body member and the functional base can be joined while gripping the gripped portion, it is possible to avoid contact of other members, tools, and the like with the functional organic compound layer. And does not impair the function of the functional organic compound.

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 a functional base | substrate. A is a top view of an example of a functional substrate with a gripped portion. B is a cross-sectional view taken along line I-I 'of A. FIG. C is a sectional view taken along line II-II ′ of A. FIG. A is a top view of an example of a functional substrate with a gripped portion. B is a cross-sectional view taken along line I-I 'of A. FIG. C is a sectional view taken along line II-II ′ of A. FIG. A is a top view of an example of a functional substrate with a gripped portion. B is a cross-sectional view taken along line I-I 'of A. FIG. C is a sectional view taken along line II-II ′ of A. FIG. A is a top view of an example of a functional substrate with a gripped portion. B is a cross-sectional view taken along line I-I 'of A. FIG. C is a sectional view taken along line II-II ′ of A. FIG. A is a top view of an example of a functional substrate with a gripped portion. B is a cross-sectional view taken along line I-I 'of A. FIG. C is a sectional view taken along line II-II ′ of A. FIG. A is a top view of an example of a functional substrate with a gripped portion. B is a cross-sectional view taken along line I-I 'of A. FIG. C is a sectional view taken along line II-II ′ of A. FIG. A is a plan view of an example 1200, 1200 ', 1200' 'of a functional substrate with a gripped portion. B is a cross-sectional view taken along the line III-III ′ of the functional substrate 1200 with a gripped portion shown in A. FIG. C is a cross-sectional view taken along the line III-III ′ of the functional base body 1200 ′ with a gripped portion shown in FIG. D is a cross-sectional view taken along the line III-III ′ of the functional base body 1200 ″ with the gripped portion shown in FIG. It is a perspective view for demonstrating one Embodiment of the manufacturing method of the cell culture container of this invention. The next process of FIG. 13A is shown. The next process of FIG. 13B is shown. It is a perspective view for demonstrating one Embodiment of the manufacturing method of the cell culture container of this invention. The next process of FIG. 14A is shown. The next process of FIG. 14B is shown. It is a perspective view for demonstrating one Embodiment of the manufacturing method of the cell culture container of this invention. The next process of FIG. 15A is shown. A is a top view of one embodiment of a half-cut long laminate. B is a sectional view taken along line IV-IV ′ of A. FIG. C is a cross-sectional view taken along line V-V ′ of A. FIG. A is a top view of one embodiment of a half-cut long laminate. B is a sectional view taken along line IV-IV ′ of A. FIG. C is a cross-sectional view taken along line V-V ′ of A. FIG. A is a top view of one embodiment of a half-cut long laminate. B is a sectional view taken along line IV-IV ′ of A. FIG. C is a cross-sectional view taken along line V-V ′ of A. FIG. It is a figure for demonstrating the strip | belt-shaped area | region of the base-material surface.

  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, either the joining process of the container body member 109 and the side wall member 111 or 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).

  The functional substrates 140 and 210 include an adhesive layer 503 including an adhesive or an adhesive, a base material layer 502 disposed on the adhesive layer 503, and a functional organic compound layer 501 disposed on the base material layer 502. Are attached to the bottom portions 101 and 201 of the container body member or the flat container body member 109 shown in FIG.

<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>
The present invention is characterized in that a cell culture container having a functional substrate is produced using a functional substrate with a gripped part. For convenience of explanation, the functional substrate finally fixed to the cell culture container is provided. The structure of will be described first. “Functional substrate” in the “functional substrate with gripped portion” is a term for clarifying that the functional substrate is included as a part, except that a gripped portion is provided at the periphery. The functional substrate has the following characteristics.

  As shown in FIG. 5, the cross section along the thickness direction of the functional substrates 140 and 210 in the present invention is an adhesive layer 503, a base material layer 502 disposed on the adhesive layer 503, and the base material layer 502. At least a functional organic compound layer 501 disposed thereon is provided. 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 adhesive layer 503 and 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.

<Adhesive layer>
The pressure-sensitive adhesive layer 503 includes an adhesive or a pressure-sensitive adhesive. The adhesive and the pressure-sensitive adhesive are not particularly limited as long as they can join the base layer of the functional substrate and the container main body member, respectively. Specifically, the adhesive includes a polyacrylate adhesive, Cyanoacrylate adhesives, polyester adhesives, polyamide adhesives, polyimide adhesives, epoxy adhesives, polyurethane adhesives, methacrylic adhesives, rubber adhesives, silicone adhesives, inorganic adhesives Examples of such adhesives include acrylic adhesives, urethane adhesives, silicone adhesives, and rubber adhesives. These adhesives or pressure-sensitive adhesives are applied on the base material layer 502 to form the pressure-sensitive adhesive layer 503.

<Functional substrate with gripped part>
In the present invention, when the functional substrate is bonded to the container body member, a “functional substrate with a gripped portion” in which a gripped portion is further provided on the periphery of the functional substrate is used. A portion corresponding to the functional substrate in the functional substrate with the gripped portion is referred to as a “functional substrate portion”. The functional base portion has the above-described features described for the functional base.

A specific example of the functional base with the gripped portion will be described with reference to FIGS.
A functional base 600 with a gripped portion shown in FIG. 6 includes a rectangular functional base portion 601 and four gripped portions 602 provided on the periphery thereof. The number, shape, and dimensions of the gripped portions are not particularly limited, and can be appropriately selected according to the shape of the gripping tool, the shape of the functional base portion, the shape of the container body member to be joined, and the like. The number of gripped portions 602 is not limited to four, and may be at least one.

  The gripped portion 602 is preferably formed by extending the base material layer 502 from the periphery of the functional base portion 601. The gripped portion 602 can be formed by extending not only the base material layer 502 but also the functional organic compound layer 501 and the adhesive layer 503 (FIGS. 6B and 6C).

  When the gripped portion 602 including the adhesive layer 503 is gripped by a gripping tool, the adhesive material or the adhesive constituting the adhesive layer 503 may adhere to the contact portion of the tool. This may not be desirable when the gripping portion 602 is repeatedly gripped using the same gripping tool. The problem is that the gripping tool that grips the gripped portion 602 including the adhesive layer 503 avoids adhesion by releasing the contact surface with the adhesive layer 503, or the gripping tool including the contact surface Although it can be solved by measures such as configuring the member to be replaceable, it can be solved more reliably by making the surface of the gripped portion not include the adhesive layer 503.

  For example, in the functional substrate 700 with the gripped portion shown in FIG. 7, the gripped portion 702 does not include the adhesive layer 503, and the base layer 502 and the functional organic compound layer 501 extending from the periphery of the functional base portion 701 are included. Including.

  In the functional substrate 800 with a gripped portion shown in FIG. 8 which is another preferred form, the gripped portion 802 does not include the adhesive layer 503 and the functional organic compound layer 501, and extends from the periphery of the functional base portion 801. A substrate layer 502 is included.

  6 to 8, the functional base portions 601, 701, and 801 include a functional organic compound layer 501, a base material layer 502, and an adhesive layer 503.

  The functional base portions 601, 701, 801 and the gripped portions 602, 702, 802 can be formed continuously and integrally, but the gripped portions 602, 702, 802 are separated after joining to the container body member. For this embodiment, a perforation, a half cut line (a cut partially cut in the thickness direction), or the like can be provided at the boundary portion between the two. 6 to 8 show examples in which perforations 610, 710, and 810 are provided. However, even in the embodiment in which the gripped portion is separated from the functional base portion, a perforation or the like is unnecessary when the boundary portion between the two is cut by a cutting means such as scissors.

  The shape of the functional substrate portion is not particularly limited as described above with respect to the functional substrate, and can be an appropriate shape according to the shape of the container body member. For example, in order to obtain the functional substrate 140 to be bonded to the bottom surface of the flask-type container shown in FIG. 1A and the like, as shown in FIGS. 9, 10, and 11, the rectangular portion and its one side are used as the lower side. Functional bases 900, 1000, 1100 with gripped portions including functional base portions 901, 1001, 1101 having a shape in which trapezoidal portions are continuously provided and gripped portions 902, 1002, 1102 provided at the periphery are used. be able to. Regarding the layer structure of the functional base portions 901, 1001, 1101 and the gripped portions 902, 1002, 1102, the functional base portions 601, 701, 801 and the gripped portion 602 described with reference to FIGS. , 702 and 802, the description is omitted.

  FIG. 12 shows functional bases 1200, 1200 ′, and 1200 ″ with gripped portions in which the functional base portion 1201 is circular and the gripped portions 1202, 1202 ′, and 1202 ″ are one. Also in these embodiments, the gripped portions 1202, 1202 ′, 1202 ″ are formed by partially extending the base material layer 502 from the peripheral edge of the functional base portion 1201. The functional base 1201 includes a functional organic compound layer 501, a base material layer 502, and an adhesive layer 503. The gripped portion 1202 includes a functional organic compound layer 501 and an adhesive layer 503 (FIG. 12B), and the gripped portion 1202 ′ does not include the adhesive layer 503 and includes the functional organic compound layer 501 (FIG. 12C). 1202 '' does not include the functional organic compound layer 501 and the adhesive layer 503 (FIG. 12B).

<Method for producing cell culture container>
A method for producing a cell culture container using the above-described functional base with a gripped portion will be described with reference to FIGS.

  FIGS. 13A to 13C are examples in which the flask-type container 100 shown in FIG. 1A and the like is configured using the functional base 900 with a gripped portion.

  The gripped portion 902 of the functional base 900 with the gripped portion is gripped (FIG. 13A). Although gripping may be performed with the fingertip of the operator, it is more preferable to perform gripping using a gripping tool 1301 such as tweezers as illustrated. By grasping the grasped portion 902, the functional organic compound layer 501 on the surface of the functional base portion 901 can be prevented from coming into contact with the grasping tool 1301 or the like. Physical damage can be avoided. While maintaining the gripping state, the adhesive layer 503 of the functional base portion 901 and the surface (bottom surface) of the container main body member 103 are brought into contact with each other, and the functional base 900 with the gripping portion is joined to the surface of the container main body member 130 (FIG. 13A, B). The gripped portion 902 stands up so as to form an appropriate angle with respect to the functional base portion 901 as necessary, as shown in FIG. 13A, etc., by bending the boundary portion with the functional base portion 901. be able to. Although not shown, when the functional substrate 900 with the gripped portion is joined to the surface of the container body member opened to the side, the flat shape as shown in FIG. It is also possible to carry out the joining step in a state where the tool is brought close and the gripped portion 902 is gripped.

  As shown in FIG. 13C, the gripped portion 902 is preferably separated from the functional base portion 901 after the joining step. The functional base 901 after separating the gripped portion 902 corresponds to the functional base 140. However, separation is not necessary when the size of the gripped portion 902 is very small. For applications where the functional substrate is exfoliated afterwards (for example, for exfoliating and collecting the functional substrate part to which the cells have adhered after cell culture) It is preferable to leave 902 as it is.

  The top surface member 104 is joined to the upper end of the container main body member 103 (FIG. 13C), and the flask-type container part 100 to which the functional substrate 140 is joined is completed.

  Other embodiment of the manufacturing method of the flask type container part 100 with which the functional base | substrate 140 was joined is shown to FIG. In this embodiment, the adhesive base 503 is brought into contact with the functional base 900 with a gripped portion on the surface of the flat container body member 109 while the gripped portion 902 is gripped with the gripping tool 1301, and both are bonded. (FIG. 14A). The container body member 109 to which the functional base 140 is fixed is obtained by appropriately separating the gripped portion 902 (FIG. 14B). Finally, the side wall member 111 and the top surface member 104 are joined (FIG. 14C), and the flask-type container unit 100 to which the functional substrate 140 is joined is completed.

  FIGS. 13A to 14C show an example in which the flask-type container unit 100 is manufactured using the functional substrate 900 with a gripped portion. However, when the functional substrates 1000 and 1100 with a gripped portion shown in FIGS. Even if it exists, it is possible to manufacture the flask type container part 100 similarly.

  FIG. 15 shows an embodiment of a method for manufacturing the dish-shaped container 200 to which the functional substrate 210 is bonded. In this embodiment, the adhesive body 503 on the surface of the functional base 1201 is brought into contact with the bottom surface of the container body member 203 while the functional base 1200 with a gripped part is held and the gripped part 1202 is gripped by the gripping tool 1301. Both are joined (FIG. 15A). The container body member 200 to which the functional base 210 (1201) is fixed is obtained by appropriately separating the gripped portion 1202 (FIG. 15B). Functional bases 1200 ′ and 1200 ″ with gripped portions that differ only in the structure of the gripped portion can be used in the same manner.

<Method for manufacturing functional substrate with gripped portion>
The manufacturing method of the functional substrate with the gripped portion is not particularly limited.
A functional base with a gripped portion having a gripped portion formed by extending a base material layer from the peripheral edge of the functional base portion has an adhesive layer disposed on one surface of a base material of an appropriate shape. It can be easily manufactured by disposing a functional organic compound layer on the other surface.

  In order to efficiently produce a functional substrate with a gripped portion, a long-sized substrate having an adhesive layer disposed on one surface and a functional organic compound layer disposed on the other surface It is preferable to prepare a plurality of functional substrates with gripped portions having the same shape by cutting the substrate into a predetermined shape.

  Based on FIG. 16, an example of a half-cut long laminate in which a plurality of regions corresponding to the functional base 600 with a gripped part are defined by half-cut lines will be described. A half-cut long laminate 1600 is a functional substrate with a gripped portion on a long laminate comprising a release film 1601, an adhesive layer 503, a base material layer 502, and a functional organic compound layer 501. A half-cut line 1610 that defines a plurality of regions corresponding to 600 is formed. The half-cut line 1610 is formed from the surface on the functional organic compound layer 501 side of the laminate to a thickness direction position that cuts the adhesive layer 503 in the thickness direction but does not completely cut the release film 1601. It is a score line. In the present invention, the term “cut” and the term “cut process” include a process of forming a half-cut line. The base material layer 502 is preferably composed of a film base material, and is preferably a long laminated film in which the entire half-cut long laminated body 1600 can be rolled up. A large number of functional substrates 600 with gripped portions can be obtained from the half-cut long laminate 1600.

  FIG. 17 shows an example of a half-cut elongated laminate in which a plurality of regions corresponding to the functional base 700 with a gripped part are defined by half-cut lines. A long laminate 1700 that has been half-cut has an adhesive layer 503 arranged on one surface of a long base material 502 in two band-like regions 1901 and 1902 shown in FIG. A half-cut line that defines a plurality of regions corresponding to the functional substrate 700 with a gripped portion in a long laminate in which the functional organic compound layer 501 is disposed throughout and the adhesive layer 503 is protected by the release film 1601. 1710 is formed. In FIG. 17, the region where the adhesive layer 503 does not exist and the gripped portion 702 does not exist is depicted so that only the release film 1601 remains, but the substrate is not bonded to the region via the adhesive layer 503. 502 may remain. According to the printing technique, it is easy to selectively provide the adhesive layer 503 only in the belt-like regions 1901 and 1902 on the surface of the long substrate 502. The long substrate 502 is particularly preferably a film substrate. The portions corresponding to each of the plurality of partial regions having different lengthwise positions in the band-shaped regions 1901 and 1902 become the functional base portion 701, and the adhesive layer 503 is not disposed on the surface adjacent to each of the portions 4 By forming the half-cut line 1710 so that one part becomes the gripped portion 702, the gripped portion 702 has no adhesive layer, and the functional base portion 701 has the gripped portion 503. The functional substrate 700 can be manufactured efficiently.

  FIG. 18 shows an example of a half-cut long laminate in which a plurality of regions corresponding to the functional base 800 with a gripped portion are defined by half-cut lines. A long laminate 1800 that has been half-cut has an adhesive layer 503 disposed on one surface of a long base material 502 in two band-like regions 1901 and 1902 shown in FIG. The functional organic compound layer 501 is disposed only on the back side positions of the band-like regions 1901 and 1902 and the adhesive layer 503 is protected by the release film 1601 to correspond to the functional substrate 800 with a gripped portion. A half-cut line 1810 that defines a plurality of regions is formed. According to this embodiment, a function with a gripped portion in which the adhesive layer and the functional organic compound layer are not present in the gripped portion 802 and the adhesive layer 503 and the functional organic compound layer 501 are present in the functional base portion 801. The conductive substrate 800 can be efficiently manufactured.

  The number of belt-like regions is not limited to two, and may be one or may be three or more.

  In the long laminated bodies 1600, 1700, and 1800 that have been half-cut, perforations or the like that further facilitate separation are provided at the boundaries between the functional base portions 601, 701, and 801 and the gripped portions 602, 702, and 802. Alternatively, a perforation or the like may be provided at the boundary after separation from the release film 1610.

100, 200 Container part 101, 201 Bottom part 102, 202 Side wall part 103, 109, 203 Container body member 104 Top surface member 111 Side wall member 130, 220 Space 140, 210 Functional substrate 600, 700, 800, 900, 1000, 1100 1200, 1200 ′, 1200 ″ functional substrate with gripped portion 601, 701, 801, 901, 1001, 1101, 1201 Functional substrate portion 602, 702, 802, 902, 1002, 1102, 1202, 1202 ′, 1202 '' to-be-held portion 610, 710, 810, 910, 1010, 1110, 1210 perforation 1301 gripping tool 1600, 1700, 1800 half-cut long laminate 1901, 1902

Claims (5)

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 an adhesive layer, a base material layer disposed on the adhesive layer, and a functional organic compound layer disposed on the base material layer;
Having at least
The following steps:
Provided at the periphery of the functional substrate portion, the functional substrate portion comprising at least an adhesive layer, a base material layer disposed on the adhesive layer, and a functional organic compound layer disposed on the base material layer While holding the gripped portion of the functional substrate with the gripped portion provided with one or more gripped portions, the adhesive layer and the surface of the container body member are brought into contact with each other, and the functional substrate with the gripped portion is placed in the container. Bonding process for bonding to the surface of the body member
Including
After the bonding step further comprises a separation step of separating the gripped portion of the functional base portion, Methods. The method according to claim 1, wherein the gripped portion is formed by extending the base material layer from the peripheral edge of the functional base portion, and the gripped portion does not include an adhesive layer. The method according to claim 1 or 2, wherein the gripped portion is formed so as to be separable from the functional base portion. Provided at the periphery of the functional substrate portion, the functional substrate portion comprising at least an adhesive layer, a base material layer disposed on the adhesive layer, and a functional organic compound layer disposed on the base material layer One or more gripped portions, the gripped portion is formed by extending a base material layer from the peripheral edge of the functional base portion, and the gripped portion does not include an adhesive layer. A method for producing a functional substrate with a part, comprising:
The following steps:
A base material in which a pressure-sensitive adhesive layer is disposed in a band-shaped region that is a part of one surface, and a functional organic compound layer is disposed on at least a part of the other surface including a portion located on the back side of the band-shaped region, A portion of each of the plurality of partial regions having different lengthwise positions in the belt-shaped region is a functional base portion, and is adhered to the one surface side adjacent to each of the portions. A method comprising a cutting step of forming a plurality of functional substrates with gripped portions by cutting so that at least one portion where no layer is disposed becomes a gripped portion. The substrate is a long film substrate,
The band-shaped region is a band-shaped region along the length direction of the long film substrate.
The method of claim 4 .

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