JP2013099286A - Cell culture container - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cell culture container having a functional substrate body designed to reduce the interference of an adhesive or a sticky agent against the function of the cell culture container required on the observation of cells or the like.SOLUTION: The cell culture container 700 is characterized in that a container portion has at least a container body member (201', 202) and a functional substrate body 210; the peripheral edge region of the substrate layer 502 side surface of the functional substrate 210 is bound to the container body member surface region facing the peripheral edge region through the adhesive or a sticky agent 750; and the adhesive or sticky agent does not exit in the central region of the substrate layer 502 side surface. Preferably, an opening 701 is formed in a portion surrounded by a portion adhered to the peripheral edge region of the surface of the container body member on substrate layer 502 side of the functional substrate 210, and the central region of the substrate layer 502 side surface is exposed through the opening 701.

Description

  The present invention relates to a cell culture vessel 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, on which a functional organic compound layer such as a temperature-responsive polymer is coated. 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. There is still room for improvement.

  Therefore, in Patent Document 2 and the like, a technique is proposed in which a functional substrate having a functional compound layer on the surface and having a film-like or plate-like shape is separately prepared and the functional substrate is fixed to a cell culture vessel. ing. 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

  In the prior art disclosed in Patent Document 2 and the like for fixing a functional substrate to a cell culture container, the entire surface of the functional substrate on the base material layer side and the surface of the container body member are bonded with an adhesive or a pressure-sensitive adhesive. It is usual to join via the etc. (for example, patent document 2 FIG. 1).

  However, when the adhesive or the pressure-sensitive adhesive is present on the entire surface of the functional substrate on the base layer side, various disadvantages occur. For example, when a functional substrate is bonded to a dish-shaped cell culture container having an opening at the center of the bottom plate so as to close the opening from the inside of the container with the substrate layer side surface facing down, If the adhesive or pressure-sensitive adhesive is present on the entire surface of the layer side, dust or the like adheres to the adhesive or pressure-sensitive adhesive on the base layer side surface exposed from the opening, which hinders observation of cultured cells. There's a problem. Furthermore, even when a functional substrate is adhered to a cell culture container having a normal bottom plate member having no opening with the substrate layer side surface facing down, the entire surface of the substrate layer side surface If the adhesive or pressure sensitive adhesive is present in the cell, when the cultured cells in the container are observed with a fluorescence microscope, the autofluorescence due to the adhesive or pressure sensitive adhesive on the lower surface of the central part of the functional substrate becomes a noise and obstructs the observation. There is.

  Accordingly, an object of the present invention is to provide a cell culture container provided with a functional substrate, which is designed to reduce interference by an adhesive or an adhesive with respect to the function of the cell culture container required at the time of cell observation or the like. .

  The present inventors solve the above-mentioned problem by bonding the functional substrate and the container body member by disposing an adhesive or a pressure-sensitive adhesive only in a partial region on the base material layer side surface of the functional substrate. I found out. The present invention includes the following inventions.

(1) 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 the base material layer;
Comprising at least
The surface of the functional substrate on the base material layer side is fixed to the container body member so that the surface of the functional organic compound layer side faces the space for accommodating cells and culture medium. A part of the region and a region of the surface of the container main body member facing the part of the part are bonded via an adhesive or a pressure-sensitive adhesive,
No adhesive or pressure-sensitive adhesive is present on a region other than the partial region on the surface of the functional substrate on the base material layer side,
A cell culture container characterized by the above.
(2) The partial region of the surface of the functional substrate on the substrate layer side is a peripheral region of the surface of the functional substrate on the substrate layer side, and the surface of the functional substrate on the substrate layer side The cell culture vessel according to (1), wherein an adhesive and an adhesive are not present on the region surrounded by the peripheral region.
(3) an opening is formed in a portion surrounded by a peripheral region of the surface of the container body member on the base material layer side of the functional substrate and a portion where the surface is bonded via an adhesive or an adhesive, The cell culture container according to (2), wherein a region surrounded by the peripheral region on the surface of the functional substrate on the base material layer side is exposed through the opening.

  In the cell culture container provided with the functional substrate according to the present invention, the function of the cell culture container is less disturbed by the adhesive or the adhesive.

FIG. 1A shows an example of the cell culture container of the present invention. FIG. 1B is a cross-sectional view taken along the line I-I ′. FIG. 1C is a cross-sectional view taken along the line II-II ′. FIG. 2A shows another example of the cell culture container of the present invention. FIG. 2B is a cross-sectional view taken along the line III-III ′. FIG. 3 shows an example of the case where the container portion is completed by combining other members after the functional substrate is fixed to the container partial member. FIG. 4 shows another example in the case where the container portion is completed by combining other members after the functional substrate is fixed to the container partial member. FIG. 5 shows a cross-sectional view of the functional substrate. FIG. 6A shows an example of the cell culture container of the present invention in which an opening is formed in a part of the container. FIG. 6B is a cross-sectional view taken along the line IV-IV ′. FIG. 7A shows another example of the cell culture container of the present invention in which an opening is formed in a part of the container. FIG. 7B is a V-V ′ sectional view.

<Shape of cell culture vessel>
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 FIG. 1, the container portion may have a shape in which a space for containing cells and culture medium is closed by a wall surface, or a shape in which one end of the space is opened as shown in FIG. May be.

  An example of a preferred embodiment of the container portion is shown in FIG. 1A includes a bottom portion 101, a side wall portion 102 standing on the periphery of the bottom portion 101, and a top surface portion 103 that is joined to the upper end portion of the side wall portion 102 and is disposed to face the bottom portion 101. At least. A through-hole 104 is formed in a part of the side wall portion 102, and a container portion having a shape called a “flask type” including a neck portion 105 extending from the periphery of the through-hole 104 to the outside of the container portion. A locking portion 106 for locking the lid 110 is formed on the neck portion 105 of the container portion 100, and the lid 110 is detachably mounted through the locking portion. 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 portion 100, and FIG. 1C shows a cross-sectional view taken along the line II-II ′. A space 130 for accommodating cells and a culture medium is formed in a portion of the container 100 surrounded by the bottom 101, the side wall 102, and the top surface 103. A functional substrate 140 is fixed to a part of the inner wall surface facing the space 130 (the bottom 101 in the embodiment shown in FIG. 1).

  As other embodiment of a container part, as shown in FIG. 2, the dish-shaped container part 200 provided with the bottom part 201 and the side wall part 202 standingly arranged in the periphery of the bottom part 201 is mentioned. A functional substrate 210 is fixed to the surface (inner bottom surface) of the bottom portion 201 on the side of the space 220 for accommodating cells and culture medium.

  A partial region of the surface of the functional substrate on the side of the base material layer and a region of the bottom surface facing the region are bonded via an adhesive or pressure-sensitive adhesive 150 or 250.

  In the present invention, a portion of the container portion excluding the functional substrate is referred to as a “container body member”. For example, in the container part 100 of FIG. 1, the container body member is a part composed of the bottom part 101, the side wall part 102, the top surface part 103 and the neck part 105. In the container part 200 of FIG. 2, the container body member is It is a part composed of a bottom part 201 and a side wall part 202.

  In order to manufacture a container body member to which a functional substrate is bonded, the functional substrate may be bonded to the container body member, or a member that is a part of the container body member (hereinafter referred to as “container partial member”). In some cases, the functional base may be bonded to the container part, and then the container part member and another member may be bonded to complete the container part. The point of dividing the container portion in this way will be described in more detail with reference to FIGS.

  As shown in FIG. 3, in one embodiment for forming the container portion 100 after bonding the functional substrates, first, the functional substrate 140 is placed on the inner bottom surface of the container portion member 301 composed of the bottom portion 101 and the side wall portion 102. Join. Subsequently, the container part 100 is formed by joining the top surface member 302 corresponding to the top surface part 103. The container part member 301 and the top surface member 302 are joined in a liquid-tight manner so that the culture solution does not leak out according to the purpose of cell culture.

In another embodiment shown in FIG. 4 for forming the container part 100 after bonding of the functional substrates, first, the functionality of the container part member 401 corresponding to the bottom part 101 on the surface facing the space 130 after completion is provided. The substrate 140 is bonded. The container part 100 is formed by joining the side wall member 402 corresponding to the side wall part 102 provided with the neck part 105 and the top surface member 302 corresponding to the top surface part 103.
The combination of a plurality of members is not limited to the form shown in FIGS.

<Material of cell culture container>
The material which forms the member of other cell culture containers, such as a container main body member and a lid | cover, is not specifically limited, The material 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 layer 502 and a functional organic compound layer 501 disposed on the base layer 502. . 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 is typically a transparent functional substrate.
The functional substrate can be produced by forming a functional organic compound layer on the substrate layer by an appropriate method. One or more other layers (for example, a primer layer described later) may be present between the base material layer and the functional organic compound layer 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, the shape 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 substrate layer 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 should just contain the material which can form the above-mentioned functional organic compound layer in one surface, and the kind of material is not specifically 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), vinyl chloride, vinylidene chloride, polyphenylene sulfide, polyether sulfone, polyethylene naphthalate, polypropylene, acrylic, and other inorganic materials such as glass and quartz. preferable.

  The surface of the base material layer on the side on which the functional organic compound layer is formed can be a surface 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 can be appropriately selected. 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 is not particularly limited as long as it has a desired function, but more preferably it can be changed from cell adhesiveness to cell nonadhesiveness by a predetermined stimulus. And a hydrophilic compound such as an ethylene glycol chain composed of one or more ethylene glycol units (CH ₂ —CH ₂ —O).

  The film thickness of the functional organic compound layer may be, for example, in the range of 0.5 nm to 300 nm, and 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 is particularly preferably a stimulus-responsive polymer layer. The stimulus-responsive polymer layer is a layer containing a polymer in which the degree of cell adhesion on the surface 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-mentioned 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.

  Bio-related substances that can be covalently bonded to the end of 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.

<Bonding of functional substrate and container body member>
In the present invention, the base layer of the functional base is fixed to the container body member in such a manner that the surface on the side of the functional organic compound layer faces the space for accommodating cells and culture medium. A partial region of the side surface and a region of the surface of the container body member facing the partial region are bonded via an adhesive or a pressure-sensitive adhesive, and the functional substrate base An adhesive and a pressure-sensitive adhesive are not present on a region other than the partial region on the surface on the material layer side.

  In the present invention, 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 part. Specifically, the adhesive is polyacrylic acid. Ester adhesives, cyanoacrylate adhesives, polyester adhesives, polyamide adhesives, polyimide adhesives, epoxy adhesives, polyurethane adhesives, methacrylic adhesives, rubber adhesives, silicone adhesives An adhesive such as an adhesive or an inorganic adhesive can be exemplified, and examples of the adhesive include an acrylic adhesive, a urethane adhesive, a silicone adhesive, and a rubber adhesive.

  In the present invention, a partial region of the surface of the functional substrate on the base material layer side and a region of the surface of the container main body member facing the partial region are bonded via an adhesive or a pressure-sensitive adhesive. And an adhesive and a pressure-sensitive adhesive are not present on a region other than the partial region on the surface of the functional substrate on the base material layer side.

  Here, the “partial region” is not particularly limited, but is preferably a peripheral region of the surface on the base material layer side in a sheet-like functional substrate. At this time, the adhesive and the pressure-sensitive adhesive are not present in the region (central region) of the surface on the base material layer side surrounded by the peripheral region.

  Specifically, in the flask-type cell culture container shown in FIG. 1, the periphery of the surface (lower surface) of the functional substrate 140 on the base material layer 502 side that covers substantially the entire inner surface of the container of the bottom 101 in the container unit 100. The region and the surface region of the bottom 101 facing the region are joined via an adhesive or pressure-sensitive adhesive layer 150. No adhesive or pressure-sensitive adhesive is present in the central region surrounded by the peripheral region on the surface of the functional substrate 140 on the base material layer 502 side. Similarly, also in the dish-type cell culture container shown in FIG. 2, the peripheral edge of the surface (lower surface) of the functional substrate 210 on the base material layer 502 side that covers substantially the entire inner surface of the bottom 201 in the container 200. The region and the surface region of the bottom 201 facing the region are joined via an adhesive or pressure-sensitive adhesive layer 250. No adhesive or pressure-sensitive adhesive is present in the central region surrounded by the peripheral region on the surface of the functional substrate 210 on the base material layer 502 side. With the configuration illustrated in FIGS. 1 and 2, a failure caused by an adhesive or a pressure-sensitive adhesive in a central region mainly observed when observing cultured cells (for example, an autogenous caused by the adhesive or the pressure-sensitive adhesive at the time of observation with a fluorescence microscope). It is possible to perform highly accurate observation without fluorescence. The adhesive or adhesive layers 150 and 250 provided between the peripheral region of the surface of the functional substrate 140 and 210 on the base material layer 502 side and the bottom portions 101 and 201 are arranged in the circumferential direction of the functional substrate 140 and 210. It may be formed continuously along the entire circumference along the circumference, or may be formed intermittently along the circumferential direction of the functional bases 140 and 210.

  In FIGS. 1 and 2, for the sake of explanation, the thickness of the adhesive or pressure-sensitive adhesive layers 150, 250 is emphasized, and the surface of the functional substrate 140, 210 on the substrate layer 502 side is depicted. It is depicted that a gap is formed between the central region and the opposing surfaces of the bottom portions 101 and 201. However, when the adhesive or pressure-sensitive adhesive layers 150 and 250 are very thin layers compared to the functional substrates 140 and 210, or when the functional substrates 140 and 210 have flexibility, the The central region of the surface of the conductive bases 140 and 210 on the base material layer 502 side and the surfaces of the bottoms 101 and 201 are in contact with each other.

  In another embodiment of the present invention, as shown in FIGS. 6 and 7, the surface of the container body member is bonded to the peripheral region of the surface of the functional substrate on the base material layer side via an adhesive or an adhesive. An opening is formed in a portion surrounded by the portion. At this time, the region surrounded by the peripheral region of the surface of the functional substrate on the base material layer side is exposed through the opening.

  A flask-type container according to an embodiment having an opening includes, for example, as shown in FIG. 6A, a bottom 101 ′ having a peripheral portion surrounding the opening 601 in which an opening 601 penetrating in the thickness direction is formed in the center. It can be manufactured by fixing the functional base 140 to a container partial member 600 provided with a side wall portion 102 erected on the periphery of the bottom portion 101 ′ and combining the top surface member 302 corresponding to the top surface portion 103. FIG. 6B shows a cross-sectional view of the intermediate product in a state where the functional base 140 is fixed to the container partial member 600. At this time, the peripheral region of the surface of the functional substrate 140 on the side of the base material layer 502 and the surface of the peripheral portion of the bottom 101 ′ facing the region are bonded via the adhesive or pressure-sensitive adhesive layer 650. . The central region of the surface of the functional base 140 on the base material layer 502 side is exposed outside the container through the opening 601. Since the adhesive and the pressure-sensitive adhesive are not present on the exposed surface of the base material layer 502, problems such as adhesion of dust can be avoided. A cell culture container having an opening formed at the center of the bottom and having the opening covered with a functional substrate has the advantage that bubbles do not enter between the functional substrate 140 and the bottom.

  As shown in FIG. 7A, for example, the dish-shaped cell culture container according to the embodiment having an opening has a bottom portion 201 ′ having a peripheral portion surrounding the opening 701 in which an opening 701 penetrating in the thickness direction is formed at the center. And a container portion 700 including a side wall portion 202 erected on the periphery of the bottom portion 201 ′. FIG. 7B shows a cross-sectional view of the container part 700. The peripheral region on the surface of the functional substrate 210 on the base material layer 502 side and the peripheral region on the surface of the bottom 201 ′ are bonded via an adhesive or pressure-sensitive adhesive layer 750. A central region of the surface of the functional base 210 on the base layer 502 side is exposed outside the container through the opening 701. There is no adhesive or pressure-sensitive adhesive on the exposed surface of the base material layer 502.

  In the present invention, the procedure for joining the functional substrate and the container main body member or the container partial member with an adhesive or a pressure-sensitive adhesive is not particularly limited. (1) Pre-adhering to a predetermined partial region on the base material layer side surface Preparing a functional substrate on which a layer of an agent or an adhesive is formed, and bonding the functional substrate to a container body member or a container partial member; (2) Adhesive or adhesive in advance on a predetermined partial region of the surface A container body member or a container part member on which an agent layer is formed is prepared, and the functional substrate and the container body member or the container part member are joined; (3) the functional substrate and the container body member or the container part member; And applying adhesive or pressure-sensitive adhesive to at least one predetermined region of the surface of the functional substrate on the base material layer side and the surface of the container body member or the container partial member, May be joined by any procedure such as joining

  The functional organic compound layer is also used in the case of manufacturing a functional substrate in which an adhesive or pressure-sensitive adhesive layer is previously formed in a predetermined partial region on the base material layer side surface used in (1) above. The order of forming the layer and the layer of the adhesive or pressure-sensitive adhesive is not limited. For example, (4) a base material in which an adhesive or pressure-sensitive adhesive layer is formed in a predetermined partial region, and the surface of the adhesive or pressure-sensitive adhesive layer is protected by a release sheet, if necessary, is prepared. The functional organic compound layer may be laminated on the surface on which the adhesive or pressure-sensitive adhesive layer is not formed, or (5) after the functional organic compound layer is laminated on one surface of the substrate The adhesive or pressure-sensitive adhesive layer may be formed in a predetermined partial region on the other surface of the substrate, and the surface of the adhesive or pressure-sensitive adhesive layer may be protected as necessary. The “base material in which an adhesive or pressure-sensitive adhesive layer is formed in a predetermined partial region and the surface of the adhesive or pressure-sensitive adhesive layer is protected by a release sheet as necessary” used in (4) above is uniquely You may manufacture and may purchase a commercial item.

100, 200... Container portions 101 to 106, 201 to 202... Container body members 150, 250, 650, 750... Adhesives or adhesives 140, 210. Material layer 501... Functional organic compound layers 601, 701... Opening

Claims (3)

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 the base material layer;
Comprising at least
The surface of the functional substrate on the base material layer side is fixed to the container body member so that the surface of the functional organic compound layer side faces the space for accommodating cells and culture medium. A part of the region and a region of the surface of the container main body member facing the part of the part are bonded via an adhesive or a pressure-sensitive adhesive,
No adhesive or pressure-sensitive adhesive is present on a region other than the partial region on the surface of the functional substrate on the base material layer side,
A cell culture container characterized by the above.   The partial region of the surface of the functional substrate on the substrate layer side is a peripheral region of the surface of the functional substrate on the substrate layer side, and the peripheral region of the surface of the functional substrate on the substrate layer side is The cell culture container according to claim 1, wherein no adhesive or pressure-sensitive adhesive is present on the enclosed area.   An opening is formed in a portion surrounded by a peripheral region of the surface of the container body member on the base material layer side of the functional substrate and a portion to which the surface is bonded via an adhesive or an adhesive. The cell culture container according to claim 2, wherein a region surrounded by the peripheral region of the surface on the base material layer side of the substrate is exposed through the opening.

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