JP5857642B2 - Cell culture container and manufacturing method thereof - Google Patents

Description

  The present invention relates to a method for producing a cell culture container.

Patent Document 1 discloses a cell culture support (a cell culture container such as a petri dish) coated with a layer of a functional polymer (temperature-responsive polymer) to be cultured on the surface.
By using this cell culture support, it can be easily detached from the cell support and recovered without destroying the cultured / proliferated cells simply by changing the temperature.
Patent Document 1 discloses a method in which cell culture containers are individually batch-processed to provide a functional organic compound layer.

JP-A-2-21865

As an efficient method for producing a cell culture vessel provided with a functional organic compound layer, a functional substrate such as a film or plate having a functional organic compound layer formed on a base material layer is produced in advance, There is a method of fixing to the bottom of the culture vessel. In FIG. 1, as an example of the prior art, a base layer 112 and a functional organic compound layer 111 are provided on the bottom of a dish-shaped cell culture vessel 100 having a bottom 101 and a peripheral side wall 102 and opened on the top. The case where the functional base 110 provided with is fixed is shown. At this time, the outer periphery of the functional base 110 needs to be slightly smaller than the inner periphery of the peripheral side wall 102. For this reason, it is difficult to form the functional organic compound layer 111 on the entire surface of the bottom 101 inside the container, and at least one portion of the bottom 101 on the surface inside the container that is in contact with the lower end of the peripheral side wall portion. The part is a part where the functional organic compound layer is not formed. As a result, there is a problem that desired functions such as cell adhesion and cell peelability are not sufficiently exhibited. For example, when cell culture is performed on a temperature-responsive polymer layer to form a cell sheet, and the cell sheet is peeled off and collected by temperature change, the cell sheet adheres to the bottom of the container where the temperature-responsive polymer layer does not exist. Therefore, there is a problem that it becomes difficult to separate and collect the cell sheet.
Therefore, the present invention provides a cell culture container having a functional organic compound layer provided on the entire bottom surface and a method for producing the same.

  The present inventors prepare a bottom portion having a functional organic compound layer fixed on the surface, a peripheral side wall portion at the periphery of the bottom portion, and at least a portion near the inner periphery of the lower end surface of the functional organic compound. It has been found that a cell culture vessel in which a functional organic compound layer is provided on the entire bottom surface can be obtained by providing the layer so as to be placed on the periphery of the layer, and the present invention has been completed.

The present invention includes the following inventions.
(1) a cell culture container,
A bottom provided with a functional organic compound layer extending along the direction of the bottom surface in the container;
A cell culture container provided with a peripheral side wall portion provided on a peripheral edge portion of the functional organic compound layer, at least a portion closer to the inner periphery of the lower end surface thereof standing on the peripheral edge of the bottom portion.

(2) The bottom is
A bottom plate member;
A function comprising a base material layer and a functional organic compound layer laminated on the surface of the base material layer, on the inner surface of the container of the bottom plate member, the function organic compound layer being disposed so as to face the inner side of the container A combination with a conductive substrate,
The cell culture container of (1).

(3) The cell culture container according to (1), wherein the bottom is a bottom plate member provided with a functional organic compound layer laminated on a surface inside the container.

(4) The cell culture container according to (1), wherein the bottom is a functional film including a film base layer and a functional organic compound layer laminated on the inner surface of the film base layer.

(5) It further includes a frame-like support portion having an upper end surface facing the lower end surface of the peripheral side wall portion,
The portion near the inner periphery of the lower end surface of the peripheral side wall portion is placed on the peripheral portion of the surface of the functional film on the functional organic compound layer side,
The peripheral part of the surface of the functional film on the film base layer side is placed on the part near the inner periphery of the upper end surface of the frame-shaped support part,
The peripheral edge of the functional film is gripped by the portion near the inner periphery of the lower end surface of the peripheral side wall portion and the portion near the inner periphery of the upper end surface of the frame-shaped support portion that faces the portion. (4) The cell culture container according to (4), wherein a portion near the outer periphery of the lower end surface of the peripheral side wall portion and a portion near the outer periphery of the upper end surface of the frame-shaped support portion facing the portion are joined.

(6) The portion in contact with the surface on the functional organic compound layer side of the functional film near the inner periphery of the lower end surface of the peripheral side wall includes an elastic material,
The portion close to the inner periphery of the upper end surface of the frame-shaped support portion and in contact with the surface on the film base layer side of the functional film contains an elastic material.
(5) The cell culture container.

(7) A method for producing a cell culture container,
A step of disposing a functional substrate including a base material layer and a functional organic compound layer laminated on the surface of the base material layer on the surface of the bottom plate member so that the functional organic compound layer faces the inside of the container; ,
Arranging the peripheral side wall portion so that at least a portion near the inner periphery of the lower end surface thereof is placed on the peripheral portion of the functional organic compound layer, and joining the bottom plate member and / or the functional substrate;
Including methods.

(8) A method for producing a cell culture container,
On the surface of the bottom plate member provided with the functional organic compound layer laminated on the inner surface of the container, the peripheral side wall portion, and at least the portion near the inner periphery of the lower end surface thereof is the peripheral portion of the functional organic compound layer. A step of placing the peripheral side wall portion and the bottom plate member,
Including methods.

(9) A method for producing a cell culture container,
A functional support film including a film base material layer and a functional organic compound layer laminated on the film base material layer, a peripheral side wall portion, and a frame-shaped support portion provided with an upper end surface facing the lower end surface of the peripheral side wall portion The process of preparing
A part of the functional film near the inner periphery of the lower end surface of the peripheral side wall portion is in contact with the peripheral portion of the surface on the functional organic compound layer side, and a frame-shaped support portion is formed on the peripheral portion of the surface on the film base material layer side. The portion near the inner periphery of the upper end surface of the peripheral wall portion is in contact with the portion near the outer periphery of the lower end surface of the peripheral side wall portion and the portion near the outer periphery of the upper end surface of the frame-shaped support portion at a position outside the outer periphery of the functional film. A step of arranging to
While holding the peripheral edge of the functional film by the inner peripheral portion of the lower end surface of the peripheral side wall portion and the inner peripheral portion of the upper end surface of the frame-shaped support portion, Joining the portion and the portion near the outer periphery of the upper end surface of the frame-like support portion.

(10) The portion in contact with the surface on the functional organic compound layer side of the functional film near the inner periphery of the lower end surface of the peripheral side wall includes an elastic material,
The portion close to the inner periphery of the upper end surface of the frame-shaped support portion and in contact with the surface on the film base layer side of the functional film contains an elastic material.
Method (9).

  According to the present invention, there is provided a cell culture vessel in which a functional organic compound layer is provided on the entire bottom surface of the vessel.

FIG. 1 is a schematic cross-sectional view showing an example of a conventional technique for fixing a functional substrate to the bottom surface of a dish-shaped cell culture container (petri dish) opened on the top surface. It is a figure which shows an example (dish shape) of the whole shape of the cell culture container of this invention. It is a figure which shows an example (flask shape) of the whole shape of the cell culture container of this invention. An example of the manufacturing method of the cell culture container which concerns on 1st Embodiment of this invention which makes a bottom part the combination of a baseplate member and a functional base | substrate provided with a base material layer and a functional organic compound layer is shown. The other example of the manufacturing method of the cell culture container which concerns on 1st Embodiment of this invention which makes a bottom part the combination of a baseplate member and a functional base | substrate provided with a base material layer and a functional organic compound layer is shown. The other example of the manufacturing method of the cell culture container which concerns on 1st Embodiment of this invention which makes a bottom part the combination of a baseplate member and a functional base | substrate provided with a base material layer and a functional organic compound layer is shown. An example of the manufacturing method of the cell culture container which concerns on 2nd Embodiment of this invention which uses a baseplate member provided with the functional organic compound layer laminated | stacked on the surface inside a container as a bottom part is shown. The other example of the manufacturing method of the cell culture container which concerns on 2nd Embodiment of this invention which uses a baseplate member provided with the functional organic compound layer laminated | stacked on the surface inside a container as a bottom part is shown. An example of the manufacturing method of the cell culture container which concerns on 3rd Embodiment of this invention which uses a functional film provided with a film base material layer and a functional organic compound layer as a bottom part is shown. The other example of the manufacturing method of the cell culture container which concerns on 3rd Embodiment of this invention which uses a functional film provided with a film base material layer and a functional organic compound layer as a bottom part is shown. The application example to the flask type container of the manufacturing method of the cell culture container which concerns on 3rd Embodiment of this invention which makes a bottom the functional film provided with a film base material layer and a functional organic compound layer is shown. The application example to the flask type container of the manufacturing method of the cell culture container which concerns on 3rd Embodiment of this invention which makes a bottom the functional film provided with a film base material layer and a functional organic compound layer is shown.

  In the following description, the features of the present invention will be described with reference to the drawings as needed. However, each drawing is a schematic view, and the size and shape of each portion are appropriately exaggerated for easy understanding. It shows.

<Shape of cell culture vessel>
The overall shape of the cell culture container of the present invention will be described.
As long as the cell culture container manufactured by this invention is provided with a bottom part and a surrounding side wall part, the whole shape is not specifically limited.

  As shown in FIG. 2, an example of the cell culture container of the present invention is a dish-shaped cell culture container 200 that includes a bottom part 201 and a peripheral side wall part 202 and is opened in the upper end direction of the peripheral side wall part 202. A functional organic compound layer 210 is provided on the inner bottom surface of the bottom 201.

  As shown in FIG. 3, another example of the cell culture container of the present invention is joined to the bottom 301, the peripheral side wall 302 standing on the periphery of the bottom 301, and the upper end of the peripheral side wall 302. A “flask type” having a top surface portion 303 disposed opposite to the bottom portion 301, a through hole 304 formed in a part of the peripheral side wall portion 302, and a neck portion 305 extending from the periphery of the through hole 304 to the outside of the container portion. Is a flask-type cell culture container 320 including a container portion 300 having a shape called “a” and a lid 310 that is detachably attached via a locking portion 306 formed on the neck portion 305. 3B is a cross-sectional view taken along the line I-I ′ of the container 300, and FIG. 3C is a cross-sectional view taken along the line II-II ′. Although not shown, a functional organic compound layer 330 is provided on the bottom surface of the bottom 301 inside the container.

  In the present invention, the “peripheral side wall” may consist of only the peripheral side wall, or may be a peripheral side wall provided with a through-hole in a part and provided with a neck, The peripheral wall part of the shape where the upper end was closed by the top | upper surface part etc. may be sufficient.

  In the following description, an embodiment of the simplest structure, which is an embodiment of forming a functional organic compound layer on the inner bottom surface of the dish-shaped cell culture container 200, will be mainly described. Is not limited.

<Material of cell culture container>
The material for forming other members of the cell culture container such as the bottom plate member, the peripheral side wall portion, the frame-like support portion, 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.

<Functional organic compound layer>
In the present invention, the functional organic compound layer is provided on the inner surface of the container of the bottom plate member or the surface of the functional substrate.

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 forms a desired stimulus-responsive polymer by polymerization and an organic solvent that can dissolve the monomer, and this is applied to the bottom plate member according to a conventional coating method. Alternatively, it is applied to the surface of the base material layer to form a coating film, and then the monomer in the coating film is polymerized by an appropriate means such as irradiation with the coating film to form a polymer. It can be formed by causing a grafting reaction between the surface of the material layer and the polymer.

<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-made bottom plate member or substrate layer, it is physically adsorbed on the surface of the bottom plate member or substrate layer beforehand. A primer layer containing a polysiloxane containing a functional group in the side chain that is capable of reacting with a hydroxyl group at the end of the ethylene glycol chain to form a covalent bond 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.

<Functional substrate>
In 1st Embodiment of this invention, a functional base | substrate provided with a base material layer and a functional organic compound layer is used.
In the present invention, the “base” may be a film or a plate 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 film is suitably used in the third embodiment of the present invention.

  The functional substrate can be produced by forming a functional organic compound layer on a resin base layer by an appropriate method. Between the resin base material layer and the functional organic compound layer, one or more other layers (for example, the primer layer for fixing the hydrophilic compound layer to the resin base material layer) as necessary. May be present.

  The planar shape of the functional substrate can be any shape depending on the shape of the region 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.

  The substrate layer is appropriately selected according to the final functional substrate. If the functional substrate is plate-like, a plate-like resinous substrate layer is used, and if the functional substrate is a film-like, a film-like resinous substrate layer (hereinafter referred to as “film substrate layer”) is used. It is done.

  The base material layer should just contain the resin 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 the like.

  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 that imparts flexibility, for example, 5 to 500 μm, more preferably 20 to 500 μm, and particularly preferably 50 to 250 μm.

The function and structure of the functional organic compound layer provided in the functional substrate are as described above.
Formation of the functional organic compound layer on the surface of the base material layer can be performed by an appropriate method such as the method described in detail for the stimulus-responsive polymer layer and the hydrophilic compound layer. Between the base material layer and the functional organic compound layer, a layer necessary for laminating the functional organic compound layer such as a primer layer may be interposed.

<Bottom plate member on which functional organic compound layer is laminated>
In 2nd Embodiment of this invention, the baseplate member in which the functional organic compound layer was formed in the surface inside a container is used.

  In this case, the bottom plate member only needs to include a resin material capable of forming the above-described functional organic compound layer on one surface, and is formed of the resin material among the materials described for the material of the cell culture container. It is particularly preferable.

The function and structure of the functional organic compound layer on the bottom plate member are as described above.
Formation of the functional organic compound layer on the surface of the bottom plate member can be performed by an appropriate method such as the method described in detail regarding the stimulus-responsive polymer layer and the hydrophilic compound layer. Between the surface of the bottom plate member and the functional organic compound layer, a layer necessary for stacking the functional organic compound layer such as a primer layer may be interposed.

<Characteristics of the cell culture container of the present invention>
The cell culture container of the present invention comprises:
A bottom provided with a functional organic compound layer extending along the direction of the bottom surface in the container;
At least a portion near the inner periphery of the lower end surface provided on the periphery of the bottom portion includes a peripheral side wall portion mounted on the periphery of the functional organic compound layer.

  A cell culture container having this feature has a functional organic compound layer without a gap not only on the entire bottom surface of the container, that is, not only on the central portion of the bottom surface of the container but also on the portion contacting the lower end of the peripheral side wall. Unlike the prior art as shown in FIG. 1 in which it was difficult to provide a functional organic compound layer up to the lower end of the peripheral side wall portion on the bottom surface of the container, desired cell adhesion, cell peelability, etc. It is possible to fully exhibit the functions.

The cell culture container of the present invention is roughly divided into three embodiments according to the form of the bottom.
In 1st Embodiment of the cell culture container of this invention, a bottom part is the combination of a baseplate member and a functional base | substrate.
In 2nd Embodiment of the cell culture container of this invention, a bottom part is a baseplate member provided with the functional organic compound layer laminated | stacked on the surface inside a container.
In 3rd Embodiment of the cell culture container of this invention, a bottom part is a functional film.
Hereinafter, the structure and manufacturing method of each embodiment will be specifically described.

<First Embodiment>
A first embodiment of the present invention will be described with reference to FIGS.
Cell culture containers 420, 520, and 620 manufactured in the examples shown in the sectional views of FIGS. 4 to 6 are dish-shaped cell culture containers 200 as shown in FIG.

In the embodiment of FIG. 4, the bottom plate member 401 and the functional substrate 402 are combined to form the bottom of the cell culture container.
The functional substrate 402 includes at least a base material layer 412 and a functional organic compound layer 411.

  The cell culture container 420 has a step (FIGS. 4A and 4B) in which the functional substrate 402 is arranged and fixed on the inner surface of the bottom plate member 401, and the peripheral side wall 403, preferably at its lower end surface 430. It arrange | positions so that it may mount on the peripheral part of the functional organic compound layer 411 over the perimeter, and it joins to the functional base | substrate 402 through the functional organic compound layer 411.

  In the form shown in FIG. 4, the bottom plate member 401 and the functional substrate 402 are fixed, and the peripheral edge portion of the functional organic compound layer 411 of the functional substrate 402 and the lower end surface 430 of the peripheral side wall portion 403 are fixed.

  The method for fixing the bottom plate member 401 and the functional substrate 402 is not particularly limited. For example, a method of fixing both of them via an adhesive, a pressure-sensitive adhesive, a heat sealable resin, or the like, or a resin base plate member 401 and a resin base material layer 412 are made of a resin such as ultrasonic welding or laser welding. The method of joining using joining technology is mentioned. Although not shown in the figure, when fixing members through adhesives, pressure-sensitive adhesives, heat-sealable resins, etc., there are adhesive layers, pressure-sensitive adhesive layers, heat-sealable resin layers, etc. between the fixed members. Appropriately exists.

  The method for joining the peripheral edge portion of the functional organic compound layer 411 of the functional substrate 402 and the lower end surface 430 of the peripheral side wall portion 403 is also not particularly limited. For example, a method of fixing both via an adhesive, a pressure-sensitive adhesive, a heat-sealable resin, or the like, or a resin-made base material layer 412 and a resin-made peripheral side wall 403 via a functional organic compound layer 411 And a method of bonding using a resin bonding technique such as ultrasonic welding or laser welding.

  In the example of FIG. 4, the entire lower end surface 430 of the peripheral side wall portion 403 is in contact with the peripheral edge portion of the functional organic compound layer 411, but is not limited thereto. It is only necessary that the peripheral portion is in contact with the peripheral edge of the functional organic compound layer 411.

  5 and 6, only the portion near the inner periphery of the lower end surface of the peripheral side wall portion is placed on the peripheral portion of the functional organic compound layer, and the portion near the outer periphery of the lower end surface of the peripheral side wall portion is joined to the bottom plate member. An example is shown.

  In the example shown in FIG. 5, the bottom plate member 401, the functional base 502, and the peripheral side wall portion 403 are overlapped in the order of FIG. The outer periphery of 430 is located outside the outer periphery of the functional substrate 502 including the functional organic compound layer 511 and the base material layer 512, and the inner periphery of the lower end surface 430 of the peripheral side wall 403 is functional. It is located inside the outer periphery of the substrate 502.

  In the example shown in FIG. 5, first, the bottom plate member 401 and the functional base 502 are arranged such that the periphery of the bottom plate member 401 is located outside the periphery of the functional base 502 (FIG. 5B). Subsequently, the peripheral side wall 403 has a portion near the inner periphery of the lower end surface 430 placed on the peripheral edge of the functional organic compound layer 511 of the functional substrate 502, and a portion near the outer periphery of the lower end surface 430 is the functional substrate 502. It arrange | positions so that it may be located outside the periphery of FIG. At this time, the portion near the outer periphery of the lower end surface 430 of the peripheral side wall portion 403 faces the peripheral portion of the container inner surface of the bottom plate member 401 (FIG. 5D). The part near the outer periphery of the lower end surface 430 of the peripheral side wall part 403 and the peripheral part of the inner surface of the container of the bottom plate member 401 are joined by an adhesive, an adhesive, a heat sealable resin, or the like, or the peripheral side wall part 403 and the bottom plate When the members 401 are both made of resin, the cell culture vessel 520 is completed by joining by means of ultrasonic welding, laser welding or the like (FIG. 5E).

  FIG. 6 shows a modification of the example of FIG. 5 in which the bottom plate member and the portion near the outer periphery of the lower end surface of the peripheral side wall portion are joined. In the example of FIG. 6, the height position of the inner peripheral portion 631 on the lower end surface of the peripheral side wall portion 603 and the outer peripheral portion 632 are different, and the latter 632 protrudes in the bottom direction relative to the former 631. The latter 632 is formed so as to come into contact with the peripheral portion of the inner surface of the container of the bottom plate member 401 with 631 placed on the peripheral portion of the functional organic compound layer 511 of the functional substrate 502.

  In the example shown in FIG. 6, first, the bottom plate member 401 and the functional base 502 are arranged so that the periphery of the bottom plate member 401 is located outside the periphery of the functional base 502 (FIG. 6B). Subsequently, the peripheral wall portion 603 has a portion 631 near the inner periphery of the lower end surface thereof placed on the peripheral edge portion of the functional organic compound layer 511 of the functional substrate 502, and a portion 632 near the outer periphery of the lower end surface is formed of the bottom plate member 401. It arrange | positions so that the peripheral part of a container inner surface may be contact | abutted (FIG. 6D). Next, the portion 632 near the outer periphery of the lower end surface of the peripheral side wall portion 603 and the peripheral portion on the inner surface of the container of the bottom plate member 401 are joined with an adhesive, an adhesive, a heat sealable resin, or the like, or the peripheral side wall portion 603 and When both the bottom plate members 401 are made of resin, the cell culture container 620 is completed by joining by means of ultrasonic welding, laser welding, or the like (FIG. 6E).

  In the example shown in FIGS. 5 and 6, the peripheral edge of the functional substrate 502 is gripped by the inner peripheral portion of the lower end surface of the peripheral side wall portion and the surface of the bottom plate member 401. Therefore, it is not essential that the bottom plate member 401 and the functional substrate 502 are bonded to each other.

Second Embodiment
A second embodiment of the present invention will be described with reference to FIGS.
Cell culture containers 720 and 820 manufactured in the example shown in the cross section in FIGS. 7 to 8 are dish-shaped cell culture containers 200 as shown in FIG.

In the embodiment of FIG. 7, a bottom plate member 701 including a functional organic compound layer 711 stacked on the inner surface of the container is prepared (FIG. 7B). The functional organic compound layer 711 is formed on the entire surface of the bottom plate member 701 inside the container. The peripheral side wall part 403 is arranged so that the lower end surface 430 thereof is placed on the peripheral edge part of the functional organic compound layer 711, and the peripheral side wall part 403 and the bottom plate member 701 are arranged via the functional organic compound layer 711 in the first embodiment. The cell culture container 720 is completed by the same means as described for the form (FIG. 7D).
The function and structure of the functional organic compound layer 711 are as described above.

  In the example of FIG. 7, the entire lower end surface 430 of the peripheral side wall portion 403 is bonded in a state of being in contact with the peripheral edge portion of the functional organic compound layer 711, but is not limited thereto. It suffices that at least the portion near the inner periphery of the end surface 430 is in contact with the peripheral portion of the functional organic compound layer 711.

  FIG. 8 shows an example in which the portion near the inner periphery of the lower end surface of the peripheral side wall portion is placed on the peripheral portion of the functional organic compound layer, and the portion near the outer periphery of the lower end surface of the peripheral side wall portion is joined to the bottom plate member. . In the example shown in FIG. 8, the outer periphery of the bottom plate member 701 and the outer periphery of the lower end surface 430 of the peripheral side wall part 403 are in a plan view when the bottom plate member 701 and the peripheral side wall part 403 are overlapped as shown in FIG. 8D. , Respectively, are located outside the outer periphery of the functional organic compound layer 811. And the inner periphery of the lower end surface 430 of the peripheral side wall part 403 is located inside the outer periphery of the functional organic compound layer 811. In this embodiment, first, the functional organic compound layer 811 is formed on the inner surface of the bottom plate member 701 such that the periphery of the bottom plate member 701 is located outside the periphery of the functional organic compound layer 811 (FIG. 8B). Subsequently, the peripheral wall portion 403 has a portion near the inner periphery of the lower end surface 430 placed on the peripheral edge of the functional organic compound layer 811, and a portion near the outer periphery of the lower end surface 430 is more than the peripheral edge of the functional organic compound layer 811. Are also positioned outside (FIG. 8D). At this time, the part near the outer periphery of the lower end surface 430 of the peripheral side wall part 403 faces the peripheral part of the inner surface of the bottom plate member 701 (FIG. 8D). By joining the portion near the outer periphery of the lower end surface 430 of the peripheral wall portion 403 and the peripheral portion of the inner surface of the container of the bottom plate member 701 by the same means as described in the first embodiment, the cell culture container 820 is joined. Complete (FIG. 8E).

<Third Embodiment>
A third embodiment of the present invention will be described with reference to FIGS.
Cell culture containers 920 and 1020 manufactured in the example shown in the cross section in FIGS. 9 to 10 are dish-shaped cell culture containers 200 as shown in FIG.

  In 3rd Embodiment, a functional film provided with the functional organic compound layer laminated | stacked on the surface inside a container of a film base material layer and this film base material layer is used as a bottom part of a cell culture container. And The characteristics of the functional film are as described above.

  When the functional film is used as the bottom of the cell culture container, it is preferable to fix the functional film by the method shown in FIG. 9 so that the functional film does not fall off. First, the functional film 901 including the film base material layer 912 and the functional organic compound layer 911 laminated on the film base material layer, the peripheral side wall portion 403, and the lower end surface 430 of the peripheral side wall portion 403 are opposed to each other. A frame-like support portion 940 having an upper end surface 930 is prepared (FIG. 9A). Here, in the plan view when superimposed as shown in FIG. 9B, the outer periphery of the functional film 901 is outside the inner periphery of the lower end surface 430 of the peripheral side wall portion 403 and the upper end surface 930 of the frame-shaped support portion 940. Yes, it is configured to be inside the outer periphery. In the present invention, the “frame-shaped support portion” has a frame shape having an upper end surface facing the lower end surface of the peripheral side wall portion.

  The functional film 901, the peripheral side wall part 403, and the frame-shaped support part 940 are arranged on the peripheral part of the surface of the functional film 901 on the functional organic compound layer 911 side, and the inner periphery of the lower end surface 430 of the peripheral side wall part 403. A portion closer to the inner periphery of the upper end surface 930 of the frame-shaped support portion 940 comes into contact with a peripheral portion of the surface on the film base material layer 912 side, and closer to the outer periphery of the lower end surface 430 of the peripheral side wall portion 403. And the portion near the outer periphery of the upper end surface 930 of the frame-shaped support portion 940 are arranged so as to face each other at a position outside the outer periphery of the functional film 901 (FIG. 9B).

  Subsequently, while the peripheral edge of the functional film 901 is gripped by the inner peripheral portion of the lower end surface 430 of the peripheral side wall portion 403 and the inner peripheral portion of the upper end surface 930 of the frame-shaped support portion 940, The portion near the outer periphery of the lower end surface 430 of the portion 403 and the portion near the outer periphery of the upper end surface 930 of the frame-shaped support portion 940 are joined by the same means as in the first embodiment, thereby completing the cell culture container 920. (FIG. 9C). The functional film 901 is preferably configured such that the entire periphery is gripped by the portion near the inner periphery of the lower end surface 430 of the peripheral side wall portion 403 and the portion near the inner periphery of the upper end surface 930 of the frame-shaped support portion 940. Is prevented from falling off.

  Although not shown, in order to facilitate the joining of the portion near the outer periphery of the lower end surface 430 of the peripheral side wall portion 403 and the portion near the outer periphery of the upper end surface 930 of the frame-shaped support portion 940, as shown in FIG. The configuration in which the portion near the outer periphery of the lower end surface 430 of the peripheral side wall portion 403 protrudes in the bottom direction from the portion near the inner periphery, and / or the portion near the outer periphery of the upper end surface 930 of the frame-shaped support portion 940 By adopting a configuration that protrudes in the direction of the peripheral side wall portion from the portion near the inner periphery, the portion near the outer periphery of the lower end surface 430 of the peripheral side wall portion 403 and the outer periphery of the upper end surface 930 of the frame-shaped support portion 940 are arranged. You may make it contact | abut with a part.

  Furthermore, the modification for strengthening the holding | grip of the functional film peripheral part by the lower end surface of a surrounding side wall part and the upper end surface of a frame-shaped support part is shown in FIG. In this modification, the portion near the inner periphery of the lower end surface 1030 of the peripheral side wall portion 1003 preferably includes the elastic material 1031 that is in close contact with the surface on the functional organic compound layer 911 side of the functional film 901. In addition, the portion of the upper end surface 1040 of the frame-shaped support portion 1004 that is closer to the inner periphery preferably includes the elastic material 1041 that is in close contact with the surface on the film base layer 912 side over the entire periphery. Examples of the elastic materials 1031 and 1041 include silicone rubber materials such as polydimethylsiloxane. The close contact with the elastic materials 1031 and 1041 makes it possible to more firmly hold the peripheral edge of the functional film and prevent liquid leakage from the peripheral edge of the functional film.

<Flask type cell culture vessel>
Although FIGS. 4-10 shows the example which manufactures a dish-shaped cell culture container, it is not limited to this. It is obvious to those skilled in the art that other cell culture containers such as flask-type cell culture containers can be produced in the same manner. For example, by the method shown in FIGS. 4 to 10, the bottom 301 having a functional organic compound layer extending along the direction of the bottom surface inside the container, the through-hole 104 and the neck 105 are opened upward. The flask-shaped container part 300 can be completed by combining the peripheral wall part 302 thus formed, and subsequently joining the top surface part 303 to the open end above the peripheral wall part 302 and closing the open end.

  Alternatively, by the method shown in FIGS. 4 to 10, the bottom portion 301 having a functional organic compound layer that extends along the direction of the bottom surface inside the container, and the upper portion with the through-hole 104 and the neck portion 105, is the top. The flask-type container 300 can be completed by combining the peripheral wall 302 closed with the surface 303. For example, in 3rd Embodiment of this invention which fixes a functional film to the lower end surface of a surrounding side wall part using a frame-shaped support part, as shown in FIG.11 and FIG.12, by the top | upper surface part provided with the through-hole and the neck part. The closed peripheral side wall part 1101, the functional film 1102, and the frame-like support part 1103 having the upper end face facing the lower end face of the peripheral side wall part 1101 are arranged, and the peripheral part of the functional film 1102 is connected to the peripheral side wall. A portion near the outer periphery of the lower end surface 1230 of the peripheral side wall portion 1101 and a frame shape while being gripped by a portion near the inner periphery of the lower end surface 1230 of the portion 1101 and a portion near the inner periphery of the upper end surface 1240 of the frame-shaped support portion 1103 The flask-shaped container portion 1220 can be manufactured by joining the portion near the outer periphery of the upper end surface 1240 of the support portion 1103 by the above-described means.

200, 320 ... cell culture vessel 201, 301 ... bottom 202, 302, 403, 603, 1003, 1101 ... peripheral side wall 401, 701 ... bottom plate member 402, 502 ... functional substrate 901, 1102 ... functional films 940, 1004, 1103 ... frame-like support portions 210, 330, 411, 511, 711, 811, 911 ... functional organic compound layers

Claims (4)

A cell culture container,
Functionality provided with a functional organic compound layer extending along the direction of the bottom surface in the container , and a film base material layer and a functional organic compound layer laminated on the inner surface of the film base material layer A bottom that is a film ,
A peripheral side wall portion that is erected on the peripheral edge of the bottom portion and at least a portion near the inner periphery of the lower end surface thereof is placed on the peripheral edge portion of the functional organic compound layer ;
A frame-like support portion having an upper end surface facing the lower end surface of the peripheral side wall portion;
Equipped with a,
The portion near the inner periphery of the lower end surface of the peripheral side wall portion is placed on the peripheral portion of the surface of the functional film on the functional organic compound layer side,
The peripheral part of the surface of the functional film on the film base layer side is placed on the part near the inner periphery of the upper end surface of the frame-shaped support part,
The peripheral edge of the functional film is gripped by the portion near the inner periphery of the lower end surface of the peripheral side wall portion and the portion near the inner periphery of the upper end surface of the frame-shaped support portion that faces the portion. A portion near the outer periphery of the lower end surface of the peripheral side wall portion and a portion near the outer periphery of the upper end surface of the frame-shaped support portion facing the portion are joined,
The portion close to the inner periphery of the lower end surface of the peripheral side wall portion and in contact with the surface on the functional organic compound layer side of the functional film contains an elastic material,
The portion close to the inner periphery of the upper end surface of the frame-shaped support portion and in contact with the surface on the film base layer side of the functional film contains an elastic material.
Cell culture container. The portion in contact with the surface of the functional film on the side of the functional organic compound layer near the inner periphery of the lower end surface of the peripheral side wall portion includes an elastic material over the entire circumference.
The portion in contact with the surface on the film base layer side of the functional film near the inner periphery of the upper end surface of the frame-shaped support portion includes an elastic material over the entire circumference.
The cell culture container according to claim 1. A method for producing a cell culture container, comprising:
A functional support film including a film base material layer and a functional organic compound layer laminated on the film base material layer, a peripheral side wall portion, and a frame-shaped support portion provided with an upper end surface facing the lower end surface of the peripheral side wall portion The process of preparing
A part of the functional film near the inner periphery of the lower end surface of the peripheral side wall portion is in contact with the peripheral portion of the surface on the functional organic compound layer side, and a frame-shaped support portion is formed on the peripheral portion of the surface on the film base material layer side. The portion near the inner periphery of the upper end surface of the peripheral wall portion is in contact with the portion near the outer periphery of the lower end surface of the peripheral side wall portion and the portion near the outer periphery of the upper end surface of the frame-shaped support portion at a position outside the outer periphery of the functional film. A step of arranging to
While holding the peripheral edge of the functional film by the inner peripheral portion of the lower end surface of the peripheral side wall portion and the inner peripheral portion of the upper end surface of the frame-shaped support portion, and a step of bonding the part and the frame-shaped outer peripheral side of the portion of the upper surface of the support portion seen including,
The portion close to the inner periphery of the lower end surface of the peripheral side wall portion and in contact with the surface on the functional organic compound layer side of the functional film contains an elastic material,
The portion close to the inner periphery of the upper end surface of the frame-shaped support portion and in contact with the surface on the film base layer side of the functional film contains an elastic material.
Method. The portion in contact with the surface of the functional film on the side of the functional organic compound layer near the inner periphery of the lower end surface of the peripheral side wall portion includes an elastic material over the entire circumference.
The portion in contact with the surface on the film base layer side of the functional film near the inner periphery of the upper end surface of the frame-shaped support portion includes an elastic material over the entire circumference.
The method of claim 3.

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