JP2021023287A - Cell culture scaffolding material and cell culture container - Google Patents

Abstract

To provide cell culture scaffolding materials that have excellent fixation of cells after seeding and can increase a cell proliferation rate.SOLUTION: Provided is a cell culture scaffolding material containing synthetic resin, having a zeta potential at pH 7 of -60 mV or higher, and having a plurality of pores with an average diameter of 10 nm or more and 5 μm or less.SELECTED DRAWING: None

Description

The present invention relates to a cell culture scaffold material used for culturing cells. The present invention also relates to a cell culture container using the above-mentioned cell culture scaffold material.

In recent years, cell medicine and next-generation medicine using stem cells have been attracting attention. Among them, human pluripotent stem cells (hPSC) such as regenerative medicine human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC), or differentiated cells derived from them, are used for drug discovery and regenerative medicine. Is expected to be applied. In order to achieve such an application, it is necessary to culture and proliferate pluripotent stem cells and differentiated cells safely and with good reproducibility.

In particular, for industrial use of regenerative medicine, it is necessary to handle a large amount of stem cells, so it is necessary to support the proliferation of pluripotent stem cells using natural polymer materials, synthetic polymer materials, or feeder cells. It becomes. Therefore, various culture methods using scaffolding materials composed of natural polymer materials, synthetic polymer materials, and the like have been studied.

For example, Patent Document 1 below describes a molded product made of a polyvinyl acetal compound or a molded product made of the polyvinyl acetal compound and a water-soluble polysaccharide, and the degree of acetalization of the polyvinyl acetal compound is 20 to 60 mol%. A carrier for cell culture is disclosed.

Patent Document 2 below discloses a composition containing the first fibrous polymer scaffolding material, wherein the fibers of the first fibrous polymer scaffolding material are aligned. It is described that the fibers constituting the first fibrous polymer scaffold material are composed of an aliphatic polyester such as polyglycolic acid or polylactic acid.

Further, Patent Document 3 below describes a cell culture method for maintaining the undifferentiated state of pluripotent stem cells, which is pluripotent on an incubator having a surface coated with a polyrotaxan block copolymer. A method including the step of culturing stem cells is disclosed.

Japanese Unexamined Patent Publication No. 2006-314285 Special Table 2009-524507 Japanese Unexamined Patent Publication No. 2017-23008

By the way, when the scaffold material is made of a natural polymer material, the colonization of cells after seeding can be enhanced. In particular, it is known that when an adhesive protein such as laminin or vitronectin or a mouse sarcoma-derived matrigel is used as a natural polymer material, the cell colonization after seeding is extremely high. On the other hand, natural polymer materials are expensive, have large variations between lots because they are naturally derived substances, and have safety concerns due to animal-derived components.

On the other hand, scaffolding materials using synthetic resins have better operability, lower cost, less variation between lots, and excellent safety than scaffolding materials using natural polymer materials. .. However, in scaffolding materials using synthetic resins as in Patent Documents 1 to 3, the synthetic resin swells excessively because the synthetic resin having high hydrophilicity is used. Further, since the synthetic resin has a lower affinity with cells than the natural polymer material, the cell mass may be exfoliated during culturing. As described above, the scaffold material using the synthetic resin has low colonization after seeding of the cells, and the cells may not proliferate sufficiently.

An object of the present invention is to provide a scaffold material for cell culture and a container for cell culture, which are excellent in colonization after seeding of cells and can increase the proliferation rate of cells.

The cell culture scaffold material according to the present invention contains a synthetic resin and has a plurality of pores having a zeta potential at pH 7 of −60 mV or more and an average diameter of 10 nm or more and 5 μm or less.

In certain aspects of the cell culture scaffold material according to the present invention, the zeta potential at pH 7 is +30 mV or less.

In another specific aspect of the cell culture scaffold material according to the present invention, the elastic modulus at 25 ° C. measured by the nanoindentation method is 0.01 GPa or more.

In still another specific aspect of the cell culture scaffold material according to the present invention, the synthetic resin contains Bronsted basic groups in a constituent unit of 0.2 mol% or more and 20 mol% or less.

In still another specific aspect of the cell culture scaffold material according to the present invention, the water swelling rate is 100% or less.

Yet another particular aspect of the cell culture scaffold material according to the present invention is substantially free of animal-derived material.

In yet another particular aspect of the cell culture scaffold material according to the present invention, the synthetic resin comprises a vinyl polymer.

In yet another particular aspect of the cell culture scaffold material according to the present invention, the synthetic resin comprises at least a polyvinyl alcohol derivative or a poly (meth) acrylic resin.

In yet another particular aspect of the cell culture scaffold material according to the present invention, the synthetic resin comprises a polyvinyl acetal resin.

In still another specific aspect of the cell culture scaffold material according to the present invention, the number of the pores per unit area is 0.01 / μm ² or more and 1000 / μm ² or less.

The cell culture container according to the present invention includes a cell culture scaffold material configured according to the present invention in at least a part of the cell culture area.

According to the present invention, it is possible to provide a scaffold material for cell culture and a container for cell culture, which are excellent in fixability after seeding of cells and can increase the proliferation rate of cells.

It is a schematic front sectional view which shows the container for cell culture which concerns on one Embodiment of this invention. 6 is an SEM observation image of the cell culture scaffold material obtained in Example 1 at a magnification of 1000 times. 6 is an SEM observation image of the scaffold material for cell culture obtained in Example 1 at a magnification of 10000 times.

Hereinafter, the present invention will be clarified by explaining specific embodiments of the present invention with reference to the drawings.

The scaffold material for cell culture according to the present invention is a scaffold material (scaffold) containing a synthetic resin. In the above-mentioned scaffold material for cell culture, the zeta potential at pH 7 is −60 mV or more. Further, the cell culture scaffold material has a plurality of pores having an average diameter of 10 nm or more and 5 μm or less.

Since the scaffold material for cell culture of the present invention has the above-mentioned structure, it has excellent colonization after seeding of cells and can increase the proliferation rate of cells.

Scaffolding materials for cell culture using conventional natural polymer materials can improve the colonization of cells after seeding, but they are expensive, and because they are naturally derived substances, there is a large variation between lots, and animals. There are safety concerns due to the ingredients of origin. On the other hand, in a scaffold material using a synthetic resin, the synthetic resin swells excessively or has a low affinity with cells, so that the cell mass may exfoliate during culturing. Therefore, the scaffolding material using the synthetic resin has low colonization after seeding of the cells, and the cells may not proliferate sufficiently.

On the other hand, the cell culture scaffold material according to the present invention has a zeta potential at pH 7 of −60 mV or higher.

Furthermore, the present inventors have focused on the structure of such a scaffold material for cell culture, and have found that the cell proliferation rate can be increased by having a plurality of pores having an average diameter of 10 nm or more and 5 μm or less. I found it.

The reason for this is not clear, but by adopting a structure in which a plurality of pores having an average diameter in a specific range are provided as described above, the cell adhesion portion, which is a portion in which the pores are not provided, can be used. , Both can be formed with the non-adherent portion of the cell, which is the portion where the pores are provided. Therefore, the cells can move freely while maintaining the cell colonization at the adhesion portion of the cells. Therefore, it is considered that the migration ability of cells can be enhanced and the proliferation ability of cells can be enhanced.

Therefore, according to the scaffold material for cell culture of the present invention, the adhesiveness to the cells after seeding can be enhanced, and the proliferation rate of the cells can be enhanced.

Further, in the present invention, since the synthetic resin can be used as described above, the operability is good, the cost is low, and the variation between lots is small as compared with the scaffolding material using the natural polymer material. It has excellent safety.

It is preferable that the scaffold material for cell culture of the present invention is substantially free of animal-derived raw materials. In that case, the scaffold material for cell culture can be made even more excellent in safety. In addition, "substantially free of animal-derived raw materials" means that the animal-derived raw materials in the cell culture scaffold material are 3% by weight or less. However, it is more preferable that the scaffold material for cell culture of the present invention does not contain any animal-derived raw materials.

In the scaffold material for cell culture of the present invention, the average diameter of the pores is preferably 50 nm or more, more preferably 200 nm or more, still more preferably 500 nm or more, preferably 4.5 μm or less, more preferably 4 μm or less, still more preferably. It is 3.5 μm or less. When the average diameter of the pores is within the above range, the colonization after seeding of the cells can be further enhanced, and the proliferation rate of the cells can be further enhanced.

Further, in the cell culture scaffold material of the present invention, the number of pores per unit area is preferably 0.01 / μm ² or more, more preferably 0.1 / μm ² or more, and further preferably 1. / Μm ² or more, particularly preferably 10 pieces / μm ² or more, preferably 1000 pieces / μm ² or less, more preferably 500 pieces / μm ² or less, still more preferably 100 pieces / μm ² or less. When the number of pores per unit area is within the above range, the colonization property after seeding of the cells can be further enhanced, and the proliferation rate of the cells can be further enhanced.

In the present specification, the average diameter and number of pores can be measured by a scanning electron microscope (SEM), a transmission electron microscope (TEM), or the like. In this case, for example, the average value of the pore diameters of any 100 pores confirmed by microscopic observation with a scanning electron microscope (SEM) or a transmission electron microscope (TEM) is defined as the average diameter of the pores. Can be done. The number of pores per unit area is obtained by dividing the number of pores in the observation region (5 μm × 5 μm) by the area of the observation region. The number of pores can be, for example, an average value obtained by repeating the same operation 10 times. When measuring the average diameter and number of pores, pores having a pore diameter of 10 nm or more and 5 μm or less shall be judged as pores.

Further, the plurality of pores can be formed, for example, by applying a solution containing a synthetic resin and alcohol on a substrate, freezing it with liquid nitrogen or the like, and then freeze-drying it. In this case, it is considered that the portion where the alcohol has escaped forms pores. When a synthetic resin that does not easily expand, such as a polyvinyl acetal resin, is used, the size of the pores can be made more constant, so that the pores having an average diameter controlled can be formed more easily. Can be done.

The average diameter and number of pores can be controlled by adjusting the type and amount of alcohol added, adding a small amount of water to the alcohol, and the like.

Further, in the present invention, a scaffold material for cell culture may be formed by crushing a synthetic resin in which a plurality of pores are provided in advance and arranging it on a substrate. Further, it may be used as a particle-like or fibrous cell culture scaffold material having a plurality of pores.

In the present invention, the zeta potential at pH 7 of the cell culture scaffold material is preferably −40 mV or more, more preferably −35 mV or more, still more preferably −30 mV or more, preferably + 30 mV or less, and more. It is preferably less than +30 mV, and more preferably less than +25 mV. In this case, the cell adhesion can be further enhanced.

The zeta potential of the cell culture scaffold material is a solution sufficiently separated from the surface of the cell culture scaffold material, which is the interface, and the surface of the cell culture scaffold material, when the cell culture scaffold material is placed in the solution. It is the potential difference from the bulk part inside. The value of the zeta potential in the present specification can be obtained by appropriately selecting a method such as a flow potential method, an electrophoresis method, or an electroosmosis method according to the shape of the scaffold material for cell culture.

For example, when the scaffold material for cell culture is a membrane, it can be measured by the flow potential method. When the scaffold material for cell culture is a fiber or a particle, it can be measured by an electroosmosis method.

The flow potential method can be measured by, for example, a zeta potential measuring device manufactured by Anton Pearl Co., Ltd. Specifically, it can be measured in a diluted solution of KCl by the method described in Examples described later. Examples of the electrophoresis method include a method of measuring by light scattering in water using a zeta potential / particle size / molecular weight measurement system (manufactured by Otsuka Electronics Co., Ltd.).

The zeta potential can be increased by increasing the content of a cationic functional group such as an amino group in the synthetic resin. Further, the zeta potential can be lowered by increasing the content of an anionic functional group such as a carboxyl group in the synthetic resin.

The cell culture scaffold material of the present invention has an elastic modulus at 25 ° C. measured by a nanoindentation method, preferably 0.01 GPa or more, more preferably 0.05 GPa or more, still more preferably 0.1 GPa or more, and particularly preferably 0.1 GPa or more. It is 0.5 GPa or more, preferably 5 GPa or less, more preferably 3 GPa or less, and further preferably 1 GPa or less. When the elastic modulus is within the above range, the cell adhesion can be further enhanced.

The elastic modulus can be measured using, for example, a nanoindenter (Hysitron, Tribonder, manufactured by Bruker). In this measurement, the indenter uses a Berkovic (triangular pyramid type) tip diameter R number of 100 nm, and a single indentation measurement is performed at room temperature in the atmosphere, and the indentation depth can be 50 nm.

Further, the scaffold material for cell culture of the present invention has a water swelling rate of preferably 100% or less, more preferably 50% or less, still more preferably 40% or less. In this case, the colonization of the cells after seeding can be further enhanced. The lower limit of the water swelling rate is not particularly limited, but may be, for example, 0.5%. In addition, the water swelling rate can be measured by, for example, the following method.

First, the scaffold material for cell culture is immersed in water at 25 ° C. for 24 hours. Next, the scaffold material for cell culture is taken out from the water and allowed to stand on the water absorbing material to discharge the water remaining in the pores. Next, the weights of the samples before and after immersion are measured, and the water swelling rate = (sample weight after immersion-sample weight before immersion) / (sample weight before immersion) × 100 (%) is calculated. ..

The water swelling rate can be reduced, for example, by increasing the hydrophobic functional groups of the synthetic resin, lowering the number average molecular weight, and the like.

[Synthetic resin]
The scaffold material for cell culture of the present invention contains a synthetic resin (hereinafter, may be referred to as synthetic resin X). In addition, in this specification, a "structural unit" means a repeating unit of a monomer constituting a synthetic resin. When the synthetic resin has a graft chain, it contains a repeating unit of the monomers constituting the graft chain.

The synthetic resin X preferably contains a Bronsted basic group in a constituent unit in an amount of 0.2 mol% or more, more preferably 2 mol% or more, and preferably 30 mol% or less. It is more preferably contained in an amount of mol% or less, and further preferably contained in an amount of 15 mol% or less. In this case, the effect of the present invention can be exhibited even more effectively. The Bronsted basic group and the like will be described later.

(Vinyl polymer)
The synthetic resin X is preferably a vinyl polymer. The vinyl polymer is a polymer of a compound having a vinyl group or a vinylidene group. When the synthetic resin X is a vinyl polymer, it is possible to more easily suppress the swelling of the scaffold material for cell culture in water. Examples of the vinyl polymer include polyvinyl alcohol derivatives, poly (meth) acrylic acid esters, polyvinylpyrrolidone, polystyrene, ethylene-vinyl acetate copolymers and the like. Further, the vinyl polymer is preferably a polyvinyl alcohol derivative or a poly (meth) acrylic acid ester from the viewpoint of easily enhancing the adhesiveness with cells.

(Synthetic resin having a polyvinyl acetal skeleton)
The cell culture scaffold material preferably contains a synthetic resin having a polyvinyl acetal skeleton. The synthetic resin X preferably contains a synthetic resin having a polyvinyl acetal skeleton.

In the present specification, "synthetic resin having a polyvinyl acetal skeleton" may be referred to as "polyvinyl acetal resin X".

The polyvinyl acetal resin X preferably has an acetal group, an acetyl group, and a hydroxyl group in the side chain. However, the polyvinyl acetal resin X does not have to have, for example, an acetyl group. For example, the polyvinyl acetal resin X does not have to have an acetyl group by binding all of the acetyl groups of the polyvinyl acetal resin X with a linker described later.

When synthesizing the polyvinyl acetal resin X, at least a step of acetalizing polyvinyl alcohol with an aldehyde is provided.

The aldehyde used for acetalizing polyvinyl alcohol for obtaining the polyvinyl acetal resin X is not particularly limited. Examples of the aldehyde include aldehydes having 1 to 10 carbon atoms. The aldehyde may have a chain aliphatic group, a cyclic aliphatic group or an aromatic group. The aldehyde may be a chain aldehyde or a cyclic aldehyde.

Examples of the above aldehydes include formaldehyde, acetaldehyde, propionaldehyde, butylaldehyde, pentanal, hexanal, heptanal, octanal, nonanal, decanal, achlorine, benzaldehyde, cinnamaldehyde, perylaldehyde, formylpyridine, formylimidazole, formylpyrrole, formylpiperidin, formyl. Triazole, formyltetrazole, formylindole, formylisoindole, formylpurine, formylbenzoimidazole, formylbenzotriazole, formylquinoline, formylisoquinolin, formylquinoxalin, formylcinnoline, formylpteridine, formylfuran, formyloxolane, formyloxane, Examples thereof include formylthiophene, formylthiolan, formyltian, formyladenine, formylguanine, formylcitosine, formyltimine, and formyluracil. Only one kind of the above aldehyde may be used, or two or more kinds may be used in combination.

The aldehyde is preferably formaldehyde, acetaldehyde, propionaldehyde, butyraldehyde, or pentanal, and more preferably butyraldehyde. Therefore, the polyvinyl acetal skeleton is preferably a polyvinyl butyral skeleton. The polyvinyl acetal resin X is preferably a synthetic resin having a polyvinyl butyral skeleton, and more preferably a polyvinyl butyral resin.

A vinyl compound may be copolymerized with the polyvinyl acetal resin X. That is, the polyvinyl acetal resin X may be a copolymer of a structural unit of the polyvinyl acetal resin and a vinyl compound. In the present invention, the polyvinyl acetal resin copolymerized with the vinyl compound is also referred to as a polyvinyl acetal resin.

The vinyl compound is a compound having a vinyl group (H ₂ C = CH−). The vinyl compound may be a polymer having a structural unit having a vinyl group.

The copolymer may be a block copolymer of a polyvinyl acetal resin and a vinyl compound, or may be a graft copolymer obtained by grafting a vinyl compound on the polyvinyl acetal resin. The copolymer is preferably a graft copolymer.

The copolymer can be synthesized, for example, by the following methods (1) to (3). (1) A method for synthesizing a polyvinyl acetal resin using polyvinyl alcohol in which a vinyl compound is copolymerized. (2) A method for synthesizing a polyvinyl acetal resin using polyvinyl alcohol and polyvinyl alcohol in which a vinyl compound is copolymerized. (3) A method of graft-copolymerizing a vinyl compound with a polyvinyl acetal resin before graft copolymerization.

Examples of the vinyl compound include ethylene, allylamine, vinylpyrrolidone, vinylimidazole, maleic anhydride, maleimide, itaconic acid, (meth) acrylic acid, vinylamine, and (meth) acrylic acid ester. Only one kind of these vinyl compounds may be used, or two or more kinds may be used in combination.

From the viewpoint of further enhancing the adhesiveness of cells, the polyvinyl acetal resin X preferably has a Bronsted basic group or a Bronsted acidic group, and more preferably has a Bronsted basic group. That is, it is preferable that a part of the polyvinyl acetal resin X is modified with a Bronsted basic group or a Bronsted acidic group, and a part of the polyvinyl acetal resin X is modified with a Bronsted basic group. preferable.

The polyvinyl acetal resin X preferably has a Bronsted basic group or a Bronsted acidic group in part, and more preferably has a Bronsted basic group. In that case, the cell adhesion can be further enhanced. The Bronsted basic group or Bronsted acidic group may be present in a part of the polyvinyl acetal resin X, in which case the monomer having the Bronsted basic group or Bronsted acidic group is copolymerized. It may be grafted. The degree of modification of the Bronsted basic group is preferably 0.2 mol% or more, more preferably 2 mol% or more, preferably 30 mol% or less, more preferably 20 mol% or less, still more preferably 15 mol%. It is as follows. If the degree of denaturation by the Bronsted basic group is within the above-mentioned specific range, the cell adhesion can be further enhanced.

Bronsted basic group is a general term for functional groups capable of receiving hydrogen ion H + from other substances. Examples of the blended basic group include amine-based basic groups such as a substituent having an imine structure, a substituent having an imide structure, a substituent having an amine structure, and a substituent having an amide structure. The blended basic group is not particularly limited, but is limited to a hydroxyamino group, a urea group, a conjugated amine-based functional group such as guanidine or biganide, piperazine, piperidine, pyrrolidine, 1,4-diazabicyclo [2.2.2] octane. , Hexamethylenetetraamine, morpholine, pyridine, pyridazine, pyrimidine, pyrazine, pyrrol, azatropylidene, pyridone, imidazole, benzoimidazole, benzotriazole, pyrazole, oxazole, imidazoline, triazole, thiazole, thiazine, tetrazole, indol, isoindole, purine. , Heterocyclic amino functional groups such as quinoline, isoquinolin, quinazoline, quinoxalin, cinnoline, pteridine, carbazole, aclysine, adenine, guanine, cytosine, timine, uracil, melamine, cyclic pyrrol functional groups such as porphyrin, chlorin, choline and Examples thereof include derivatives thereof.

Examples of the blended acidic group include a carboxyl group, a sulfonic acid group, a maleic acid group, a sulfinic acid group, a sulfenic acid group, a phosphoric acid group, a phosphonic acid group, and salts thereof. The Bronsted acidic group is preferably a carboxyl group.

The polyvinyl acetal resin X preferably has a structural unit having an imine structure, a structural unit having an imide structure, a structural unit having an amine structure, or a structural unit having an amide structure. In this case, it may have only one of these structural units, or it may have two or more.

The polyvinyl acetal resin X may have a structural unit having an imine structure. The imine structure refers to a structure having a C = N bond. In particular, the polyvinyl acetal resin X preferably has an imine structure in the side chain.

The polyvinyl acetal resin X may have a structural unit having an imide structure. The structural unit having an imide structure is preferably a structural unit having an imino group (= NH).

The polyvinyl acetal resin X preferably has an imino group in the side chain. In this case, the imino group may be directly bonded to the carbon atom constituting the main chain of the polyvinyl acetal resin X, or may be bonded to the main chain via a linking group such as an alkylene group.

The polyvinyl acetal resin X may have a structural unit having an amine structure. The amine group in the above amine structure may be a primary amine group, a secondary amine group, a tertiary amine group, or a quaternary amine group. Good.

The structural unit having an amine structure may be a structural unit having an amide structure. The amide structure refers to a structure having -C (= O) -NH-.

The polyvinyl acetal resin X preferably has an amine structure or an amide structure in the side chain. In this case, the amine structure or the amide structure may be directly bonded to the carbon atom constituting the main chain of the polyvinyl acetal resin X, or may be bonded to the main chain via a linking group such as an alkylene group.

The content of structural units having an imine structure, the content of structural units having an imide structure, the content of structural units having an amine structure, and the content of structural units having an amide structure are ¹ H-NMR (nuclear magnetism). It can be measured by the resonance spectrum).

(Synthetic resin having a poly (meth) acrylic acid ester skeleton)
The scaffold material for cell culture of the present invention preferably contains a synthetic resin having a poly (meth) acrylic acid ester skeleton. The synthetic resin X preferably contains a synthetic resin having a poly (meth) acrylic acid ester skeleton.

In the present specification, "synthetic resin having a poly (meth) acrylic acid ester skeleton" may be referred to as "poly (meth) acrylic acid ester resin X".

Therefore, the poly (meth) acrylic acid ester resin X is a resin having a poly (meth) acrylic acid ester skeleton.

The poly (meth) acrylic acid ester resin X is obtained by polymerizing the (meth) acrylic acid ester or by copolymerizing the (meth) acrylic acid ester with another monomer.

Examples of the (meth) acrylic acid ester include (meth) acrylic acid alkyl ester, (meth) acrylic acid cyclic alkyl ester, (meth) acrylic acid aryl ester, (meth) acrylamide, (meth) acrylic acid polyethylene glycol, and ( Meta) Phosphorylcholine acrylate and the like can be mentioned.

Examples of the (meth) acrylic acid alkyl ester include methyl (meth) acrylate, ethyl (meth) acrylate, n-propyl (meth) acrylate, isopropyl (meth) acrylate, n-butyl (meth) acrylate, and isobutyl (meth) acrylate. t-butyl (meth) acrylate, n-octyl (meth) acrylate, isooctyl (meth) acrylate, 2-ethylhexyl (meth) acrylate, nonyl (meth) acrylate, isononyl (meth) acrylate, decyl (meth) acrylate, isodecyl ( Examples thereof include meth) acrylate, lauryl (meth) acrylate, stearyl (meth) acrylate, and isotetradecyl (meth) acrylate.

The (meth) acrylic acid alkyl ester may be substituted with a substituent such as an alkoxy group having 1 to 3 carbon atoms and a tetrahydrofurfuryl group. Examples of such (meth) acrylic acid alkyl esters include methoxyethyl acrylate and tetrahydrofurfuryl acrylate.

Examples of the (meth) acrylic acid cyclic alkyl ester include cyclohexyl (meth) acrylate and isobornyl (meth) acrylate.

Examples of the (meth) acrylic acid aryl ester include phenyl (meth) acrylate and benzyl (meth) acrylate.

Examples of (meth) acrylamides include (meth) acrylamide, N-isopropyl (meth) acrylamide, N-tert-butyl (meth) acrylamide, N, N'-dimethyl (meth) acrylamide, and (3- (meth) acrylamide propyl). ) Trimethylammonium chloride, 4- (meth) acryloylmorpholine, 3- (meth) acryloyl-2-oxazolidinone, N- [3- (dimethylamino) propyl] (meth) acrylamide, N- (2-hydroxyethyl) (meth) ) Acrylamide, N-methylol (meth) acrylamide, 6- (meth) acrylamide hexane acid and the like.

Examples of polyethylene glycols (meth) acrylate include methoxy-polyethylene glycol (meth) acrylate, ethoxy-polyethylene glycol (meth) acrylate, hydroxy-polyethylene glycol (meth) acrylate, methoxy-diethylene glycol (meth) acrylate, and ethoxy-. Diethylene glycol (meth) acrylate, hydroxy-diethylene glycol (meth) acrylate, methoxy-triethylene glycol (meth) acrylate, ethoxy-triethylene glycol (meth) acrylate, hydroxy-triethylene glycol (meth) acrylate and the like can be mentioned. Be done.

Examples of phosphorylcholine (meth) acrylate include 2- (meth) acryloyloxyethyl phosphorylcholine.

As the other monomer copolymerized with the (meth) acrylic acid ester, a vinyl compound is preferably used. Examples of the vinyl compound include ethylene, allylamine, vinylpyrrolidone, maleic anhydride, maleimide, itaconic acid, (meth) acrylic acid, vinylamine, and (meth) acrylic acid ester. Only one kind of vinyl compound may be used, or two or more kinds may be used in combination.

In addition, in this specification, "(meth) acrylic" means "acrylic" or "methacryl", and "(meth) acrylate" means "acrylate" or "methacrylate".

Like the polyvinyl acetal resin X, the poly (meth) acrylic acid ester resin X preferably has a Bronsted basic group or a Bronsted acidic group in part. In that case, the extensibility can be further enhanced. The Bronsted basic group may be present in a part of the poly (meth) acrylic acid ester resin X, and in that case, the monomer having the Bronsted basic group may be copolymerized, and the graft may be formed. It may have been. The degree of modification of the Bronsted basic group is preferably 0.2 mol% or more, more preferably 2 mol% or more, preferably 30 mol% or less, more preferably 20 mol% or less, still more preferably 15 mol%. It is as follows. If the degree of denaturation by the Bronsted basic group is within the above-mentioned specific range, the adhesion of cells can be further enhanced.

(Synthetic resin having a peptide part)
The scaffold material for cell culture of the present invention preferably contains a synthetic resin having a peptide portion. The synthetic resin X preferably contains a synthetic resin having a peptide portion. By using a synthetic resin having a peptide portion as a material for a scaffold material for cell culture, the fixability after seeding of cells is further improved, and the proliferation rate of cells can be further increased.

A synthetic resin having a peptide portion can be obtained by reacting a synthetic resin with a linker and a peptide. The synthetic resin having a peptide portion is preferably a peptide-containing polyvinyl acetal resin having a polyvinyl acetal resin portion, a linker portion, and a peptide portion, and a peptide having a polyvinyl butyral resin portion, a linker portion, and a peptide portion. More preferably, it is a containing polyvinyl butyral resin. Therefore, the synthetic resin having the polyvinyl acetal skeleton (polyvinyl acetal resin X) is preferably a peptide-containing polyvinyl acetal resin, and more preferably a peptide-containing polyvinyl butyral resin. As the synthetic resin having a peptide portion, only one kind may be used, or two or more kinds may be used in combination.

The peptide portion is preferably composed of 3 or more amino acids, more preferably composed of 4 or more amino acids, further preferably composed of 5 or more amino acids, and 10 or less. It is preferably composed of amino acids, and more preferably composed of 6 or less amino acids. When the number of amino acids constituting the peptide portion is not less than the above lower limit and not more than the above upper limit, the adhesiveness to the cells after seeding can be further enhanced, and the cell proliferation rate can be further enhanced.

The peptide portion preferably has a cell-adhesive amino acid sequence. The cell adhesion amino acid sequence refers to an amino acid sequence whose cell adhesion activity has been confirmed by the phage display method, the Sepharose beads method, or the plate coating method. As the phage display method, for example, the method described in "The Journal of Cell Biology, Volume 130, Number 5, September 1995 1189-1196" can be used. As the Sepharose beads method, for example, the method described in "Protein Nucleic Acid Enzyme Vol. 45 No. 15 (2000) 2477" can be used. As the plate coating method, for example, the method described in "Protein Nucleic Acid Enzyme Vol. 45 No. 15 (2000) 2477" can be used.

Examples of the cell-adhesive amino acid sequence include an RGD sequence (Arg-Gly-Asp), a YIGSR sequence (Tyr-Ile-Gly-Ser-Arg), and a PDSGR sequence (Pro-Asp-Ser-Gly-Arg). HAV sequence (His-Ala-Val), ADT sequence (Ala-Asp-Thr), QAV sequence (Gln-Ala-Val), LDV sequence (Leu-Asp-Val), IDS sequence (Ile-Asp-Ser), REDV sequence (Arg-Glu-Asp-Val), IDAPS sequence (Ile-Asp-Ala-Pro-Ser), KQAGDV sequence (Lys-Gln-Ala-Gly-Asp-Val), and TDE sequence (Thr-Asp- Glu) and the like. The cell-adherent amino acid sequences include "Physiology of Pathology, Vol. 9, No. 7, pp. 527-535, 1990" and "Osaka Prefectural Medical Center for Maternal and Child Health, Vol. 8, No. 1, 58-". The sequences described in "page 66, 1992" are also mentioned. The peptide portion may have only one type of the cell-adhesive amino acid sequence, or may have two or more types.

The cell-adherent amino acid sequence preferably has at least one of the cell-adherent amino acid sequences described above, and more preferably has at least an RGD sequence, a YIGSR sequence, or a PDSGR sequence, and the following formula (1) It is more preferable to have at least the RGD sequence represented by). In this case, the adhesiveness to the cells after seeding can be further enhanced, and the cell proliferation rate can be further enhanced.

Arg-Gly-Asp-X ... Equation (1)

In the above formula (1), X represents Gly, Ala, Val, Ser, Thr, Phe, Met, Pro, or Asn.

The peptide portion may be linear or may have a cyclic peptide skeleton. The cyclic peptide skeleton is a cyclic skeleton composed of a plurality of amino acids. From the viewpoint of exerting the effect of the present invention more effectively, the cyclic peptide skeleton is preferably composed of 4 or more amino acids, more preferably 5 or more amino acids, and 10 or more. It is preferably composed of the following amino acids.

In the synthetic resin having a peptide portion, the content of the peptide portion is preferably 0.1 mol% or more, more preferably 1 mol% or more, still more preferably 5 mol% or more, and particularly preferably 10 mol% or more. .. In the synthetic resin having a peptide portion, the content of the peptide portion is preferably 60 mol% or less, more preferably 50 mol% or less, still more preferably 35 mol% or less, and particularly preferably 25 mol% or less. When the content of the peptide portion is within the above range, the adhesiveness to the cells after seeding can be further enhanced, and the proliferation rate of the cells can be further enhanced. Further, when the content of the peptide portion is not more than the above upper limit, the production cost can be suppressed. The content rate (mol%) of the peptide portion is the amount of substance of the peptide portion with respect to the total amount of substance of each structural unit constituting the synthetic resin having the peptide portion.

The content of the peptide portion can be measured by FT-IR or LC-MS.

In a synthetic resin having a peptide portion, it is preferable that the synthetic resin portion and the peptide portion are bonded via a linker. That is, the synthetic resin having a peptide portion is preferably a synthetic resin having a peptide portion and a linker portion. Only one type of the linker may be used, or two or more types may be used in combination.

The linker is preferably a compound having a functional group capable of condensing with the carboxyl group or amino group of the peptide. Examples of the functional group capable of condensing with the carboxyl group or amino group of the peptide include a carboxyl group, a thiol group and an amino group. From the viewpoint of good reaction with the peptide, the linker is preferably a compound having a carboxyl group. As the linker, the vinyl compound described above can also be used.

Examples of the linker having a carboxyl group include (meth) acrylic acid and acrylamide containing a carboxyl group. By using a carboxylic acid (carboxylic acid monomer) having a polymerizable unsaturated group as the linker having a carboxyl group, the carboxylic acid monomer can be polymerized by graft polymerization when the linker and the synthetic resin are reacted. Therefore, the number of carboxyl groups that can be reacted with the peptide can be increased.

(Other resins)
The scaffold material for cell culture may contain a polymer other than the synthetic resin described above. Examples of the polymer include polyvinylpyrrolidone, polystyrene, ethylene / vinyl acetate copolymer, polyolefin resin, polyether resin, polyvinyl alcohol resin, polyester, epoxy resin, polyamide resin, polyimide resin, polyurethane resin, polycarbonate resin and the like.

[Other details of scaffolding material for cell culture]
The cell culture scaffold material according to the present invention contains the above synthetic resin X. From the viewpoint of effectively exerting the effect of the present invention and increasing productivity, the content of the synthetic resin X in 100% by weight of the scaffolding material for cell culture is preferably 90% by weight or more, more preferably 90% by weight or more. It is 95% by weight or more, more preferably 97.5% by weight or more, particularly preferably 99% by weight or more, and most preferably 100% by weight (total amount). Therefore, it is most preferable that the scaffold material for cell culture is the synthetic resin X. When the content of the synthetic resin X is at least the above lower limit, the effect of the present invention can be exhibited even more effectively.

The cell culture scaffold material may contain components other than the synthetic resin X. Examples of the components other than the synthetic resin X include polysaccharides, celluloses, synthetic peptides and the like.

From the viewpoint of effectively exerting the effects of the present invention, the smaller the content of the components other than the synthetic resin X, the better. The content of the component in 100% by weight of the scaffold material for cell culture is preferably 10% by weight or less, more preferably 5% by weight or less, still more preferably 2.5% by weight or less, and particularly preferably 1% by weight or less. , Most preferably 0% by weight (not contained). Therefore, it is most preferable that the scaffold material for cell culture does not contain components other than the synthetic resin X.

The cell culture scaffold material according to the present invention preferably does not contain animal-derived raw materials. By not containing animal-derived raw materials, it is possible to provide a scaffold material for cell culture that is highly safe and has little variation in quality during production.

(Cell culture using scaffolding material for cell culture)
The cell culture scaffold material according to the present invention is used for culturing cells. The cell culture scaffold material according to the present invention is used as a scaffold for the cells when culturing the cells.

Examples of the cells include animal cells such as humans, mice, rats, pigs, cows and monkeys. Examples of the cells include somatic cells and the like, and examples thereof include stem cells, progenitor cells and mature cells. The somatic cells may be cancer cells.

Examples of the stem cells include mesenchymal stem cells (MSCs), iPS cells, ES cells, Muse cells, embryonic cancer cells, embryonic reproductive stem cells, mGS cells and the like.

Examples of the mature cells include nerve cells, cardiomyocytes, retinal cells, hepatocytes and the like.

(Shape of scaffolding material for cell culture)
The shape of the scaffold material for cell culture of the present invention is not particularly limited, and may be particles, fibers, porous bodies, or films. The particles, fibers, porous body, or film may contain components other than the scaffold material for cell culture.

When the shape of the scaffold material for cell culture is a film, it is preferably used for plane culture (two-dimensional culture) of cells. When the shape of the scaffold material for cell culture is particles, fibers, or a porous body, it is preferably used for three-dimensional culture of cells.

(Cell culture container)
The present invention also relates to a cell culture vessel in which the cell culture scaffold material is provided in at least a part of the cell culture region. FIG. 1 is a schematic front sectional view showing a cell culture container according to an embodiment of the present invention.

The cell culture container 1 includes a container body 2 and a cell culture scaffold material 3. The cell culture scaffold material 3 is arranged on the surface 2a of the container body 2. A scaffold material 3 for cell culture is arranged on the bottom surface of the container body 2. By adding a liquid medium to the cell culture container 1 and seeding the cells on the surface of the cell culture scaffold material 3, the cells can be cultured in a plane or three-dimensionally.

The container body may be provided with a second container body such as a cover glass on the bottom surface of the first container body. The first container body and the second container body may be separable. In this case, the cell culture scaffold material may be arranged on the surface of the second container body.

As the container body, a conventionally known container body (container) can be used. The shape and size of the container body are not particularly limited.

Examples of the container body include a cell culture plate provided with one or more wells (holes), a cell culture flask, and the like. The number of wells in the plate is not particularly limited. The number of wells is not particularly limited, and examples thereof include 2, 4, 6, 12, 24, 48, 96, and 384. The shape of the well is not particularly limited, and examples thereof include a perfect circle, an ellipse, a triangle, a square, a rectangle, and a pentagon. The shape of the bottom surface of the well is not particularly limited, and examples thereof include a flat bottom, a round bottom, and unevenness.

The material of the container body is not particularly limited, and examples thereof include resin, metal, and inorganic materials. Examples of the resin include polystyrene, polyethylene, polypropylene, polycarbonate, polyester, polyisoprene, cycloolefin polymer, polyimide, polyamide, polyamideimide, (meth) acrylic resin, epoxy resin, silicone and the like. Examples of the metal include stainless steel, copper, iron, nickel, aluminum, titanium, gold, silver, platinum and the like. Examples of the inorganic material include silicon oxide (glass), aluminum oxide, titanium oxide, zirconium oxide, iron oxide, and silicon nitride.

Next, the present invention will be clarified by giving specific examples and comparative examples of the present invention. The present invention is not limited to the following examples.

The following synthetic resin X1 was synthesized as a raw material for a scaffold material for cell culture.

300 parts by weight of amine-modified polyvinyl alcohol having an ion-exchanged water of 2700 mL, an average degree of polymerization of 800, an amine modification degree of 1 mol%, and a saponification degree of 97.5 mol% was put into a reactor equipped with a stirrer and dissolved by heating while stirring. And obtained a solution. To the obtained solution, 35% by weight hydrochloric acid was added as a catalyst so that the hydrochloric acid concentration was 0.2% by weight. Then, the temperature was adjusted to 15 ° C., and 22 parts by weight of n-butyraldehyde was added with stirring. Then, 148 parts by weight of n-butyraldehyde was added to precipitate a white particulate polyvinyl butyral resin. Fifteen minutes after the precipitation, 35% by weight hydrochloric acid was added so that the hydrochloric acid concentration became 1.8% by weight, and then the mixture was heated to 50 ° C. and maintained at 50 ° C. for 2 hours. Then, the solution was cooled and neutralized, and then the polyvinyl butyral resin was washed with water and dried to obtain a synthetic resin X1 which is a polyvinyl butyral resin.

The obtained polyvinyl butyral resin (synthetic resin X1) had an average degree of polymerization of 800, a hydroxyl group amount of 20 mol%, an amine group amount of 1 mol%, an acetylation degree of 1 mol%, and a butyralization degree of 78 mol%.

The content of structural units in the obtained synthetic resin was ^{measured by 1} H-NMR (nuclear magnetic resonance spectrum) after dissolving the synthetic resin in DMSO-D6 (dimethylsulfoxide).

(Examples 1 to 10 and Comparative Examples 2 and 3)
In Examples 1 and 2, the synthetic resin X1 was used, and in Examples 3 to 7, 9 and 10, synthetic resins X2 to X8 were used, respectively.

The synthetic resin X2 was obtained by blending 5 mol% of dimethylaminoacrylamide as a monomer with polyvinyl butyral resin having a butyralization degree of 68 mol%, an acetylation degree of 1 mol% and a hydroxyl group amount of 26 mol%. The synthesis conditions were as follows. That is, the above polyvinyl butyral resin was dissolved in THF at a concentration of 20% by weight, 5 parts by weight of dimethylaminoacrylamide was added, and then 3 parts by weight of Irgacure184 (manufactured by IGM Resins BV) was dissolved as an initiator to UV. UV irradiation was performed for 20 minutes in a polymerization tester.

Further, the synthetic resin X3 was obtained in the same manner as the synthetic resin X2 except that the content ratio of dimethylaminoacrylamide was changed from 5 mol% to 20 mol% as shown in Table 1.

The synthetic resin X4 was obtained in the same manner as the synthetic resin X2 except that diethylaminoacrylamide was used as a monomer and the content ratio was 28 mol%.

The synthetic resin X5 was obtained in the same manner as the synthetic resin X2 except that vinyl imidazole was used as a monomer and the content ratio was 13 mol%.

The synthetic resin X6 is the same as in Example 1 except that polyvinyl alcohol having an average degree of polymerization of 2400 and a saponification degree of 97 mol% is used, and acetaldehyde is used instead of n-butyraldehyde (n-BA). A synthetic resin, which is a polyvinyl acetal resin, was obtained. It was obtained by blending 5 mol% of dimethylaminoacrylamide as a monomer with the obtained polyvinyl acetal resin. The synthesis conditions were as follows. That is, the obtained polyvinyl acetal resin was dissolved in THF at a concentration of 20% by weight, 5 parts by weight of dimethylaminoacrylamide was added, and then 3 parts by weight of the initiator (Irgacure184) was dissolved, and the UV polymerization tester was used for 20 minutes. UV irradiation was performed.

The synthetic resin X7 was produced as follows. First, 81.4 parts by weight of butyl methacrylate and 18.6 parts by weight of hydroxyethyl methacrylate were mixed to obtain a (meth) acrylic monomer solution. Next, 0.5 part by weight of Irgacure 184 (manufactured by BASF) was dissolved in the obtained (meth) acrylic monomer solution and applied onto a PET film. A poly (meth) acrylic acid ester resin solution is obtained by irradiating the coated material at 25 ° C. with a UV conveyor device "ECS301G1" manufactured by Eye Graphics Co., Ltd. with a wavelength of 365 nm at an integrated light intensity of 2000 mJ / cm ^2. It was. The obtained poly (meth) acrylic acid ester resin solution was vacuum-dried at 80 ° C. for 3 hours to obtain a synthetic resin X7 as a poly (meth) acrylic acid ester resin.

In the synthetic resin X8, a synthetic resin which is a polyvinyl acetal resin was obtained in the same manner as in Example 1 except that polyvinyl alcohol having an average degree of polymerization of 800 and a saponification degree of 1 mol% was used. 95 parts by weight of the obtained polyvinyl acetal resin and 5 parts by weight of acrylic acid (linker) are dissolved in 300 parts by weight of THF and reacted in the presence of a photoradical polymerization initiator for 20 minutes under ultraviolet irradiation to obtain a polyvinyl acetal resin. And acrylic acid were graft-copolymerized to form a linker portion.

As a peptide, a linear peptide having an amino acid sequence of Gly-Arg-Gly-Asp-Ser (five amino acid residues, described as GRGDS in Table 1) was prepared. 5 parts by weight of this peptide and 5 parts by weight of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (condensing agent) were added to 90 parts by weight of methanol containing neither calcium nor magnesium, and the peptide was added. A containing solution was prepared. 100 parts by weight of this peptide-containing liquid and 97 parts by weight of the polyvinyl acetal resin having the linker portion formed are added to 300 parts by weight of methanol and reacted to dehydrate and condense the carboxyl group of the linker portion and the amino group of Arg of the peptide. did. After completion of the reaction, vacuum drying was performed at 60 ° C. for 1 hour to volatilize methanol. In this way, a synthetic resin X8 which is a peptide-containing polyvinyl acetal resin having a polyvinyl acetal resin portion, a linker portion, and a peptide portion was produced.

Further, in Comparative Example 2, synthetic resin X1 was used, and in Example 8 and Comparative Example 3, a polyvinyl butyral resin having the composition shown in Table 1 was used.

Preparation of cell culture container;
(Examples 1 to 10, Comparative Example 3)
In Examples 1 to 10 and Comparative Example 3, first, the synthetic resin shown in Table 1 was dissolved in a butanol / methanol mixed solvent (9: 1) at a concentration of 5 wt%. 40 μL of the obtained solution was cast into a polystyrene dish having a diameter of 22 mm, immersed in liquid nitrogen, and frozen together with the solvent. Then, it was freeze-dried for 2 days using a freeze-dryer to obtain a cell culture container provided with a cell culture scaffold material having pores.

The average diameter of the pores of the cell culture scaffold material was determined using a scanning electron microscope (SEM). FIG. 2 is an SEM observation image of the cell culture scaffold material obtained in Example 1 at a magnification of 1000 times, and FIG. 3 is an SEM observation image of the cell culture scaffold material obtained in Example 1 at a magnification of 10000 times. It is an SEM observation image.

(Comparative Examples 1 and 2)
In Comparative Example 1, the polystyrene dish itself was used as a cell culture container.

In Comparative Example 2, first, the synthetic resin X1 was dissolved in a butanol / methanol mixed solvent (9: 1) at a concentration of 5 wt%. 40 μL of the obtained solution was cast on a polystyrene dish having a diameter of 22 mm and then dried in an oven at 45 ° C. for 6 hours without freezing to form a flat film. In Comparative Examples 1 and 2, there were no pores whose average diameter could be measured.

Measurement of zeta potential;
Using a zeta potential measuring device (model number SURPASS 3, manufactured by Anton Pearl Co., Ltd.), the zeta potentials at pH 7 of the cell culture scaffold materials of Examples 1 to 10 and Comparative Examples 1 to 3 were determined, respectively. First, a 0.1 mM KCl solution was prepared. Next, after setting the resin film of the scaffolding material in the cell, the pH electrode and the conductivity meter were set. The zeta potential at each pH was measured by titrating to pH 9-3 using HCl and NaOH solution.

In Comparative Example 3, the zeta potential was not more than the lower limit of measurement, so that the measurement was impossible.

Water swelling rate;
When determining the water swelling rate of each scaffold material, the scaffold material was taken out from the cell culture container and immersed in water at 25 ° C. for 24 hours. The sample after immersion was once allowed to stand on a Kim towel for 1 minute to drain water in the pores, and then weighed. The weights of the samples before and after immersion were measured, and the water swelling rate = (sample weight after immersion-sample weight before immersion) / (sample weight before immersion) × 100 (%) was calculated.

Surface elastic modulus measurement;
The surface elastic modulus of the cell culture scaffold materials of Examples 1 to 10 and Comparative Examples 1 to 3 at 25 ° C. was determined using a nanoindenter (Hysitron, Triboinder, manufactured by Bruker). As the indenter, a Berkovic (triangular pyramid type) tip diameter R number of 100 nm was used, and a single indentation measurement was performed at room temperature in the atmosphere, and the indentation depth at that time was 50 nm.

Average diameter of pores:
The surface of the scaffold material for cell culture was observed using a scanning electron microscope (SEM), and the average value of the pore sizes of 100 arbitrarily selected pores in the observation region (5 μm × 5 μm) was determined.

Seeding and culturing cells;
The following liquid medium and ROCK (Rho-associated kinase) -specific inhibitor were prepared.

TeSR E8 medium (manufactured by STEM CELL)
ROCK-Inhibitor (Y27632)

1 mL of phosphate buffered saline was added to the obtained cell culture vessel, and the mixture was allowed to stand in an incubator at 37 ° C. for 1 hour, and then the phosphate buffered saline was removed from the cell culture vessel.

Colonies of h-iPS cells 253G1 in a confluent state were placed on a dish having a diameter of 35 mm, 1 mL of 0.5 mM ethylenediamine / phosphate buffer solution was added, and the mixture was allowed to stand at room temperature for 2 minutes. After removing the ethylenediamine / phosphate buffer solution, the cells were pipetted with 1 mL of TeSR E8 medium to obtain cell clusters crushed to a size of 50 μm to 200 μm. The obtained cell mass (cell number 0.2 × 105 cells) was clamp-seeded in the above cell culture container.

At the time of seeding, 1.5 mL of liquid medium and a ROCK-specific inhibitor were added to a cell culture vessel so that the final concentration was 10 μM, and the cells were cultured in _{an incubator at 37 ° C. and a CO 2} concentration of 5%. .. Then, the work of exchanging the liquid in the dish with 1.5 mL of fresh liquid medium was repeated every 24 hours, and the cells were cultured for 5 days. After adding 1 mL of Triple Express manufactured by Thermo-Fisher to the dish after culturing and allowing it to stand at 37 ° C. for 2 minutes, the cells were collected and the number of cells was measured using the auto cell counter EVE.

The results are shown in Table 1 below.

1 ... Cell culture container 2 ... Container body 2a ... Surface 3 ... Cell culture scaffolding material

Claims (11)

Contains synthetic resin
The zeta potential at pH 7 is -60 mV or higher,
A scaffold material for cell culture having a plurality of pores having an average diameter of 10 nm or more and 5 μm or less. The scaffold material for cell culture according to claim 1, wherein the zeta potential at pH 7 is +30 mV or less. The scaffold material for cell culture according to claim 1 or 2, wherein the elastic modulus at 25 ° C. measured by the nanoindentation method is 0.01 GPa or more. The scaffold material for cell culture according to any one of claims 1 to 3, wherein the synthetic resin contains 0.2 mol% or more and 20 mol% or less of Bronsted basic groups in the structural unit. The scaffold material for cell culture according to any one of claims 1 to 4, wherein the water swelling rate is 100% or less. The scaffold material for cell culture according to any one of claims 1 to 5, which is substantially free of animal-derived raw materials. The scaffold material for cell culture according to any one of claims 1 to 6, wherein the synthetic resin contains a vinyl polymer. The scaffold material for cell culture according to any one of claims 1 to 7, wherein the synthetic resin contains at least a polyvinyl alcohol derivative or a poly (meth) acrylic resin. The scaffold material for cell culture according to any one of claims 1 to 8, wherein the synthetic resin contains a polyvinyl acetal resin. The scaffold material for cell culture according to any one of claims 1 to 9, wherein the number of pores per unit area is 0.01 cells / μm ² or more and 1000 cells / μm ^{2 or less.} A cell culture container comprising the cell culture scaffold material according to any one of claims 1 to 10 in at least a part of the cell culture region.