JP7070543B2 - Radiation-sensitive resin composition, pattern membrane and its manufacturing method, pattern substrate, cell culture instrument, microchannel device, and cell mass manufacturing method. - Google Patents

Description

The present invention relates to a radiation-sensitive resin composition, a pattern membrane and a method for producing the same, a pattern substrate, a cell culture device, a microchannel device, and a method for producing a cell mass.

In cell culture technology, attention is being paid to the culture of tissues having the same functions as in vivo tissues. The tissue is, for example, a cell mass formed by sterically gathering a large number of cells (see, for example, Patent Documents 1 and 2).

International Publication No. 2007/097120 Japanese Unexamined Patent Publication No. 2006-12191

As a cell culture technique, a patterned membrane can be formed using a radiation-sensitive resin composition, and the patterned membrane can be used as a partition wall to culture cells in a region defined by the partition wall. Here, for example, when a radiation-sensitive acid generator or base generator is used for pattern formation in the resin composition, the acid or base generated during pattern formation adversely affects the cells, for example, the aggregation of the cells deteriorates. There is a possibility of adverse effects such as

The present invention provides a radiation-sensitive resin composition capable of suppressing the adverse effects of acids or bases generated during pattern formation on cells, a method for producing a pattern film using the composition, and a pattern film. That is the issue. Another object of the present invention is to provide a pattern substrate, a cell culture device, and a microchannel device having excellent cell adhesion prevention properties. Another object of the present invention is to provide a method for producing a cell mass.

The present inventors have made diligent studies to solve the above problems. As a result, they have found that the above problems can be solved by the following configurations, and have completed the present invention. The present invention relates to, for example, the following [1] to [14].

[1] A polymer (A) having a structural unit (I) represented by the formula (1), and at least one radiation-sensitive compound selected from a radiation-sensitive acid generator and a radiation-sensitive base generator ( B) and a radiation-sensitive resin composition containing.

［式（１）中、Ｒ¹は、水素原子、メチル基またはトリフルオロメチル基であり、Ｒ²およびＲ³は、それぞれ独立に炭素数１～５のアルキル基またはアリール基であり、Ｒ⁴およびＲ⁵は、それぞれ独立に炭素数１～１０のアルカンジイル基であり、Ｘは、カルボアニオン（－ＣＯＯ^-基）、スルホアニオン（－ＳＯ₃ ^-基）、またはリン酸アニオンであり、Ｑは、酸素原子、エステル結合、アミド結合、アリーレン基、炭素数１～１０のアルカンジイル基、またはこれらの基の組合せである。］

[In formula (1), R ¹ is a hydrogen atom, a methyl group or a trifluoromethyl group, and R ² and R ³ are independently alkyl or aryl groups having 1 to 5 carbon atoms, respectively, and R ⁴ And R ⁵ are independently alcandiyl groups having 1 to 10 carbon atoms, and X is a carboanion (-COO ^- group), a sulfoanion (-SO ₃ ^- group), or a phosphate anion, and Q. Is an oxygen atom, an ester bond, an amide bond, an arylene group, an alcandiyl group having 1 to 10 carbon atoms, or a combination thereof. ]

[2] The radiation-sensitive resin composition according to the above [1], wherein the polymer (A) further has a structural unit (II) containing at least one selected from a cyclic ether structure and a cyclic carbonate structure.
[3] The radiation-sensitive resin composition according to the above [2], wherein the structural unit (II) contains a cyclic ether structure, and the cyclic ether structure is at least one selected from an oxylan structure and an oxetane structure.

[4] The item according to any one of [1] to [3] above, wherein the content ratio of the structural unit (I) is 10 to 70 mol% in 100 mol% of the total structural unit of the polymer (A). Radiation sensitive resin composition.
[5] The radiation-sensitive radiation according to any one of the above [1] to [4], which contains the radiation-sensitive compound (B) in a range of 10 parts by mass or less with respect to 100 parts by mass of the polymer (A). Sex resin composition.

[6] The radiation-sensitive resin composition according to any one of [1] to [5] above, which further contains a solvent having an octanol / water partition coefficient of 0 or less.
[7] (1) A step of forming a resin film of the radiation-sensitive resin composition according to any one of the above [1] to [6] on a substrate, (2) a step of exposing the resin film. (3) A method for producing a pattern film, which comprises a step of developing the exposed resin film with a developing solution.

[8] The method for producing a pattern film according to the above [7], wherein the developer used in the step (3) contains water.
[9] The method for producing a pattern film according to the above [7] or [8], wherein the steps (1) to (3) are performed at a temperature of 200 ° C. or lower.

[10] A pattern film formed by the method for producing a pattern film according to any one of the above [7] to [9].
[11] A cell culture device having the pattern membrane according to the above [10] on a part of the surface.

[12] A microchannel device having the pattern film according to the above [10] on a part of the inner surface of the microchannel.
[13] A pattern substrate having a substrate and a pattern film on the substrate, and the size of the non-existent portion of the film and the thickness of the pattern film are independently 0.1 μm to 1 mm, respectively. A pattern substrate in which the pattern film contains a polymer having the structural unit (I) represented by the formula (1).
[14] A method for producing a cell mass, which comprises a step of culturing cells using the cell culture device according to the above [11] to form a cell mass.

According to the present invention, a radiation-sensitive resin composition capable of suppressing the adverse effect of an acid or a base generated during pattern formation on cells, a method for producing a pattern film using the composition, and a pattern film are provided. Can be provided. The composition has excellent photolithography performance, can form a pattern film having an excellent effect of preventing cell adhesion, and can selectively aggregate cells in a desired portion.

Further, according to the present invention, it is possible to provide a pattern substrate, a cell culture device and a microchannel device having excellent cell adhesion prevention properties, and it is also possible to provide a method for producing a cell mass.

Hereinafter, embodiments for carrying out the present invention will be described.
[Radiation-sensitive resin composition]
The radiation-sensitive resin composition of the present invention (also simply referred to as “resin composition”) is at least one selected from the polymer (A) described below, a radiation-sensitive acid generator and a radiation-sensitive base generator. Contains a species of radiation-sensitive compound (B).

<Polymer (A)>
<Structural unit (I)>
The polymer (A) has a structural unit (I) represented by the formula (1). The polymer (A) may have one type of structural unit (I) or two or more types of structural unit (I).

細胞培養用途のパターン膜等の材料として感放射線性組成物を用いる場合、パターン形成時に酸発生剤または塩基発生剤から発生する酸または塩基は、細胞集合性が悪化する等、細胞に悪影響を及ぼす可能性がある。しかしながら、本発明では構造単位（Ｉ）というベタイン構造を有する重合体（Ａ）が、前記酸または塩基に対してクエンチャーの役割を果たし、これらをトラップすると考えられ、したがって細胞への前記悪影響が抑制されているものと推測される。また、構造単位（Ｉ）を有する重合体（Ａ）は水溶性が高く、水現像性に優れている。

When a radiation-sensitive composition is used as a material for a pattern membrane or the like for cell culture, the acid or base generated from the acid generator or the base generator at the time of pattern formation has an adverse effect on cells such as deterioration of cell aggregation. there is a possibility. However, in the present invention, the polymer (A) having a betaine structure called the structural unit (I) is considered to act as a quencher for the acid or base and trap them, and thus the adverse effect on the cells is considered. It is presumed that it is suppressed. Further, the polymer (A) having the structural unit (I) has high water solubility and is excellent in water developability.

In the present specification, the betaine structure is an atom having a positive charge and a negative charge at positions not adjacent to each other (R ⁵ intervenes in the equation (1)) and has a positive formal charge (formula (1)). Then, it means a structure in which a dissociable hydrogen atom is not bonded to N atom).

In the formula (1), R ¹ is a hydrogen atom, a methyl group or a trifluoromethyl group, and is preferably a methyl group because a polymerizable compound for deriving the structural unit (I) is easily available. R ² and R ³ are independently alkyl or aryl groups having 1 to 5 carbon atoms, and are preferably alkyl groups. R ⁴ and R ⁵ are each independently an alkanediyl group having 1 to 10 carbon atoms.

X is a _carboanion (-COO ^- group), a sulfoanion (-SO3 ^- group), or a phosphate anion, preferably a carboanion or a sulfoanion.
Q is an oxygen atom (-O-), an ester bond (-COO-), an amide bond (-CONR-; R is a hydrogen atom or an alkyl group having 1 to 10 carbon atoms), an arylene group, and 1 to 10 carbon atoms. It is an alcandiyl group or a combination of these groups (that is, an oxygen atom, an ester bond, an amide bond, an arylene group, an alcandiyl group having 1 to 10 carbon atoms), and is preferably an ester bond or an amide bond.

The alkyl group in R ² and R ³ is preferably an alkyl group having 1 to 3 carbon atoms, and examples thereof include a methyl group, an ethyl group and a propyl group, and a methyl group is preferable. The aryl group in R ² and R ³ is preferably an aryl group having 6 to 10 carbon atoms, and examples thereof include a phenyl group, a tolyl group, and a naphthyl group, and a phenyl group is preferable.

The alkanediyl group in R ⁴ and R ⁵ is preferably an alkanediyl group having 1 to 5 carbon atoms. The alkanediyl group is preferably a linear alkanediyl group, and examples thereof include a methanediyl group, an ethane-1,2-diyl group, and a propane-1,3-diyl group.

The arylene group in Q is preferably an arylene group having 6 to 10 carbon atoms, and examples thereof include a phenylene group, a trilene group, and a naphthylene group. The alkanediyl group in Q is preferably an alkanediyl group having 1 to 5 carbon atoms, and examples thereof include a methanediyl group, an ethane-1,2-diyl group, and a propane-1,3-diyl group.

Examples of the polymerizable compound that derives the structural unit (I) include N- (meth) acryloyloxyethyl-N, N-dimethylammonium-α-N-methylcarboxybetaine, and N- (meth) acryloyloxyethyl-N. N-dimethylammonium-α-N-ethylcarboxybetaine, N- (meth) acryloyloxyethyl-N, N-dimethylammonium-α-N-propylsulfobetaine, N- (meth) acryloylaminopropyl-N, N- Included is dimethylammonium-α-N-propylsulfobetaine.
The following examples are particularly preferable for the structural unit (I).

<Structural unit (II)>
The polymer (A) contains a structural unit (II) containing at least one selected from a cyclic ether structure and a cyclic carbonate structure from the viewpoint of using the radiation-sensitive resin composition as a negative material and the cross-linking reactivity. It is preferable to have more. The cyclic ether structure is preferably at least one selected from the oxylan structure and the oxetane structure.
The polymer (A) can have one or more structural units (II).
The structural unit (II) is preferably represented by the formula (2).

式（２）中、Ｒ⁶は、水素原子、メチル基またはトリフルオロメチル基であり、好ましくはメチル基である。Ｒ⁷は、炭素数１～１０のアルカンジイル基である。Ｑは、酸素原子、エステル結合、アミド結合、アリーレン基、炭素数１～１０のアルカンジイル基、またはこれらの基（すなわち酸素原子、エステル結合、アミド結合、アリーレン基、炭素数１～１０のアルカンジイル基）の組合せである。Ｒ⁷およびＱにおける各基の具体例は、式（１）中のＲ⁴、Ｒ⁵およびＱで説明した各基の具体例と同様である。前記基の組合せとしては、例えば、アリーレンオキシ基（－Ａｒ－Ｏ－；Ａｒはアリーレン基）、アラルキレンオキシ基（－Ａｒ－Ｒ－Ｏ－；Ａｒはアリーレン基、Ｒはアルカンジイル基）が挙げられる。Ｑは、エステル結合またはアリーレンオキシ基が好ましい。

In formula (2), R ⁶ is a hydrogen atom, a methyl group or a trifluoromethyl group, preferably a methyl group. R ⁷ is an alkanediyl group having 1 to 10 carbon atoms. Q is an oxygen atom, an ester bond, an amide bond, an arylene group, an alcandiyl group having 1 to 10 carbon atoms, or these groups (that is, an oxygen atom, an ester bond, an amide bond, an arylene group, an alkane having 1 to 10 carbon atoms). It is a combination of (zyl groups). The specific examples of each group in R ⁷ and Q are the same as the specific examples of each group described in R ⁴ , R ⁵ and Q in the formula (1). Examples of the combination of the groups include an aryleneoxy group (-Ar-O-; Ar is an arylene group) and an aralkyleneoxy group (-Ar-RO-; Ar is an arylene group and R is an alkanediyl group). Can be mentioned. Q is preferably an ester bond or an aryleneoxy group.

A is a cyclic ether-containing group or a cyclic carbonate-containing group. It is preferable that R ⁷ in the formula (2) is bound to the cyclic ether or cyclic carbonate in A.
Examples of the cyclic ether-containing group include a 3- to 6-membered ring (cyclic ether ring) -containing group, and specific examples thereof include an epoxidized cyclohexyl group such as an oxylanyl group, an oxetanyl group, and a 3,4-epoxycyclohexyl group. Be done. The cyclic ether ring may have 1 or 2 or more substituents bonded to the ring carbon, and the substituents include, for example, an alkyl group having 1 to 10 carbon atoms such as a methyl group and an ethyl group, and a carbon such as a phenyl group. Examples thereof include an aryl group having a number of 6 to 10.

Examples of the cyclic carbonate-containing group include a cyclic carbonate group having 3 to 6 ring carbon atoms, and examples thereof include an ethylene carbonate group, a propylene carbonate group, and a butylene carbonate group. The cyclic carbonate group may have 1 or 2 or more substituents bonded to the ring carbon, and the substituents include, for example, an alkyl group having 1 to 10 carbon atoms such as a methyl group and an ethyl group, and a carbon such as a phenyl group. Examples thereof include an aryl group having a number of 6 to 10.

Examples of the polymerizable compound that leads to the structural unit (II) include glycidyl (meth) acrylic acid, 2-methylglycidyl (meth) acrylic acid, 3,4-epoxybutyl (meth) acrylic acid, and 6 (meth) acrylic acid. , 7-Epoxyheptyl, (meth) acrylic acid 3,4-epoxycyclohexylmethyl, o, m or p-vinylphenylglycidyl ether, o, m or p-isopropenylphenylglycidyl ether, o, m or p-vinyl Benzyl glycidyl ether, o, m or p-isopropenyl benzyl glycidyl ether; 3-((meth) acryloyloxymethyl) oxetane, 3-((meth) acryloyloxymethyl) -2-methyloxetane, 3-((meth)) Acryloyloxymethyl) -3-ethyloxetane, 3-((meth) acryloyloxymethyl) -2-phenyloxetane, 3- (2- (meth) acryloyloxyethyl) oxetane, 3- (2- (meth) acryloyloxy Ethyl) -2-ethyloxetane, 3- (2- (meth) acryloyloxyethyl) -3-ethyloxetane, 3- (2- (meth) acryloyloxyethyl) -2-phenyloxetane, 3- (3-( Examples include acryloyloxypropyl) oxetane, 3- (p-vinylphenyloxymethyl) -3-methyloxetane, and 3- (p-isopropenylphenyloxymethyl) -3-methyloxetane.
The following examples are particularly preferable for the structural unit (II).

<Structural unit (III)>
The polymer (A) may further have a structural unit (III) other than the structural unit (I) and the structural unit (II). The polymer (A) can have one or more structural units (III).

Examples of the polymerizable compound that leads to the structural unit (III) include, for example.
(Meta) Acrylic Acid Chain Alkyl Esters, For example, (meth) Acrylic Acid Methyl, (Meta) Acrylic Acid Ethyl, (Meta) Acrylic Acid n-Butyl, (Meta) Acrylic Acid sec-Butyl, (Meta) Acrylic Acid t -Butyl, 2-ethylhexyl (meth) acrylate, isodecyl (meth) acrylate, n-lauryl (meth) acrylate, tridecyl (meth) acrylate, n-stearyl (meth) acrylate;
Esters containing (meth) acrylic acid alicyclic, for example, (meth) acrylic acid cyclohexyl, (meth) acrylic acid 2-methylcyclohexyl, (meth) acrylic acid tricyclo [5.2.1.0 ^2,6 ] decane-8. -Il, tricyclo (meth) acrylate [5.2.1.0 ^2,6 ] decane-8-yloxyethyl, isobornyl (meth) acrylate;
(Meta) Acrylic acid aromatic ring-containing ester, for example, (meth) acrylic acid aryl ester such as (meth) acrylic acid phenyl, (meth) acrylic acid aralkyl ester such as (meth) benzyl acrylic acid;
Unsaturated aromatic compounds such as styrene compounds such as styrene, α-methylstyrene, o-methylstyrene, m-methylstyrene, p-methylstyrene, p-methoxystyrene (excluding hydroxylated styrene compounds);
Carboxylic acid-based unsaturated compounds, for example, unsaturated monocarboxylic acids such as (meth) acrylic acid and crotonic acid; unsaturated dicarboxylic acids such as maleic acid, fumaric acid, citraconic acid, mesaconic acid and itaconic acid; the unsaturated dicarboxylic acid. Acid anhydrides; mono-[(meth) acryloyloxyalkyl] esters of polyvalent carboxylic acids such as monosuccinic acid mono [2- (meth) acryloyloxyethyl], mono-phthalic acid [2- (meth) acryloyloxyethyl];
Hydroxyl-containing unsaturated compounds, such as 2-hydroxyethyl (meth) acrylate, 3-hydroxypropyl (meth) acrylate, 4-hydroxybutyl (meth) acrylate, 5-hydroxypentyl (meth) acrylate, (meth). (Meta) Acrylic acid ester having an alcoholic hydroxyl group such as (meth) hydroxyhydroxyhexyl (meth) acrylate; 2-hydroxyphenyl (meth) acrylate, 4-hydroxyphenyl (meth) acrylate (Meta) Acrylic acid ester having a phenolic hydroxyl group such as (meth) hydroxyaryl acrylate; o-hydroxystyrene, p-hydroxystyrene, α-methyl-p-hydroxystyrene, o-hydroxymethylstyrene, p. -Hydroxymethyl styrene compounds such as hydroxymethyl styrene;
Maleimide compounds such as N-phenylmaleimide, N-cyclohexylmaleimide, N-benzylmaleimide, N- (4-hydroxyphenyl) maleimide, N- (4-hydroxybenzyl) maleimide, N-succinimidyl-3-maleimidebenzoate, N. -Succinimidyl-4-maleimidebutyrate, N-succinimidyl-6-maleimide caproate, N-succinimidyl-3-maleimidepropionate, N- (9-acridinyl) maleimide;
In addition, conjugated diene such as 1,3-butadiene, isoprene, 2,3-dimethyl-1,3-butadiene, unsaturated dicarboxylic acid diester such as diethyl maleate, diethyl fumarate, diethyl itaconic acid, (meth) acrylonitrile, (Meta) acrylamide, vinyl chloride, vinylidene chloride, vinyl acetate;
Can be mentioned.

Among these, from the viewpoint of storage stability and patterning property of the resin composition, the polymerizable compound that leads to the structural unit (III) is a (meth) acrylic acid chain alkyl ester, a (meth) acrylic acid alicyclic-containing ester, and the like. At least one selected from the (meth) acrylic acid aromatic ring-containing ester and the unsaturated aromatic compound is preferable, and the (meth) acrylic acid aromatic ring-containing ester is more preferable.

<Structure, physical properties, etc. of polymer (A)>
When the polymer (A) is a copolymer, the mode of arrangement of each structural unit of the repetition is not particularly limited, and any of a random copolymer, an alternate copolymer, a block copolymer and a graft copolymer is used. It may be.

The content ratio of the structural unit (I) in the polymer (A) is preferably 10 mol% or more, more preferably 20 mol% or more, and preferably 70 mol% or less, out of 100 mol% of the total structural unit. It is more preferably 50 mol% or less, specifically, preferably 10 to 70 mol%, and more preferably 20 to 50 mol%. When it is at least the above lower limit value, cells can be satisfactorily aggregated to form a cell mass during cell culture, and it is preferable in terms of developability, and when it is at least the above upper limit value, in terms of developability and resolution. preferable.

The content ratio of the structural unit (II) in the polymer (A) is preferably 10 mol% or more, more preferably 20 mol% or more, and preferably 70 mol% or less, out of 100 mol% of the total structural unit. It is more preferably 40 mol% or less, specifically, preferably 10 to 70 mol%, and more preferably 20 to 40 mol%. When it is at least the above lower limit value, it is preferable in terms of pattern formation and resolvability, and when it is at least the above upper limit value, it is preferable in terms of storage stability and developability of the resin composition.

The content ratio of the structural unit (III) in the polymer (A) is not particularly limited as long as the content ratios of the structural units (I) and (II) are within the above range. For example, the content ratio of the structural unit (III) in 100 mol% of the total structural unit is preferably 80 mol% or less, more preferably 60 mol% or less, and preferably 10 mol% or more. Is preferably 10 to 60 mol%.

The structure of the polymer (A) and the content of structural units can be calculated from the amount of each polymerizable compound charged at the time of synthesizing the polymer (A), but ¹ H-NMR and ¹³ C-NMR. It can also be measured by analysis. For example, using the apparatus name "ECP-400P" (manufactured by JEOL Ltd.), a deuterated solvent having the highest solubility of the polymer is selected from deuterated chloroform, deuterated methanol and heavy water.

The weight average molecular weight (Mw) of the polymer (A) is usually 1000 to 10000, preferably 2000 to 8000, and more preferably 2000 to 6000. In such an embodiment, it is possible to form a pattern film having high resolution and a large film thickness.

The ratio of the weight average molecular weight (Mw) to the number average molecular weight (Mn) of the polymer (A), that is, the molecular weight distribution (Mw / Mn) is usually 1.5 to 4.5, preferably 1.5 to 3. It is 0.0, more preferably 1.5 to 2.5. Such an embodiment is preferable in terms of resolution.

Mw and Mn are measured by gel permeation chromatography (GPC) and are polystyrene-equivalent values. Details of the measurement conditions of GPC are described in Examples.
The total content of the polymer (A) and the radiation-sensitive compound (B) in the resin composition of the present invention is preferably 1% by mass or more, more preferably 2% by mass or more, still more preferably 2% by mass or more, based on 100% by mass of the solid content. Is 5% by mass or more, and in one embodiment, it can be, for example, 10% by mass or more or 20% by mass or more. In such an embodiment, it is easy to control the coating of the resin composition, and the film thickness of the resin composition can be further increased. The solid content is all components except the solvent.

<Method for synthesizing polymer (A)>
The method for synthesizing the polymer (A) is not particularly limited, and a known method can be adopted. For example, it can be synthesized by subjecting a polymerizable compound that leads to the above-mentioned structural unit to a polymerization reaction in the presence of a polymerization initiator in a solvent.

Examples of the solvent include the solvents described in the column of <Method for preparing composition> described later. The amount of solvent is not particularly limited. As the polymerization initiator, one generally known as a radical polymerization initiator can be used. Examples of the radical polymerization initiator include 2,2'-azobisisobutyronitrile, 2,2'-azobis- (2,4-dimethylvaleronitrile), and 2,2'-azobis- (4-methoxy-). Azo compounds such as 2,4-dimethylvaleronitrile); benzoyl peroxide, lauroyl peroxide, diisopropylperoxydicarbonate, t-butylperoxy-2-ethylhexanoate, t-butylperoxypivalate, t-butylperoxydiisob Tyrate, t-butylperoxyneodecanoate, succinic peroxide peroxide, glutalperoxide, succinylperoxyglutarate, t-butylperoxymalate, t-butylperoxypivalate, di-2-ethoxyethylperoxycarbonate, 3-hydroxy Examples thereof include organic peroxides such as -1,1-dimethylbutylperoxypivalate. The polymerization initiator is usually used in an amount of 1 to 10 parts by mass with respect to 100 parts by mass of the polymerizable compound.

<Radiation-sensitive compound (B)>
The radiation-sensitive compound (B) is at least one selected from a radiation-sensitive acid generator and a radiation-sensitive base generator. By using an acid generator or a base generator, low temperature curing of the radiation-sensitive resin composition can be promoted. In the case of low temperature curing, adverse effects such as thermal decomposition on the betaine structure of the polymer (A) can be suppressed. Further, as described above, even if these are used, since the polymer (A) having the structural unit (I) is present, adverse effects on the cells such as deterioration of cell aggregation are suppressed.

Examples of the radiation-sensitive acid generator include compounds that generate an acid by exposure to radiation such as visible light, ultraviolet rays, far ultraviolet rays, electron beams, and X-rays. Specific examples of the radiation-sensitive acid generator include an iodonium salt-based, sulfonium salt-based, tetrahydrothiophenium salt-based, imide sulfonate-based or oxime sulfonate-based radiation-sensitive acid generator, and a quinonediazide compound. .. The radiation-sensitive acid generator may be used alone or in combination of two or more.

Examples of the iodonium salt-based radiation-sensitive acid generator include diphenyliodonium trifluoromethanesulfonate, diphenyliodonium pyrenesulfonate, diphenyliodonium dodecylbenzenesulfonate, diphenyliodonium nonafluoro n-butanesulfonate, and bis (4-t-butylphenyl) iodonium. Trifluoromethanesulfonate, bis (4-t-butylphenyl) iodonium dodecylbenzenesulfonate, bis (4-t-butylphenyl) iodoniumnaphthalene sulfonate, bis (4-t-butylphenyl) iodonium hexafluoroantimonate, bis (4-t-butylphenyl) Examples thereof include t-butylphenyl) iodonium nonafluoro n-butanesulfonate and diphenyliodonium hexafluoroantimonate.

Examples of the sulfonium salt-based radiation-sensitive acid generator include triphenylsulfonium trifluoromethanesulfonate, triphenylsulfonium hexafluoroantimonate, triphenylsulfoniumnaphthalenesulfonate, triphenylsulfonium nonafluoro-n-butanesulfonate, and (hydroxyphenyl). Benzene Methyl Sulfonium Toluene Sulfonium, Cyclohexyl Methyl (2-oxocyclohexyl) Sulfonium Trifluoromethane Sulfonium, Dicyclohexyl (2-oxocyclohexyl) Sulfonium Trifluoromethanesulfonate, Dimethyl (2-oxocyclohexyl) Sulfonium Trifluoromethanesulfonate, Triphenylsulfonium Campersulfonate, ( 4-Hydroxyphenyl) benzylmethyl sulfonium toluene sulfonate, 1-naphthyl dimethyl sulfonium trifluoromethane sulfonate, 1-naphthyl dimethyl sulfonium trifluoromethane sulfonate "1-naphthyl dimethyl" in 1-naphthyl diethyl, 4-cyano-1-naphthyl dimethyl, 4-Nitro-1-naphthyldimethyl, 4-methyl-1-naphthyldimethyl, 4-cyano-1-naphthyl-diethyl, 4-nitro-1-naphthyldiethyl, 4-methyl-1-naphthyldiethyl, 4-hydroxy- Examples include compounds that replace 1-naphthyldimethyl.

Examples of the tetrahydrothiophenium salt-based radiation-sensitive acid generator include 4-hydroxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-methoxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, and 4-ethoxy. -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-methoxymethoxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-ethoxymethoxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4- (1-methoxy) Ethoxy) -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4- (2-methoxyethoxy) -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-methoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate , 4-ethoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-n-propoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-i-propoxycarbonyloxy-1-naphthyltetrahydrothio Phenium trifluoromethanesulfonate, 4-n-butoxycarbonyloxy-1-naphthyltetrahydrothiopheniumtrifluoromethanesulfonate, 4-t-butoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4- (2-tetrahydrofla) Nyloxy) -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4- (2-tetrahydropyranyloxy) -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-benzyloxy-1-naphthyltetrahydrothiophenium trifluo Examples thereof include lomethanesulfonate, 1- (naphthylacetmethyl) tetrahydrothiophenium trifluoromethanesulfonate, 1- (4,7-dibutoxy-1-naphthyl) tetrahydrothiophenium trifluoromethanesulfonate.

Examples of the imide sulfonate-based radiation-sensitive acid generator include trifluoromethylsulfonyloxybicyclo [2.2.1] hept-5-endicarboxyimide, succinimide trifluoromethylsulfonate, phthalimidetrifluoromethylsulfonate, and N-hydroxy. Examples thereof include naphthalimide methanesulfonate, N-hydroxynaphthalimide trifluoromethanesulfonate, and N-hydroxy-5-norbornen-2,3-dicarboxyimide propanesulfonate.

Examples of the oxime sulfonate-based radiation-sensitive acid generator include (5-propylsulfonyloxyimino-5H-thiophen-2-iriden)-(2-methylphenyl) acetonitrile and (5-octylsulfonyloxyimino-5H-thiophene). -2-Ilidene)-(2-Methylphenyl) acetonitrile, (Camfersulfonyloxyimino-5H-thiophene-2-iriden)-(2-Methylphenyl) acetonitrile, (5-p-toluenesulfonyloxyimine-5H-thiophene) -2-Ilidene)-(2-Methylphenyl) acetonitrile, (5-octylsulfonyloxyimino)-(4-methoxyphenyl) acetonitrile can be mentioned.

Examples of the quinone diazide compound include 1,2-naphthoquinone diazide sulfonic acid ester of trihydroxybenzophenone, 1,2-naphthoquinone diazido sulfonic acid ester of tetrahydroxybenzophenone, 1,2-naphthoquinone diazido sulfonic acid ester of pentahydroxybenzophenone, and hexa. Examples thereof include 1,2-naphthoquinone diazide sulfonic acid ester of hydroxybenzophenone and 1,2-naphthoquinone diazido sulfonic acid ester of (polyhydroxyphenyl) alkane.

Examples of the radiation-sensitive base generator include compounds that generate a base by exposure to radiation such as visible light, ultraviolet rays, far ultraviolet rays, electron beams, and X-rays. Specific examples of the radiation-sensitive base generator include 4- (methylthiobenzoyl) -1-methyl-1-morpholinoetan, (4-morpholinobenzoyl) -1-benzyl-1-dimethylaminopropane, and 2-benzyl. Heterocyclic group-containing sensitive radiation such as -2-dimethylamino-1- (4-morpholinophenyl) -butanone, N- (2-nitrobenzyloxycarbonyl) pyrrolidine, 1- (anthraquinone-2-yl) ethylimidazole carboxylate, etc. Sexual base generators; 2-nitrobenzylcyclohexylcarbamate, [[(2,6-dinitrobenzyl) oxy] carbonyl] cyclohexylamine, bis [[(2-nitrobenzyl) oxy] carbonyl] hexane-1,6-diamine, Triphenylmethanol, o-carbamoyl hydroxylamide, o-carbamoyl oxime, hexaammine cobalt (III) tris (triphenyl methyl borate), 1,2-dicyclohexyl-4,4,5,5-tetramethylbiguanidium n- Examples include butyltriphenylborate. The radiation-sensitive base generator may be used alone or in combination of two or more.

The content of the radiation-sensitive compound (B) in the resin composition of the present invention is preferably more than 0 parts by mass and 10 parts by mass or less, and more than 0 parts by mass with respect to 100 parts by mass of the polymer (A). More preferably, it is 5 parts by mass or less. The content of the compound (B) is preferably 0.5 parts by mass or more, more preferably 1 part by mass or more, based on 100 parts by mass of the polymer (A). In such an embodiment, the radiation sensitivity of the composition can be further increased.

<Other additives>
In addition, the resin composition of the present invention includes a compound capable of reacting with the polymer (A), an adhesion aid, a surfactant, an acid or a base diffusion control agent, crosslinked fine particles, a leveling agent, a sensitizer, an inorganic filler, and the like. Various additives such as a quencher can be contained within a range that does not impair the object and properties of the present invention.

Examples of the compound capable of reacting with the polymer (A) include crosslinkable compounds such as an epoxy-based cross-linking agent and an oxetane-based cross-linking agent when the polymer (A) has a carboxy group; the polymer (A). ) Has a structural unit (II), a polyfunctional carboxylic acid such as trimellitic acid can be mentioned.

<Preparation method of composition>
The resin composition of the present invention can be prepared by uniformly mixing each component. In addition, a solvent may be used in the preparation in order to improve the handleability of the composition and to adjust the viscosity and storage stability. Further, in order to remove foreign substances, each component may be uniformly mixed, and then the obtained mixture may be filtered with a filter or the like.

As the solvent, for example,
Alcohol-based solvents such as methanol, ethanol, isopropanol, 1-butanol, 2-butanol, isobutyl alcohol, t-butyl alcohol, 1-hexanol, 1-octanol, 1-nonanol, 1-dodecanol, diacetone alcohol and the like alkyl Alcohol; Aromatic alcohol such as benzyl alcohol ;;
Ether-based solvents such as ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl ether, ethylene glycol monobutyl ether and other ethylene glycol monoalkyl ethers; propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl Propropylene glycol monoalkyl ethers such as ethers and propylene glycol monobutyl ethers; Diethylene glycol monoalkyl ethers such as diethylene glycol monomethyl ethers and diethylene glycol monoethyl ethers; Diethylene glycol dialkyl ethers such as diethylene glycol dimethyl ethers and diethylene glycol ethylmethyl ethers; Dipropylene glycol monomethyl ethers and dipropylenes. Dipropylene glycol monoalkyl ethers such as glycol monoethyl ether, dipropylene glycol monopropyl ether, dipropylene glycol monobutyl ether ;;
Ester-based solvents such as carboxylic acid esters such as ethyl acetate, i-propyl acetate, n-butyl acetate, amyl acetate, ethyl lactate, methyl 3-methoxypropionate, ethyl 3-ethoxypropionate; propylene glycol diacetate and the like. Polyhydric alcohol carboxylate solvent; Polyhydric alcohol partial ether carboxylate solvent such as propylene glycol monomethyl ether acetate and propylene glycol monoethyl ether acetate ;;
Ketone solvents such as acetone, methyl ethyl ketone, diethyl ketone, methyl isobutyl ketone, methyl amyl ketone, diisobutyl ketone, cyclopentanone, cyclohexanone, cycloheptanone ;;
Can be mentioned.

The solvent is preferably a solvent having an octanol / water partition coefficient of 0 or less. The lower limit of the coefficient is not particularly limited, but is, for example, −0.5. The solvent is preferable from the viewpoint of storage stability of the resin composition. Examples of the solvent having an octanol / water partition coefficient of 0 or less include ethyl lactate, acetonitrile, and propylene glycol monomethyl ether. The octanol / water partition coefficient is a value measured by "Measurement of partition coefficient (1-octanol / water) -flask shaking method" of Japanese Industrial Standards (JIS Z 7260-107 (2000)).

The solvent may be used alone or in combination of two or more.
The resin composition of the present invention may contain a solvent in the range of preferably 30 to 90% by mass, more preferably 40 to 88% by mass, still more preferably 45 to 85% by mass.

The resin composition of the present invention has a small increase in viscosity during storage and is excellent in storage stability.
The resin composition of the present invention is preferably a negative type, and by using the resin composition, the size of the non-existent portion of the film and the thickness of the pattern film are independently, usually 0.1 μm or more. A 1 mm pattern film can be formed. Such a pattern membrane is preferable from the viewpoint of cell culture, such as being able to form a cell mass in which cells are three-dimensionally aggregated, and also has a small amount of development residue and can contribute to cell orientation culture.

Further, by using the pattern membrane of the present invention, it is possible to satisfactorily obtain cultured cells arranged two-dimensionally, for example, formed in a row. As an advantage expected from the two-dimensional arrangement, for example, it can be expected that biomimetics (liver shape, etc.) can be performed more accurately.

[Manufacturing method of pattern film]
The method for producing a pattern film of the present invention includes a step of forming a resin film of the radiation-sensitive resin composition of the present invention on a substrate (1), a step of exposing the resin film (2), and the exposed resin. It has a step (3) of developing a film with a developing solution.

In the present invention, the steps (1) to (3) can be carried out at a temperature of preferably 200 ° C. or lower, more preferably 150 ° C. or lower, still more preferably 120 ° C. or lower. This is because the use of the radiation-sensitive compound (B) can promote low-temperature curing of the radiation-sensitive resin composition. In the case of low temperature curing, adverse effects such as thermal decomposition on the betaine structure of the polymer (A) can be suppressed.

<Process (1)>
In the step (1), for example, the resin composition of the present invention is applied and dried on a substrate to form a resin film. The drying conditions are, for example, usually 80 to 150 ° C., preferably 80 to 120 ° C., usually 1 to 5 minutes using an oven or a hot plate. The thickness of the resin film is usually 0.1 μm to 1 mm, preferably 0.1 to 50 μm, and more preferably 0.1 to 10 μm. If the film thickness is insufficient, a resin film may be formed by applying twice.

Examples of the method for applying the composition include a dipping method, a spray method, a bar coating method, a roll coating method, a spin coating method, a curtain coating method, a gravure printing method, a silk screen method, and an inkjet method.

As the substrate, for example, a resin made of at least one resin selected from polystyrene, polycarbonate, polyacetal, polyethylene, polypropylene, polyphenylene sulfide, polyether sulfone, polyethylene terephthalate, polyethylene naphthalate, polyacrylate, polymethacrylate and cellulose. Substrate; Substrate made of a material such as glass, ceramic, stainless steel; Silicon wafer substrate can be mentioned. It may be a resin substrate made of a biodegradable polymer such as polylactic acid, polyglycolic acid, or polycaprolactan.

As the substrate, a substrate for cell culture can be used. As the substrate, a known substrate used for cell culture can be used. For example, it is outside from the honeycomb structure film described in JP-A-2002-335949 and the three-dimensional culture plate manufactured by ORGANOGENIX Co., Ltd. An example is a bottom substrate from which the frame has been removed.

The surface of the substrate may have a fine structure having irregularities. By providing such a microstructure, the adhesiveness of the cell tissue to the surface can be adjusted. For example, a fine mesh structure can be mentioned, and examples of the mesh structure include polygonal shapes such as a circular shape, an elliptical shape, a square shape, and a honeycomb shape.

As the substrate, it is preferable to use a substrate having an appropriate cell adhesion surface from the viewpoint of retaining cells and forming a cell tissue. The cell-adhesive surface is, for example, a surface into which a functional group having a charge such as a carboxy group or an amino group is introduced, a surface into which a cell-adhesive peptide such as an arginine / glycine / aspartic acid sequence is introduced, or a cell-adhesive surface. It is a surface on which the polymer to be possessed is fixed.

The charged functional group can be introduced by treating the surface of the substrate with plasma or the like. As described above, the substrate may be a plasma processing substrate. Examples of the cell-adhesive polymer include charged synthetic polymers such as polyacrylic acid, polyvinyl sulfate, polystyrene sulfonic acid, and polyallylamine, chondroitin sulfate, dermatane sulfate, dextran sulfate, keratane sulfate, and heparan sulfate. Examples thereof include charged polysaccharides such as hyaluronic acid and chitin, cell adhesion proteins such as collagen, gelatin, fibronectin, hydronectin and laminin, and synthetic polymers on which cell adhesion proteins and cell adhesion peptides are immobilized.

<Process (2)>
In step (2), at least a part of the resin film is selectively exposed via a desired pattern mask, for example, using a contact aligner, a stepper or a scanner. Examples of the exposure light include radiation such as visible light, ultraviolet rays, far ultraviolet rays, electron beams, and X-rays, and light having a wavelength of 200 to 500 nm (eg, i-ray (365 nm)) is preferable. The exposure amount varies depending on the type and content of each component in the radiation-sensitive resin composition, the thickness of the resin film, and the like, but the exposure amount is usually 100 to 10,000 mJ / cm ² .

Further, in order to further promote the crosslinking reaction, it is preferable to perform heat treatment after exposure. The heating conditions vary depending on the type and content of each component in the radiation-sensitive resin composition, the thickness of the resin film, and the like, but are usually 80 to 200 ° C, preferably 80 to 150 ° C, and more preferably 100. It is usually about 0.5 to 10 minutes, preferably about 1 to 5 minutes at about 120 ° C. Within the temperature range, adverse effects such as thermal decomposition on the betaine structure of the polymer (A) can be suppressed, which is preferable, and it is also preferable from the viewpoint of patterning performance.

<Process (3)>
In the step (3), the exposed resin film is developed with a developing solution, and in the case of a negative type, the unexposed portion is dissolved and removed to form a pattern film having a desired pattern on the substrate. The size of the non-existent portion of the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 750 μm, and more preferably 0.1 to 500 μm. The lower limit of the size of the non-existent portion is preferably 10 μm, more preferably 20 μm. The thickness of the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 50 μm, and more preferably 0.1 to 10 μm.

As the developing solution, for example, water such as ultrapure water or a mixed solvent containing water is preferable. Examples of the solvent other than water constituting the mixed solvent include organic solvents that can be uniformly mixed with water, such as alcohols such as methanol, ethanol, n-butanol, propylene glycol monomethyl ether and ethyl lactate, and ketones such as cyclopentanone. Kind is mentioned. In the mixed solvent, water is usually 20% by mass or more, preferably 30% by mass or more, and more preferably 40% by mass or more. Among these, a water developer containing water is preferable.

Examples of the developing method include a shower developing method, a spray developing method, a dipping developing method, and a paddle developing method. For example, the resin film formed on the substrate is developed with a developing solution at 20 to 40 ° C. under the condition of about 1 to 10 minutes.
As described above, the pattern film can be manufactured.

[Pattern board]
The pattern substrate of the present invention has a substrate and a pattern film on the substrate. The size of the non-existent portion of the pattern film and the thickness of the pattern film are independently 0.1 μm to 1 mm, and the pattern film is the structural unit (I) represented by the above-mentioned formula (1). Contains a polymer having.

The polymer may be the above-mentioned polymer (A), or may be a polymer in which the structural unit (II) preferably contained in the polymer (A) forms a crosslinked structure.
Examples of the substrate include the specific examples described above as the substrate in <Step (1)>.

The size of the non-existent portion of the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 750 μm, and more preferably 0.1 to 500 μm. The lower limit of the size of the non-existent portion is preferably 10 μm, more preferably 20 μm. The thickness of the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 50 μm, and more preferably 0.1 to 10 μm. A pattern membrane having a structure of the lower limit or more is preferable from the viewpoint of cell culture because it can form a cell mass in which cells are three-dimensionally aggregated.

The size of the non-existent portion of the film is, for example, the dimension when the exposed portion of the substrate is viewed in a plan view without the presence of the film containing the polymer having the structural unit (I). That is, it is the dimension of the exposed substrate surface. For example, when the non-existent portion of the film is a concave portion such as a circular shape, an elliptical shape, or a polygonal shape, it is the maximum length of the exposed substrate surface in the concave portion, and the non-existent portion of the film is like a line shape. In the case of a recess, it is the distance between the films (the length of the exposed substrate surface in the direction orthogonal to the line). The same applies to the above-mentioned sizes referred to elsewhere in this specification.
The pattern substrate of the present invention can be manufactured by the above-mentioned method for manufacturing a pattern film.

[Surface modifier or cell adhesion inhibitor]
The surface modifier or cell adhesion inhibitor of the present invention contains the above-mentioned polymer (A), has low cytotoxicity, and exhibits an excellent cell adhesion inhibitory effect. The polymer (A) can be used as it is as a surface modifier or a cell adhesion inhibitor, or can be used as a component contained in the surface modifier or the cell adhesion inhibitor.
The surface to which the surface modifier or cell adhesion inhibitor of the present invention is applied is, for example, the surface of various instruments / devices, and specific examples of the instruments / devices are [apparatus with modified surface or surface. It is also described in the column of [Device].

The content of the polymer (A) in the surface modifier or cell adhesion inhibitor of the present invention is preferably 1% by mass or more, more preferably 10% by mass or more, and more preferably 20% by mass or more in the solid content from the viewpoint of cytotoxicity. More preferably, it is by mass or more.

The surface modifier or cell adhesion inhibitor of the present invention may contain at least one selected from a solvent, a bactericidal agent and a preservative, in addition to the polymer (A). Examples of the solvent include water and the solvent described in the column of <Method for preparing composition>. The content of the solvent is usually 10 to 90% by mass, preferably 20 to 80% by mass, and more preferably 30 to 70% by mass.

Cell adhesion prevention means preventing or suppressing adhesion between cells and various surfaces with which the cells come into contact. The reason why this effect is exerted is not always clear, but the structural unit (I) of the polymer (A) hydrophilizes the wall surface of the instrument and the device to which the agent is applied, and prevents or suppresses cell adhesion. It is presumed that it can be done. In addition to cells, it can also contribute to the suppression of adhesion of biological substances and tissues such as proteins, lipids and nucleic acids.

The surface modifier or cell adhesion inhibitor of the present invention can be widely used in the medical / bio field (clinical examination / diagnostic agent), for example, clinical diagnostic agent, clinical diagnostic apparatus, biochip, cell culture device, and the like. Coating agent for biological materials and materials that come into contact with biological substances (solids, instruments, devices, etc.); Conditioning agent for measuring cells for fully automatic analyzers used for diagnosis such as blood tests; Useful as cell adhesion control agent ..

By coating at least a part of the device or device with the surface modifier or cell adhesion inhibitor of the present invention, the surface is modified so that cells are less likely to die and cells are less likely to adhere when used. Can provide equipment and devices.

[Apparatus or device with modified surface]
The surface-modified device or apparatus of the present invention has the above-mentioned polymer (A) on at least a part of the surface (either the inner wall surface or the outer wall surface), and is, for example, a polymer. It has a cell adhesion prevention film containing (A).

The instruments and devices are preferably those for cell culture and medical use.
The instruments include, for example, cell culture instruments; instruments for collecting or sending biological substances / tissues (collectively referred to as "biological substances, etc.") such as injection needles, catheters, and blood glucose meters; blood. Containers for storing biological substances such as bags and test tubes; biological substances such as microchannel devices, microscope peripherals, microwell plates, assay chips, biochips and measuring cells for fully automatic analyzers. Instruments for analysis; biotreatment instruments such as reaction tanks, transfer tubes, transfer pipes, purification instruments and cell culture plates; in vivo such as implants, bone fixation materials, sutures, anti-adhesion membranes and artificial blood vessels. Instruments for implantation; other examples include drug delivery media such as vesicles, microparticles and nanoparticles, gastrocameras, microfibers, nanofibers and magnetic particles.

Devices include, for example, medical devices such as clinical diagnostic devices, biosensors, cardiac pacemakers and implantable biochips; fermentation units, bioreactors.

The device or device having a modified surface of the present invention can be manufactured by providing a film containing the polymer (A) on at least a part of the surface of the device or device. For example, it can be produced by coating at least a part of the surface of an instrument or device with the surface modifier or cell adhesion inhibitor of the present invention. The portion is preferably a site where the device or device comes into contact with the cell when the device or device is used.

Examples of the coating method include coating, spraying, and vapor deposition. Further, the agent may contain a polymerization initiator and a polymerizable compound, the agent may be coated on an instrument or an apparatus, and then the polymerizable compound may be polymerized with heat or light. The agent can also be cured using a cross-linking agent and a cross-linking monomer.

Examples of the coating conditions include a method in which a surface modifier or an anti-cell adhesion agent is brought into contact with an instrument or device at 50 to 200 ° C. for about 1 to 10 minutes, then washed with water and dried.

[Cell culture equipment]
One aspect of the cell culture device of the present invention has a pattern membrane obtained by the above-mentioned production method on a part of the surface. For example, the cell culture apparatus of the present invention has a substrate for cell culture and a pattern membrane obtained by the production method on a part of the surface of the substrate. The substrate for cell culture has been described above.

Further, one aspect of the cell culture apparatus of the present invention has a substrate and a conventionally known pattern membrane, and has the polymer (A) on at least a part of the surface of the pattern membrane, for example, a polymer. The cell adhesion prevention film containing (A) is provided on at least a part of the surface (particularly the side wall of the pattern membrane) defining the non-existent portion of the pattern membrane (hereinafter, also referred to as “cell culture recess” or simply “recess”). The surface is hydrophilized, making it difficult for cells to adhere to the surface.

The cell culture device has one or more cell culture recesses for holding cells and cell tissues (particularly cell clusters) formed from the cells. The pattern membrane is a septum that defines the recess and can control the size of the cell tissue and thus create a good cell tissue.

The bottom surface of the recess is made of the surface of the substrate, and the side surface of the recess is made of a pattern film. The surface of the substrate preferably has appropriate cell adhesion from the viewpoint of cell retention. The septum preferably has a cell adhesion-preventing property in consideration of the ease of taking out the cell tissue from the recess after culturing. Since the radiation-sensitive resin composition of the present invention contains the cell adhesion-preventing polymer (A), a partition wall having the above-mentioned properties can be formed.

The shape of the concave portion is not particularly limited, and examples thereof include a circular shape, an elliptical shape, a polygonal shape, and a line shape. The area of the bottom surface of the recess is appropriately determined depending on the size and type of cells and cell tissues.

The size of the recesses in the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 750 μm, and more preferably 0.1 to 500 μm. The lower limit of the size of the recess is preferably 10 μm, more preferably 20 μm. The depth of the recess is determined by the thickness of the pattern film, and is usually 0.1 μm to 1 mm, preferably 0.1 to 50 μm, and more preferably 0.1 to 10 μm. Within this range, cells and cell tissues can be well retained in the area partitioned by the septum.

[Micro flow path device]
One aspect of the microchannel device of the present invention has a pattern film obtained by the above-mentioned manufacturing method on a part of the inner surface of the microchannel. For example, the pattern membrane defines the microchannel.

Further, in one aspect of the microchannel device of the present invention, the polymer (A) is contained in at least a part of the inner surface of the microchannel, and for example, a cell adhesion prevention film containing the polymer (A) is formed in the microflow. It is present on at least a part of the road surface, preferably almost the entire surface. The inner surface of the flow path is made hydrophilic, and biological substances and the like are less likely to adhere to the surface.

Examples of the microchannel device include microanalytical devices such as microreaction devices (eg, microreactors and microplants), integrated nucleic acid analysis devices, microelectrophoretic devices, microchromatography devices, etc .; mass spectra, liquid chromatography, etc. Microdevices for preparing analytical samples; physicochemical processing devices used for extraction, membrane separation, dialysis, etc .; environmental analysis chips, clinical analysis chips, gene analysis chips (DNA chips), protein analysis chips (proteome chips), sugar chain chips , Chromatograph chips, cell analysis chips, pharmaceutical screening chips and other microchannel chips. Among these, the microchannel tip is preferable.

The microchannel is a site where a small amount of sample (preferably a liquid sample) flows, and the width and depth of the channel are not particularly limited, but all of them are usually about 0.1 μm to 1 mm, preferably 10 μm or more. It is 800 μm.

[Method for producing cell mass]
The method for producing a cell mass of the present invention includes a step of culturing cells using the above-mentioned cell culture device to form a cell mass. When culturing cells, the cells are cultured in the region partitioned by the partition wall in the cell culture device to form a cell mass. For example, a culture medium containing cells is placed on a cell culture instrument, and the cells seeded on the bottom surface of the recess form a three-dimensionally bound cell mass on the surface of the substrate which is the bottom surface. The cell mass is cultured for a long period of time while being stably held in the region partitioned by the septum in a state of being adhered to the bottom surface of the recess without floating in the culture solution.

The type of animal, organ, or tissue is not particularly limited as long as the cells form bonds with each other. Examples of cells include scaffold-dependent cells. Examples thereof include primary cells collected from liver, pancreas, kidney, nerve, skin, bone marrow and the like derived from animals such as humans, pigs, dogs, rats and mice, and established cells, specifically, HepG2 cells. , HeLa cells, cancer cells such as F9 cells, and normal cells. Specific examples include fibroblasts such as 3T3 cells; stem cells such as ES cells, iPS cells, and mesenchymal stem cells (eg, UE7T-13 cells); renal cells such as HEK293 cells; nerve cells such as NT2 cells; Examples thereof include endothelial cells such as UV♀2 cells and HMEC-1 cells; myocardial cells such as H9c2 cells; epithelial cells such as Caco-2 cells; or cells subjected to genetic manipulation or the like. In addition, floating cells such as blood cells such as leukocytes, erythrocytes, and platelets may be used. The cells may be used alone or in combination of two or more.
As the culture medium, an aqueous solution containing necessary salts and / or nutritional components at an appropriate concentration can be used so that the viability and function of the cells can be maintained.

Hereinafter, the present invention will be specifically described based on examples, but the present invention is not limited to these examples. In the description of the following examples and the like, "parts" means "parts by mass" unless otherwise specified.
The measurement method of each physical property value is shown below.

[Weight average molecular weight (Mw) and number average molecular weight (Mn)]
For the polymer (A), Mw and Mn were measured by gel permeation chromatography (GPC) under the following conditions. The molecular weight distribution (Mw / Mn) was calculated from the obtained Mw and Mn.
Equipment: GPC-101 (manufactured by Showa Denko)
GPC column: GPC-KF-801, GPC-KF-802, GPC-KF-803 and GPC-KF-804 bonded (manufactured by Shimadzu GLC).
Mobile phase: Dimethylformamide Column temperature: 40 ° C
Flow velocity: 1.0 mL / min Sample concentration: 1.0 mass%
Sample injection amount: 100 μL
Detector: Differential refractometer Standard material: Monodisperse polystyrene

[Content ratio of structural units]
The content ratio of each structural unit of the polymer (A) was calculated from the charged amount of each polymerizable compound.

<Synthesis of polymer>
The polymerizable compounds used in the synthesis of the polymers of each Example and Comparative Example are shown below.
[Polymerizable compound giving a structural unit (I) or a comparative structural unit (cI)]
In the synthesis example, the polymerizable compounds (1-1) to (1-4) that lead to the structural units represented by the following formulas (I-1) to (I-4) and (cI-1) to (cI-3) are derived. ), (C1-1) to (c1-3) were used.

Ｍｅはメチル基である。

Me is a methyl group.

[Polymerizable compound giving structural unit (II)]
In the synthesis example, the polymerizable compounds (2-1) to (2-9) that derive the structural units represented by the following formulas (II-1) to (II-9) were used.

[Polymerizable compound giving structural unit (III)]
In the synthesis example, the polymerizable compounds (3-1) to (3-5) that derive the structural units represented by the following formulas (III-1) to (III-5) were used.

[Synthesis Example 1] (Synthesis of polymer (A-1))
A flask equipped with a cooling tube and a stirrer was charged with 1.0 part of 2,2'-azobis (2,4-dimethylvaleronitrile) and 40.0 parts of ethyl lactate. Next, 3.6 parts of the polymerizable compound (1-1) giving the structural unit (I), 2.4 parts of the polymerizable compound (2-1) giving the structural unit (II-1), and the structural unit (III-). Add 4.0 parts of the polymerizable compound (3-3) to give 3), replace with nitrogen, raise the temperature of the solution to 70 ° C while gently stirring, and maintain this temperature for 4 hours for polymerization. Obtained a polymer solution containing the polymer (A-1). The solid content concentration of this polymer solution was 21% by mass, the Mw of the polymer (A-1) was 4600, and the molecular weight distribution (Mw / Mn) was 2.3.

[Synthesis Examples 2-32] (Synthesis of Polymer)
The same procedure as in Synthesis Example 1 was carried out except that the type and amount of the polymerizable compound shown in Table 1 was used, and the polymers (A-2) to (A-28) and the polymers (cA-1) to ( cA-4) was synthesized. The solid content concentration of each of the obtained polymer solutions and the Mw and Mw / Mn of each polymer were about the same as the values of the polymer (A-1).

<Preparation of resin composition (1)>
[Example 1]
To a polymer solution containing 100 parts (solid content) of the polymer (A-1), 5 parts of N-hydroxynaphthalimide trifluoromethanesulfonate (NAI-105, Midori Kagaku) was added to make the solid content concentration 20% by mass. After adding ethyl lactate as a solvent, the resin composition (1) was prepared by filtering with a polymer filter having a pore size of 0.2 μm.

[Examples 2 to 32 and Comparative Examples 1 to 4]
Each resin composition was prepared by the same procedure as in Example 1 except that the polymer shown in Table 1 was used as the polymer (A). However, as the solvent, ethyl lactate in Examples 2 to 28 and Comparative Examples 1 to 4, propylene glycol monomethyl ether (PGME) in Example 29, isopropanol (IPA) in Example 30, and propylene glycol in Example 31. Monomethyl ether acetate (PGMEA) was used respectively. The octanol / water partition coefficients of ethyl lactate, PGME, IPA and PGMEA were −0.18, −0.49, 0.05 and 0.36, respectively.

<Evaluation>
Each of the prepared resin compositions was evaluated according to the following evaluation method. The evaluation results are shown in Table 1.
[Storage stability performance]
The viscosity of the resin composition prepared above at 25 ° C. was measured using an ELD type viscometer manufactured by Tokyo Keiki Co., Ltd. Then, the composition was allowed to stand at 5 ° C., and the viscosity at 25 ° C. was measured every day. The number of days required to increase the viscosity by 5% was determined based on the viscosity immediately after preparation. If the number of days is 7 days or less, it is "1", if it is 8 days or more and 14 days or less, it is "2", if it is 15 days or more and 21 days or less, it is "3", and if it is 22 days or more and 28 days or less. The case was evaluated as "4", and the case of 29 days or more was evaluated as "5". The evaluation results are shown in Table 1. It can be said that the larger the value, the less the thickening and the better the storage stability.

[Patterning performance]
After applying each resin composition on a glass substrate using a spin coater, it was prebaked on a hot plate at 90 ° C. for 2 minutes to form a coating film having a film thickness of 3.0 μm. The obtained coating film was irradiated with ultraviolet rays by a mercury lamp via a pattern mask having a dot pattern having a diameter of 20, 30, 40, 50, 60, 70, 80, 90, 100 μm. The exposure amount at a wavelength of 365 nm was set to 300 mJ / cm ² . Next, after baking on a hot plate at 110 ° C. for Examples 1 to 31 and Comparative Examples 1 to 4 and 180 ° C. for 2 minutes in Example 32, development treatment was performed at 25 ° C. for 120 seconds using ultrapure water. .. At this time, "1" is applied when the pattern contrast is not obtained at all, "2" is applied when the pattern contrast is obtained but the pattern "missing" part is not completely removed to the bottom of the coating film, and the pattern "missing" part is painted. "3" when the minimum pattern size that can be pulled out to the bottom of the film is 60 to 100 μm, "4" when the minimum pattern size that can be pulled out to the bottom of the coating film is 30 to 50 μm, and the pattern "missing" part is painted. The case where the minimum pattern size that penetrates to the bottom of the film is 20 μm was evaluated as “5”. The evaluation results are shown in Table 1. It can be said that the larger the evaluation value is, the smaller the size of the pattern can be formed and the better the patterning performance is.

[Cell aggregation]
In culture of UE7T-13 cells (JCRB1154), which are mesenchymal stem cells (hMSC) derived from human bone marrow, 10% by mass FBS (GE Healthcare), 100 U / mL penicillin, and 100 μg / mL streptomycin (Thermo Fisher Scientific). Dalvecco modified Eagle's medium (Wako Pure Chemical Industries, Ltd.) containing FIC) was used. The cells were maintained by adhering culture using a cell culture flask (Thermo Fisher Scientific) in an environment of 37 ° C., volume fraction of CO ₂₅ %, and air of 95%. When the cells were used in the experiment, the cells in the logarithmic growth phase were exfoliated by TrypLE ^TM Express (Thermo Fisher Scientific), and then the cells were suspended in a medium to prepare a single cell suspension. Similar to [Patterning performance], a glass substrate having a thin film having a thickness of 2 μm formed by 100 hole patterns having a diameter of 100 μm was placed on a cell culture vessel fractionated in a silicon chamber (flexiPERM micro) manufactured by Zalstad. UE7T-13 cells of 2.1 × 10 ⁴ Cells / cm ² were seeded and cultured at 37 ° C. for 4 days in an atmosphere having a volume fraction of CO ₂₅ % and air 95%.

Four days later, the aggregate of cells in the whole pattern was observed in a bright field with an optical microscope (Olympus, IX71). A cell is defined as "○" when cells are present only in the whole part, "△" when most cells are present in the whole part, and "×" when cells are present in the whole part and other parts without distinction. Aggregation was evaluated. “-” Indicates that the hole pattern could not be formed and therefore cannot be evaluated. The evaluation results are shown in Table 1.

As can be seen from the evaluation results in Table 1, the radiation-sensitive resin composition composed of a polymer containing a specific amount of carbobetaine or sulfobetaine had good properties, whereas it had a quaternary ammonium salt. The radiation-sensitive resin composition composed of the polymer had poor cell aggregation performance characteristics.

Claims (11)

The structural unit (I) represented by the formula (1) and
Structural unit containing at least one selected from cyclic ether structure and cyclic carbonate structure (II)
The polymer (A) having
A radiation-sensitive resin composition containing at least one radiation-sensitive compound (B) selected from a radiation-sensitive acid generator and a radiation-sensitive base generator.

[In formula (1), R ¹ is a hydrogen atom, a methyl group or a trifluoromethyl group, and R ² and R ³ are independently alkyl or aryl groups having 1 to 5 carbon atoms, respectively, and R ⁴ And R ⁵ are independently alcandiyl groups having 1 to 10 carbon atoms, and X is a carboanion (-COO ^- group), a sulfoanion (-SO ₃ ^- group), or a phosphate anion, and Q. Is an oxygen atom, an ester bond, an amide bond, an arylene group, an alcandiyl group having 1 to 10 carbon atoms, or a combination thereof. ] The radiation-sensitive resin composition according to claim 1 , wherein the structural unit (II) includes a cyclic ether structure, and the cyclic ether structure is at least one selected from an oxylan structure and an oxetane structure. The radiation-sensitive resin composition according to claim 1 or 2 , wherein the content ratio of the structural unit (I) is 10 to 70 mol% in 100 mol% of the total structural unit of the polymer (A). The radiation-sensitive resin composition according to any one of claims 1 to 3 , which contains the radiation-sensitive compound (B) in a range of 10 parts by mass or less with respect to 100 parts by mass of the polymer (A). The radiation-sensitive resin composition according to any one of claims 1 to 4 , further comprising a solvent having an octanol / water partition coefficient of 0 or less. (1) A step of forming a resin film of the radiation-sensitive resin composition according to any one of claims 1 to 5 on a substrate.
A method for producing a pattern film, comprising (2) a step of exposing the resin film and (3) a step of developing the exposed resin film with a developing solution. The method for producing a pattern film according to claim 6 , wherein the developer used in the step (3) contains water. The method for producing a pattern film according to claim 6 or 7 , wherein the steps (1) to (3) are performed at a temperature of 200 ° C. or lower. A method for producing a cell culture device having a pattern membrane on a part of the surface produced by the production method according to any one of claims 6 to 8 . A method for manufacturing a microchannel device having a pattern film produced by the manufacturing method according to any one of claims 6 to 8 on a part of the inner surface of the microchannel. A method for producing a cell mass, which comprises a step of culturing cells using the cell culture device produced by the production method according to claim 9 to form a cell mass.