JPWO2018168309A1 - Radiation-sensitive resin composition, pattern film and method for producing the same, pattern substrate, cell culture instrument, microchannel device, and method for producing cell mass - Google Patents

Abstract

[Problem] To provide a radiation-sensitive resin composition which can suppress an acid or a base generated during pattern formation from adversely affecting cells. [Solution] 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. In the formula (1), R 1 is a hydrogen atom, a methyl group or a trifluoromethyl group, R 2 and R 3 are each independently an alkyl group or an aryl group having 1 to 5 carbon atoms, and R 4 and R 5 are each Is independently an alkanediyl group having 1 to 10 carbon atoms, X is a carbanion (-COO- group), a sulfonion (-SO3- group) or a phosphate anion, and Q is an oxygen atom, an ester bond. Amide bond, arylene group, alkanediyl group having 1 to 10 carbon atoms, or a combination of these groups.

Description

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

  In the cell culture technology, attention has been paid to culturing a tissue having a function equivalent to that of a tissue in a living body. The tissue body is, for example, a cell mass formed by three-dimensionally assembling cells (for example, see Patent Documents 1 and 2).

International Publication No. 2007/097120 JP 2006-121991 A

  As a cell culture technique, a pattern film is formed using a radiation-sensitive resin composition, and this pattern film is used as a partition, and cells can be cultured in a region defined by the partition. Here, for example, when a radiation-sensitive acid generator or base generator is used for pattern formation in the resin composition, an acid or base generated during pattern formation has an adverse effect on cells, for example, cell aggregation is deteriorated. May have an adverse effect.

  The present invention provides a radiation-sensitive resin composition capable of suppressing an acid or base generated during pattern formation from exerting the above-mentioned adverse effect on cells, a method for producing a pattern film using the composition, and a pattern film. That is the task. Another object of the present invention is to provide a pattern substrate, a cell culture instrument, and a microchannel device having excellent cell adhesion preventing properties. Another object of the present invention is to provide a method for producing a cell mass.

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

  [1] A polymer (A) having the 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) 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, R ² and R ³ are each independently an alkyl group or an aryl group having 1 to 5 carbon atoms, R ⁴ And R ⁵ are each independently an alkanediyl group having 1 to 10 carbon atoms, X is a carbanion (—COO ⁻ group), a sulfonion (—SO ₃ ⁻ group), or a phosphate anion; Is an oxygen atom, an ester bond, an amide bond, an arylene group, an alkanediyl group having 1 to 10 carbon atoms, or a combination of these groups. ]

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

[4] The composition according to any one of [1] to [3], wherein the content of the structural unit (I) is 10 to 70 mol% based on 100 mol% of all the structural units of the polymer (A). Radiation-sensitive resin composition.
[5] The radiation-sensitive composition according to any one of [1] to [4], wherein the radiation-sensitive compound (B) is contained in an amount of 10 parts by mass or less based on 100 parts by mass of the polymer (A). Resin composition.

[6] The radiation-sensitive resin composition according to any one of [1] to [5], further comprising 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, And (3) a method for producing a pattern film, comprising a step of developing the exposed resin film with a developer.

[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 according to [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 of manufacturing a pattern film according to any one of [7] to [9].
[11] A cell culture instrument having the pattern film according to the above [10] on a part of its surface.

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

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

  Further, according to the present invention, it is possible to provide a pattern substrate, a cell culture instrument and a microchannel device which are excellent in cell adhesion prevention properties, and it is 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 (also simply referred to as “resin composition”) of the present invention comprises a polymer (A) described below and at least one selected from a radiation-sensitive acid generator and a radiation-sensitive base generator. And at least one 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 may have two or more types of structural units (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 film or the like for cell culture, an acid or a base generated from an acid generator or a base generator during pattern formation adversely affects cells, such as deterioration of cell aggregation properties. there is a possibility. However, in the present invention, it is considered that the polymer (A) having a betaine structure called the structural unit (I) acts as a quencher for the acid or the base and traps them, so that the adverse effect on the cell is reduced. 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.

Note that in this specification, a betaine structure means an atom having a positive charge and a negative charge at positions not adjacent to each other (R ^{5 in} Formula (1)) and a positive formal charge (Formula (1) N means a structure in which a dissociable hydrogen atom is not bonded.

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 leading to the structural unit (I) is easily available. R ² and R ³ are each independently an alkyl group or an aryl group having 1 to 5 carbon atoms, preferably an alkyl group. R ⁴ and R ⁵ are each independently an alkanediyl group having 1 to 10 carbon atoms.

X is a carbanion (-COO ^- group), a sulfo anion (-SO ₃ ^- group), or a phosphate anion, preferably carbanion or sulfo anion.
Q represents 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, An alkanediyl group or a combination of these groups (that is, an oxygen atom, an ester bond, an amide bond, an arylene group, an alkanediyl group having 1 to 10 carbon atoms), and preferably an ester bond or an amide bond.

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

The alkanediyl group for 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, for example, a methanediyl group, an ethane-1,2-diyl group, and a propane-1,3-diyl group.

  The arylene group for Q is preferably an arylene group having 6 to 10 carbon atoms, for example, a phenylene group, a tolylene group, and a naphthylene group. The alkanediyl group for 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 leads to the structural unit (I) include N- (meth) acryloyloxyethyl-N, N-dimethylammonium-α-N-methylcarboxybetaine, N- (meth) acryloyloxyethyl-N, N-dimethylammonium-α-N-ethylcarboxybetaine, N- (meth) acryloyloxyethyl-N, N-dimethylammonium-α-N-propylsulfobetaine, N- (meth) acryloylaminopropyl-N, N- Dimethylammonium-α-N-propylsulfobetaine.
The following examples are particularly preferred as 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-type material and from the viewpoint of crosslinking reactivity. More preferably, it has. The cyclic ether structure is preferably at least one selected from an oxirane structure and an oxetane structure.
The polymer (A) may 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 the formula (2), R ⁶ is a hydrogen atom, a methyl group or a trifluoromethyl group, and is preferably a methyl group. R ⁷ is an alkanediyl group having 1 to 10 carbon atoms. Q represents an oxygen atom, an ester bond, an amide bond, an arylene group, an alkanediyl group having 1 to 10 carbon atoms, or any of these groups (that is, an oxygen atom, an ester bond, an amide bond, an arylene group, an alkane having 1 to 10 carbon atoms) Diyl group). 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), an aralkyleneoxy group (—Ar—R—O—; Ar is an arylene group, and R is an alkanediyl group). No. Q is preferably an ester bond or an aryleneoxy group.

A is a cyclic ether-containing group or a cyclic carbonate-containing group. R ⁷ in the formula (2) is preferably bonded to the cyclic ether or cyclic carbonate in A.
The cyclic ether-containing group is, for example, a 3- to 6-membered ring (cyclic ether ring) -containing group, and specific examples thereof include an oxiranyl group, an oxetanyl group, and an epoxidized cyclohexyl group such as a 3,4-epoxycyclohexyl group. Can be The cyclic ether ring may have one or more substituents bonded to the ring carbon, and examples of the substituent include a carbon group such as an alkyl group having 1 to 10 carbon atoms such as a methyl group and an ethyl group, and a carbon atom such as a phenyl group. Aryl groups of the number 6 to 10 are exemplified.

  The cyclic carbonate-containing group is, for example, 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 one or more substituents bonded to a ring carbon. Examples of the substituent include a carbon group such as an alkyl group having 1 to 10 carbon atoms such as a methyl group and an ethyl group, and a carbon atom such as a phenyl group. Aryl groups of the number 6 to 10 are exemplified.

Examples of the polymerizable compound that leads to the structural unit (II) include glycidyl (meth) acrylate, 2-methylglycidyl (meth) acrylate, 3,4-epoxybutyl (meth) acrylate, and (meth) acrylic acid 6 , 7-epoxyheptyl, 3,4-epoxycyclohexylmethyl (meth) acrylate, o, m or p-vinylphenyl glycidyl ether, o, m or p-isopropenylphenyl glycidyl 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) a Liloyloxymethyl) -2-phenyloxetane, 3- (2- (meth) acryloyloxyethyl) oxetane, 3- (2- (meth) acryloyloxyethyl) -2-ethyloxetane, 3- (2- (meta) ) Acryloyloxyethyl) -3-ethyloxetane, 3- (2- (meth) acryloyloxyethyl) -2-phenyloxetane, 3- (3- (meth) acryloyloxypropyl) oxetane, 3- (p-vinylphenyl) (Oxymethyl) -3-methyloxetane and 3- (p-isopropenylphenyloxymethyl) -3-methyloxetane.
The following examples are particularly preferred 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,
(Meth) acrylic acid chain alkyl ester, for example, methyl (meth) acrylate, ethyl (meth) acrylate, n-butyl (meth) acrylate, sec-butyl (meth) acrylate, t- (meth) acrylate -Butyl, 2-ethylhexyl (meth) acrylate, isodecyl (meth) acrylate, n-lauryl (meth) acrylate, tridecyl (meth) acrylate, n-stearyl (meth) acrylate;
(Meth) acrylic acid alicyclic ester, for example, cyclohexyl (meth) acrylate, 2-methylcyclohexyl (meth) acrylate, tricyclo [5.2.1.0 ^2,6 ] decane-8 methacrylate -Yl, tricyclo [5.2.1.0 ^2,6 ] decane-8-yloxyethyl (meth) acrylate, isobornyl (meth) acrylate;
(Meth) acrylic acid aromatic ring-containing ester, for example, (meth) acrylic acid aryl ester such as phenyl (meth) acrylate, (meth) acrylic acid aralkyl ester such as benzyl (meth) acrylate;
Unsaturated aromatic compounds, for example, styrene compounds such as styrene, α-methylstyrene, o-methylstyrene, m-methylstyrene, p-methylstyrene, p-methoxystyrene (excluding hydroxylated styrene compounds);
Carboxylic acid 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; Acid anhydrides; mono [(meth) acryloyloxyalkyl] esters of polycarboxylic acids such as mono [2- (meth) acryloyloxyethyl] succinate and mono [2- (meth) acryloyloxyethyl] phthalate;
Hydroxyl-containing unsaturated compounds, for example, 2-hydroxyethyl (meth) acrylate, 3-hydroxypropyl (meth) acrylate, 4-hydroxybutyl (meth) acrylate, 5-hydroxypentyl (meth) acrylate, (meth) ) (Meth) acrylic esters having an alcoholic hydroxyl group, such as hydroxyalkyl (meth) acrylates such as 6-hydroxyhexyl acrylate; 2-hydroxyphenyl (meth) acrylate, 4-hydroxyphenyl (meth) acrylate (Meth) acrylates having a phenolic hydroxyl group such as hydroxyaryl (meth) acrylate; o-hydroxystyrene, p-hydroxystyrene, α-methyl-p-hydroxystyrene, o-hydroxymethylstyrene, p -Such as hydroxymethylstyrene Dorokishi styrene compound;
Maleimide compounds such as N-phenylmaleimide, N-cyclohexylmaleimide, N-benzylmaleimide, N- (4-hydroxyphenyl) maleimide, N- (4-hydroxybenzyl) maleimide, N-succinimidyl-3-maleimidobenzoate, N -Succinimidyl-4-maleimidobutyrate, N-succinimidyl-6-maleimidocaproate, N-succinimidyl-3-maleimidopropionate, N- (9-acridinyl) maleimide;
In addition, conjugated dienes such as 1,3-butadiene, isoprene and 2,3-dimethyl-1,3-butadiene, unsaturated dicarboxylic acid diesters such as diethyl maleate, diethyl fumarate and diethyl itaconate, (meth) acrylonitrile, (Meth) acrylamide, vinyl chloride, vinylidene chloride, vinyl acetate;
Is mentioned.

  Among these, from the viewpoints of storage stability and patterning properties of the resin composition, polymerizable compounds that lead to the structural unit (III) include (meth) acrylic acid chain alkyl esters, (meth) acrylic acid alicyclic ester, At least one selected from (meth) acrylic acid aromatic ring-containing esters and unsaturated aromatic compounds is preferable, and (meth) acrylic acid aromatic ring-containing esters are more preferable.

<Configuration and physical properties of polymer (A)>
When the polymer (A) is a copolymer, the arrangement of the repeating structural units is not particularly limited, and may be any of a random copolymer, an alternating copolymer, a block copolymer, and a graft copolymer. It may be.

  The content of the structural unit (I) in the polymer (A) is preferably at least 10 mol%, more preferably at least 20 mol%, and preferably at most 70 mol%, based on 100 mol% of all the structural units. It is more preferably 50 mol% or less, specifically, preferably 10 to 70 mol%, more preferably 20 to 50 mol%. When the amount is not less than the lower limit, it is preferable in terms of the ability to form a cell mass by satisfactorily assembling cells during cell culture and in terms of developability, and in the case of being not more than the upper limit, in terms of developability and resolution. preferable.

  The content of the structural unit (II) in the polymer (A) is preferably at least 10 mol%, more preferably at least 20 mol%, and preferably at most 70 mol%, based on 100 mol% of all the structural units. It is more preferably at most 40 mol%, specifically, preferably 10 to 70 mol%, more preferably 20 to 40 mol%. It is preferable from the viewpoint of pattern formability and resolution when it is not less than the lower limit, and it is preferable from the viewpoint of storage stability and developability of the resin composition when it is not more than the above upper limit.

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

Although the structure of the polymer (A) and the content of the structural unit can be calculated from the charged amounts of the respective polymerizable compounds at the time of the synthesis of the polymer (A), ¹ H-NMR and ¹³ C-NMR It can also be measured by analysis. For example, using a device name “ECP-400P” (manufactured by JEOL), a heavy solvent having the highest polymer solubility is selected from heavy chloroform, heavy methanol and heavy water.

  The weight average molecular weight (Mw) of the polymer (A) is usually from 1,000 to 10,000, preferably from 2,000 to 8,000, more preferably from 2,000 to 6,000. In such an embodiment, a pattern film having high resolution and a large film thickness can be formed.

  The ratio between the weight average molecular weight (Mw) and 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 And 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 values in terms of polystyrene. 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, and still more preferably 100% by mass of solid content. Is 5% by mass or more, and in one embodiment, may be, for example, 10% by mass or more or 20% by mass or more. In such an embodiment, application control of the resin composition can be easily performed, and the thickness of the resin composition can be further increased. The solid content is all components except the solvent.

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

  Examples of the solvent include the solvents described below in the section <Method for Preparing Composition>. The amount of the solvent is not particularly limited. As the polymerization initiator, those generally known as radical polymerization initiators 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-butylperoxydiisobu Tylate, t-butyl peroxy neodecanoate, succinic peroxide, glutar peroxide, succinyl peroxyglutarate, t-butyl peroxymalate, t-butyl peroxypivalate, di-2-ethoxyethyl peroxycarbonate, 3-hydroxy -1,1-dimethylbutyl Organic peroxides such as oxy pivalate and the like. The polymerization initiator is usually used in an amount of 1 to 10 parts by mass based on 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 when these are used, since the polymer (A) having the structural unit (I) is present, adverse effects on cells such as deterioration of cell aggregation are suppressed.

  Examples of the radiation-sensitive acid generator include compounds that generate an acid upon exposure to radiation such as visible light, ultraviolet light, far ultraviolet light, an electron beam, and X-rays. Specific examples of the radiation-sensitive acid generator include, for example, iodonium salt-based, sulfonium salt-based, tetrahydrothiophenium salt-based, imidosulfonate-based or oxime sulfonate-based radiation-sensitive acid generators, and quinonediazide compounds. . 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 pyrene sulfonate, diphenyliodonium dodecylbenzenesulfonate, diphenyliodonium nonafluoro n-butanesulfonate, and bis (4-t-butylphenyl) iodonium. Trifluoromethanesulfonate, bis (4-t-butylphenyl) iodonium dodecylbenzenesulfonate, bis (4-t-butylphenyl) iodonium naphthalene sulfonate, bis (4-t-butylphenyl) iodonium hexafluoroantimonate, bis (4- t-butylphenyl) iodonium nonafluoro n-butanesulfonate, diphenyliodonium hexafluoroantimonate Door and the like.

  Examples of the sulfonium salt-based radiation-sensitive acid generator include triphenylsulfonium trifluoromethanesulfonate, triphenylsulfonium hexafluoroantimonate, triphenylsulfonium naphthalene sulfonate, triphenylsulfonium nonafluoro-n-butanesulfonate, (hydroxyphenyl) Benzenemethylsulfonium toluenesulfonate, cyclohexylmethyl (2-oxocyclohexyl) sulfonium trifluoromethanesulfonate, dicyclohexyl (2-oxocyclohexyl) sulfonium trifluoromethanesulfonate, dimethyl (2-oxocyclohexyl) sulfonium trifluoromethanesulfonate, triphenylsulfonium camphorsulfonate, ( 4-hydroxy Enyl) benzylmethylsulfonium toluenesulfonate, 1-naphthyldimethylsulfonium trifluoromethanesulfonate, 1-naphthyldimethylsulfonium trifluoromethanesulfonate in which "1-naphthyldimethyl" is 1-naphthyldiethyl, 4-cyano-1-naphthyldimethyl, 4-nitro -1-naphthyldimethyl, 4-methyl-1-naphthyldimethyl, 4-cyano-1-naphthyl-diethyl, 4-nitro-1-naphthyldiethyl, 4-methyl-1-naphthyldiethyl, 4-hydroxy-1-naphthyl Compounds substituted for dimethyl can be mentioned.

  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-naphthyltetrahydrothiophene Trifluoromethanesulfonate, 4-methoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-ethoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-n-propoxycarbonyloxy-1-naphthyl Tetrahydrothiophenium trifluoromethanesulfonate, 4-i-propoxycarbonyloxy-1-naphthyl tetrahydrothiophenium trifluoromethanesulfonate, 4-n-butoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-t- Butoxycarbonyloxy-1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4- (2 Tetrahydrofuranyloxy) -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4- (2-tetrahydropyranyloxy) -1-naphthyltetrahydrothiophenium trifluoromethanesulfonate, 4-benzyloxy-1-naphthyltetrahydrothiophenium Trifluoromethanesulfonate, 1- (naphthyl acetomethyl) tetrahydrothiophenium trifluoromethanesulfonate, 1- (4,7-dibutoxy-1-naphthyl) tetrahydrothiophenium trifluoromethanesulfonate.

  Examples of the imidosulfonate-based radiation-sensitive acid generator include, for example, trifluoromethylsulfonyloxybicyclo [2.2.1] hept-5-enedicarboximide, succinimidetrifluoromethylsulfonate, phthalimidotrifluoromethylsulfonate, N-hydroxy Naphthalimide methanesulfonate, N-hydroxynaphthalimidotrifluoromethanesulfonate, N-hydroxy-5-norbornene-2,3-dicarboximidepropanesulfonate are exemplified.

  Examples of the oxime sulfonate-based radiation-sensitive acid generator include (5-propylsulfonyloxyimino-5H-thiophen-2-ylidene)-(2-methylphenyl) acetonitrile and (5-octylsulfonyloxyimino-5H-thiophene). -2-ylidene)-(2-methylphenyl) acetonitrile, (camphorsulfonyloxyimino-5H-thiophen-2-ylidene)-(2-methylphenyl) acetonitrile, (5-p-toluenesulfonyloxyimino-5H-thiophene) -2-ylidene)-(2-methylphenyl) acetonitrile and (5-octylsulfonyloxyimino)-(4-methoxyphenyl) acetonitrile.

  Examples of the quinonediazide compound include 1,2-naphthoquinonediazidesulfonic acid ester of trihydroxybenzophenone, 1,2-naphthoquinonediazidesulfonic acid ester of tetrahydroxybenzophenone, 1,2-naphthoquinonediazidesulfonic acid ester of pentahydroxybenzophenone, hexa Examples thereof include 1,2-naphthoquinonediazidosulfonic acid ester of hydroxybenzophenone and 1,2-naphthoquinonediazidosulfonic acid ester of (polyhydroxyphenyl) alkane.

  Examples of the radiation-sensitive base generator include compounds that generate a base upon exposure to radiation such as visible light, ultraviolet light, far ultraviolet light, an electron beam, and X-rays. Specific examples of the radiation-sensitive base generator include, for example, 4- (methylthiobenzoyl) -1-methyl-1-morpholinoethane, (4-morpholinobenzoyl) -1-benzyl-1-dimethylaminopropane, 2-benzyl Heterocyclic group-containing radiation-sensitive radiation such as -2-dimethylamino-1- (4-morpholinophenyl) -butanone, N- (2-nitrobenzyloxycarbonyl) pyrrolidine, 1- (anthraquinone-2-yl) ethylimidazole carboxylate. Anionic base generator; 2-nitrobenzylcyclohexyl carbamate, [[(2,6-dinitrobenzyl) oxy] carbonyl] cyclohexylamine, bis [[(2-nitrobenzyl) oxy] carbonyl] hexane-1,6-diamine, Triphenyl methanol, o-carbamoyl hydroxyl Imide, o-carbamoyl oxime, hexaamminecobalt (III) tris (triphenylmethyl borate), 1,2-dicyclohexyl-4,4,5,5-tetramethylbiguanidium n-butyl triphenyl borate. . 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, more than 0 parts by mass, based on 100 parts by mass of the polymer (A). 5 parts by mass or less is more preferable. The content of the compound (B) is preferably at least 0.5 part by mass, more preferably at least 1 part by mass, 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>
The resin composition of the present invention may further include a compound capable of reacting with the polymer (A), an adhesion aid, a surfactant, an acid or base diffusion controlling agent, crosslinked fine particles, a leveling agent, a sensitizer, an inorganic filler, 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, when the polymer (A) has a carboxy group, a crosslinkable compound such as an epoxy-based crosslinking agent and an oxetane-based crosslinking agent; ) Has a structural unit (II), and includes a polyfunctional carboxylic acid such as trimellitic acid.

<Method for preparing composition>
The resin composition of the present invention can be prepared by uniformly mixing the components. 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, after the respective components are uniformly mixed, the resulting mixture may be filtered with a filter or the like.

As the solvent, for example,
Alcohol solvents such as methanol, ethanol, isopropanol, 1-butanol, 2-butanol, isobutyl alcohol, t-butyl alcohol, 1-hexanol, 1-octanol, 1-nonanol, 1-dodecanol, alkyls such as diacetone alcohol Alcohols; aromatic alcohols such as benzyl alcohol;
Ether solvents such as ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl ether, ethylene glycol monoalkyl ether such as ethylene glycol monobutyl ether; propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl Propylene glycol monoalkyl ethers such as ether and propylene glycol monobutyl ether; diethylene glycol monoalkyl ethers such as diethylene glycol monomethyl ether and diethylene glycol monoethyl ether; diethylene glycol dialkyl ethers such as diethylene glycol dimethyl ether and diethylene glycol ethyl methyl ether Le; dipropylene glycol monomethyl ether, dipropylene glycol monoethyl ether, dipropylene glycol monopropyl ether, dipropylene glycol monoalkyl ethers and dipropylene glycol monobutyl ether ;;
Ester solvents, for example, carboxylic acid esters such as ethyl acetate, i-propyl acetate, n-butyl acetate, amyl acetate, ethyl lactate, methyl 3-methoxypropionate, and ethyl 3-ethoxypropionate; propylene glycol diacetate; Polyhydric alcohol carboxylate solvents; polyhydric alcohol partial ether carboxylate solvents such as propylene glycol monomethyl ether acetate and propylene glycol monoethyl ether acetate;
Ketone solvents, for example, acetone, methyl ethyl ketone, diethyl ketone, methyl isobutyl ketone, methyl amyl ketone, diisobutyl ketone, cyclopentanone, cyclohexanone, cycloheptanone;
Is 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 Standard (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 a range of preferably 30 to 90% by mass, more preferably 40 to 88% by mass, and 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 in the pattern film and the thickness of the pattern film are each independently usually 0.1 μm to A 1 mm pattern film can be formed. Such a pattern film is preferable from the viewpoint of cell culture, for example, it can form a cell mass in which cells are three-dimensionally aggregated, and has a small development residue and can contribute to the orientation culture of cells.

  In addition, when the pattern film of the present invention is used, cultured cells that are two-dimensionally arranged, for example, formed in rows, can be favorably obtained. The advantages expected from the two-dimensional arrangement include, for example, a point that a biomimetic (such as a liver shape) can be expected to be performed more accurately.

[Method of manufacturing pattern film]
The method for producing a pattern film according to the present invention comprises the steps of: (1) forming a resin film of the radiation-sensitive resin composition of the present invention on a substrate; (2) exposing the resin film; Step (3) of developing the film with a developer.

  In the present invention, the steps (1) to (3) can be performed at a temperature of preferably 200 ° C. or lower, more preferably 150 ° C. or lower, and further 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.

<Step (1)>
In the step (1), for example, the resin composition of the present invention is applied on a substrate and dried to form a resin film. The drying is performed using an oven or a hot plate, for example, usually at 80 to 150 ° C., preferably 80 to 120 ° C., usually for 1 to 5 minutes. The thickness of the resin film is usually 0.1 μm to 1 mm, preferably 0.1 to 50 μm, more preferably 0.1 to 10 μm. When the film thickness is insufficient, the 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 composed 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, etc .; silicon wafer substrate. A resin substrate made of a biodegradable polymer such as polylactic acid, polyglycolic acid, and polycaprolactane may be used.

  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, a honeycomb structure film described in JP-A-2002-335949 or a three-dimensional culture plate manufactured by ORGANOGENIX Co., Ltd. may be used. An example is a lower substrate from which a frame is removed.

  The substrate surface 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 is mentioned, and the mesh structure is, for example, a polygonal shape 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-adhesive surface from the viewpoint of holding cells and forming a cell tissue body. 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. This is the surface on which the polymer is fixed.

  The functional group having an electric charge can be introduced by treating the substrate surface with plasma or the like. As described above, the substrate may be a plasma processing substrate. Examples of the polymer having cell adhesiveness include, for example, polyacrylic acid, polyvinyl sulfate, polystyrene sulfonic acid, synthetic polymers having a charge such as polyallylamine, chondroitin sulfate, dermatan sulfate, dextran sulfate, keratan sulfate, heparan sulfate, Examples include a charged polysaccharide such as hyaluronic acid and chitin, a cell adhesive protein such as collagen, gelatin, fibronectin, hydronectin, and laminin, and a synthetic polymer in which a cell adhesive protein or a cell adhesive peptide is immobilized.

<Step (2)>
In the step (2), at least a part of the resin film is selectively exposed through 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 light, far ultraviolet light, an electron beam, and X-ray, and light having a wavelength of 200 to 500 nm (eg, i-ray (365 nm)) is preferable. The exposure dose 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 dose is usually 100 to 10,000 mJ / cm ² .

  In order to further promote the crosslinking reaction, it is preferable to perform a heat treatment after the 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, for example, usually 80 to 200 ° C, preferably 80 to 150 ° C, and more preferably 100 to 150 ° C. At about 120 ° C., it is usually about 0.5 to 10 minutes, preferably about 1 to 5 minutes. Within the above-mentioned temperature range, it is possible to suppress adverse effects such as thermal decomposition on the betaine structure of the polymer (A), which is preferable, and also preferable from the viewpoint of patterning performance.

<Step (3)>
In the step (3), a pattern film having a desired pattern is formed on the substrate by developing the exposed resin film with a developer and dissolving / removing a non-exposed portion in the case of a negative type. The size of the non-existing portion of the film in the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 750 μm, more preferably 0.1 to 500 μm. The lower limit of the size of the non-existing 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 developer, 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, for example, alcohols such as methanol, ethanol, n-butanol, propylene glycol monomethyl ether, and ethyl lactate, and ketones such as cyclopentanone. And the like. 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 preferred.

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

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

The polymer may be the polymer (A) described above, 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-existing portion of the film in the pattern film is usually 0.1 μm to 1 mm, preferably 0.1 to 750 μm, more preferably 0.1 to 500 μm. The lower limit of the size of the non-existing 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 film having a configuration not less than the lower limit is preferable from the viewpoint of cell culture, for example, a cell mass in which cells are three-dimensionally aggregated.

The size of the non-existing portion of the film is, for example, a size when a portion where the substrate is exposed is viewed in a plan view without a film containing a polymer having the structural unit (I), That is, the dimension of the exposed substrate surface. For example, when the non-existing portion of the film is a concave portion having a circular shape, an elliptical shape, or a polygonal shape, it is the maximum length on the substrate surface exposed in the concave portion, and the non-existing portion of the film is a linear shape. In the case of a concave portion, it is the distance between the films (the length of the exposed substrate surface in the direction perpendicular to the line). The same applies to the size mentioned elsewhere in this specification.
The pattern substrate of the present invention can be manufactured by the above-described 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 prevention 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 of a surface modifier or a cell adhesion inhibitor.
The surface to which the surface modifier or cell adhesion inhibitor of the present invention is applied is, for example, a surface of various instruments and devices, and specific examples of the instruments and devices include [instruments or devices whose surfaces are modified. Apparatus].

  From the viewpoint of cytotoxicity, 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, based on the cytotoxicity. % By mass or more is more preferable.

  The surface modifier or cell adhesion inhibitor of the present invention may contain, in addition to the polymer (A), at least one selected from solvents, disinfectants and preservatives. Examples of the solvent include water and the solvents described in the column of <Method for Preparing Composition> in addition to water. 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.

  Preventing cell adhesion refers to preventing or suppressing adhesion between a cell and various surfaces with which the cell contacts. The reason why this effect is achieved is not necessarily clear, but the structural units (I) of the polymer (A) hydrophilize the walls of instruments and devices to which the agent is applied, and prevent or suppress cell adhesion. It is presumed that it can be done. In addition to cells, it can also contribute to suppressing the adhesion of biological materials and biological 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 and biotechnology fields (clinical testing and diagnostic agents), for example, clinical diagnostic agents, clinical diagnostic devices, biochips, cell culture instruments, It is useful as a coating agent for materials (solids, instruments, devices, etc.) that come into contact with biological materials, biological materials, etc .; a conditioning agent for measuring cells for fully automatic analyzers used for blood tests and other diagnostics; .

  By coating the surface modifying agent or cell adhesion inhibitor of the present invention on at least a part of an instrument or an apparatus, cells are hardly killed when used, and cells are hardly adhered, and the surface is modified. Tools and devices can be provided.

[Surface modified equipment or device]
The surface-modified appliance or device of the present invention has the above-mentioned polymer (A) on at least a part of its surface (which may be any of an inner wall surface and an outer wall surface). It has a cell adhesion preventing film containing (A).

As instruments and devices, those for cell culture and medical use are preferred.
Examples of the device include a cell culture device; a device for collecting or sending a biological material or a biological tissue (collectively referred to as “biological material or the like”), such as an injection needle, a catheter, and a blood glucose meter; Containers for storing biological materials, such as bags and test tubes; biological materials, such as microchannel devices, microscope peripherals, microwell plates, assay chips, biochips, and measurement cells for fully automatic analyzers. Instruments for analysis; Biotreatment instruments such as reaction vessels, transfer tubes, transfer pipes, purification instruments and cell culture plates; In vivo, such as implants, bone anchors, sutures, anti-adhesion membranes and artificial blood vessels Devices for implantation; other drug delivery vehicles such as vesicles, microparticles and nanoparticles, gastric cameras, microfibers and nanofibers And magnetic particles, and the like.

  Examples of the device include medical devices such as clinical diagnostic devices, biosensors, cardiac pacemakers and implantable biochips; fermentation units and bioreactors.

  The surface-modified device or device of the present invention can be produced 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 manufactured by coating the surface modifier or cell adhesion inhibitor of the present invention on at least a part of the surface of the device or device. The part is preferably a site where the instrument or device comes into contact with cells when the instrument or device is used.

  Examples of the coating method include methods such as coating, spraying, and vapor deposition. Further, the agent may contain a polymerization initiator and a polymerizable compound, and after coating the agent on an apparatus or a device, the polymerizable compound may be polymerized by heat or light. Further, the agent can be cured by using a crosslinking agent and a crosslinking monomer.

  The coating conditions include, for example, a method in which a surface modifier or a cell adhesion inhibitor is brought into contact with an instrument or an apparatus at 50 to 200 ° C. for about 1 to 10 minutes, followed by washing with water and drying.

[Cell culture equipment]
One embodiment of the cell culture device of the present invention has a pattern film obtained by the above-mentioned manufacturing method on a part of its surface. For example, the cell culture instrument of the present invention has a substrate for cell culture, and a pattern film obtained by the manufacturing method on a part of the surface of the substrate. The substrate for cell culture has been described above.

  One embodiment of the cell culture device of the present invention has a substrate and a conventionally known pattern film, and has a polymer (A) on at least a part of the surface of the pattern film. A cell adhesion preventing film containing (A) is provided on at least a part of a surface (particularly, a side wall of the pattern film) that defines a non-existing portion of the pattern film (hereinafter, also referred to as a “cell culture recess” or simply a “recess”). The surface is hydrophilized, making it difficult for cells to adhere to the surface.

  The cell culture instrument has one or more cell culture recesses for holding cells and cell tissue bodies (particularly cell clumps) formed from the cells. The pattern film is a partition that defines the concave portion, and can control the size of the cell tissue, and thus can produce a favorable cell tissue.

  The bottom surface of the recess is formed by the surface of the substrate, and the side surface of the recess is formed by the pattern film. It is preferable that the surface of the substrate has appropriate cell adhesion from the viewpoint of cell retention. The partition walls preferably have cell adhesion preventing properties in consideration of the ease with which the cell tissue body is removed from the recess after the culture. Since the radiation-sensitive resin composition of the present invention contains the polymer (A) for preventing cell adhesion, it is possible to form a partition having the above-mentioned properties.

  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 concave portion is appropriately determined depending on the size and type of the cells and the cell body.

  The size of the concave portion 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, and more preferably 20 μm. The depth of the concave portion 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 tissue can be favorably held in the area defined by the partition.

[Micro channel device]
One embodiment of the microchannel device of the present invention has the pattern film obtained by the above manufacturing method on a part of the inner surface of the microchannel. For example, the pattern film defines a microchannel.

  One embodiment of the microchannel device of the present invention has the polymer (A) on at least a part of the inner surface of the microchannel, and for example, has a cell adhesion preventing film containing the polymer (A). It is provided on at least a part of the inner surface of the road, and preferably on almost the entire surface. The inner surface of the flow channel is hydrophilized, so that biological substances and the like hardly adhere to the surface.

  Examples of the microchannel device include microreaction devices (eg, microreactors and microplants), integrated nucleic acid analysis devices, microelectrophoresis devices, microchromatography devices, and other microanalysis devices; mass spectra, liquid chromatography, and the like. Microdevices for preparation of 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 , A chromatographic chip, a cell analysis chip, a micro-channel chip such as a pharmaceutical screening chip. Among these, a microchannel chip is preferable.

  The micro flow path is a site through which a minute amount of sample (preferably a liquid sample) flows, and the width and depth of the flow path are not particularly limited, but are usually about 0.1 μm to 1 mm, and preferably about 10 μm to 1 μm. 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-described cell culture device to form a cell mass. When culturing a cell, the cell is cultured in a region defined by a partition in the cell culture instrument to form a cell mass. For example, a culture solution containing cells is placed on a cell culture instrument, and the cells seeded on the bottom surface of the recess form a three-dimensionally bonded cell mass on the surface of the substrate that is the bottom surface. The cell mass is cultured over a long period of time while being stably held in the region partitioned by the partition wall in a state of being adhered to the bottom surface of the concave portion without floating in the culture solution.

As the cells, animal species and types of organs / tissues are not particularly limited as long as they form bonds between the cells. Examples of the cells include anchorage-dependent cells. For example, primary cells and cell lines collected from liver, pancreas, kidney, nerve, skin, bone marrow, and the like derived from animals such as humans, pigs, dogs, rats, and mice, and 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, mesenchymal stem cells (eg, UE7T-13 cells); kidney cells such as HEK293 cells; nerve cells such as NT2 cells; Endothelial cells such as UV♀2 cells and HMEC-1 cells; cardiomyocytes such as H9c2 cells; epithelial cells such as Caco-2 cells; and cells obtained by subjecting them to genetic manipulation or the like. In addition, floating cells such as blood cells such as white blood cells, red blood cells, and platelets may be used. One type of cell may be used alone, or two or more types may be used in combination.
As the culture solution, an aqueous solution containing necessary salts and / or nutrients at an appropriate concentration can be used so that the survival state 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 following description of Examples, “parts” means “parts by mass” unless otherwise specified.
The measuring method of each 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 (Showa Denko)
GPC column: Binding GPC-KF-801, GPC-KF-802, GPC-KF-803 and GPC-KF-804 (manufactured by Shimadzu GLC)
Mobile phase: dimethylformamide Column temperature: 40 ° C
Flow rate: 1.0 mL / min Sample concentration: 1.0% by mass
Sample injection volume: 100 μL
Detector: Differential refractometer Standard substance: Monodisperse polystyrene

[Content ratio of structural unit]
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 the examples and comparative examples are shown below.
[Polymerizable compound giving structural unit (I) or comparative structural unit (cI)]
In the synthesis examples, the polymerizable compounds (1-1) to (1-4) which lead to the structural units represented by the following formulas (I-1) to (I-4), (cI-1) to (cI-3) ), (C1-1) to (c1-3).

Ｍｅはメチル基である。

Me is a methyl group.

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

[Polymerizable compound giving structural unit (III)]
In the synthesis examples, polymerizable compounds (3-1) to (3-5) which lead to 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 condenser 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- Charging 4.0 parts of the polymerizable compound (3-3) to give 3), substituting with nitrogen, raising the temperature of the solution to 70 ° C. with gentle stirring, and maintaining the temperature for 4 hours to carry out polymerization. As a result, a polymer solution containing the polymer (A-1) was obtained. The solid content concentration of this polymer solution was 21% by mass, the Mw of the polymer (A-1) was 4,600, and the molecular weight distribution (Mw / Mn) was 2.3.

[Synthesis Examples 2 to 32] (Synthesis of polymer)
The same operations as in Synthesis Example 1 were carried out except that the types and amounts of the polymerizable compounds shown in Table 1 were 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 Mw and Mw / Mn of each polymer were almost the same as those 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) is added, and the solid content concentration becomes 20% by mass. After adding ethyl lactate as a solvent, the mixture was filtered through a membrane filter having a pore size of 0.2 μm to prepare a resin composition (1).

[Examples 2 to 32 and Comparative Examples 1 to 4]
Each resin composition was prepared in the same manner as in Example 1 except that the polymer shown in Table 1 was used as the polymer (A). However, as a 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. Table 1 shows the evaluation results.
[Storage stability performance]
The viscosity at 25 ° C. of the resin composition prepared above was measured using an ELD viscometer manufactured by Tokyo Keiki Co., Ltd. Thereafter, while the composition was allowed to stand at 5 ° C, 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. The number of days is "1" when the number of days is 7 days or less, "2" when the number of days is 8 to 14 days, "3" when the number of days is 15 to 21 days, and 22 to 28 days. The case was evaluated as “4”, and the case of 29 days or more was evaluated as “5”. Table 1 shows the evaluation results. It can be said that the larger the numerical value, the less the thickening and the better the storage stability.

[Patterning performance]
Each of the resin compositions was applied on a glass substrate using a spin coater, and then prebaked on a hot plate at 90 ° C. for 2 minutes to form a coating film having a thickness of 3.0 μm. The obtained coating film was irradiated with ultraviolet rays from a mercury lamp through a pattern mask having a dot pattern having a diameter of 20, 30, 40, 50, 60, 70, 80, 90, or 100 μm. The exposure amount at a wavelength of 365 nm was 300 mJ / cm ² . Next, after baking on a hot plate at 110 ° C. in Examples 1 to 31 and Comparative Examples 1 to 4 and 180 ° C. in Example 32 for 2 minutes, development processing was performed at 25 ° C. for 120 seconds using ultrapure water. . At this time, “1” indicates that no pattern contrast was obtained, “2” indicates that the pattern contrast was obtained but the “missing” portion of the pattern was not completely removed to the bottom of the coating film, and “1” was applied. "3" when the minimum pattern size that escapes to the film bottom is 60 to 100 m, "4" when the minimum pattern size where the pattern "hear" part escapes to the bottom of the coating film is "4", and the pattern "hear" area is painted. The case where the minimum pattern size that passes through to the film bottom is 20 μm was evaluated as “5”. Table 1 shows the evaluation results. It can be said that the larger the numerical value of the evaluation, the smaller the size of the pattern can be formed, and the better the patterning performance.

[Cell aggregation]
For culture of UE7T-13 cells (JCRB1154), which are human bone marrow-derived mesenchymal stem cells (hMSCs), 10% by mass FBS (GE Healthcare), 100 U / mL penicillin, and 100 μg / mL streptomycin (Thermo Fisher Scientific) Dubecco's modified Eagle's medium (Wako Pure Chemical Industries, Ltd.) containing Fick Co., Ltd.). The cells were maintained by performing adhesion culture using a cell culture flask (Thermo Fisher Scientific) in an environment of 37 ° C., a volume fraction of 5% CO ₂ , and 95% air. When the cells were used in the experiment, cells in the logarithmic growth phase were detached by TrypLE ^™ Express (Thermo Fisher Scientific), and the cells were suspended in a medium to prepare a single cell suspension. A glass substrate having a thin film having a thickness of 2 μm in which 100 hole patterns having a diameter of 100 μm were formed in the same manner as in [Patterning performance] was placed on a cell culture container obtained by fractionating in a silicon chamber (flexiPERM micro) manufactured by Sarstad. 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 5% CO ₂ and 95% air.

  Four days later, the collection of cells in the hole pattern was observed in a bright field using an optical microscope (Olympus, IX71). The cells are indicated as “○” when cells are present only in the hole, “△” when most cells are present in the hole, and “×” when cells are present in the hole and other parts without distinction. Assemblability was evaluated. "-" Indicates that a hole pattern could not be formed, and thus cannot be evaluated. Table 1 shows the evaluation results.

  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, while having a quaternary ammonium salt. The radiation-sensitive resin composition comprising a polymer had poor cell aggregation performance characteristics.

Claims (14)

A polymer (A) having a structural unit (I) represented by the formula (1),
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, R ² and R ³ are each independently an alkyl group or an aryl group having 1 to 5 carbon atoms, R ⁴ And R ⁵ are each independently an alkanediyl group having 1 to 10 carbon atoms, X is a carbanion (—COO ⁻ group), a sulfonion (—SO ₃ ⁻ group), or a phosphate anion; Is an oxygen atom, an ester bond, an amide bond, an arylene group, an alkanediyl group having 1 to 10 carbon atoms, or a combination of these groups. ]   The radiation-sensitive resin composition according to claim 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.   The radiation-sensitive resin composition according to claim 2, wherein the structural unit (II) includes a cyclic ether structure, and the cyclic ether structure is at least one selected from an oxirane structure and an oxetane structure.   The radiation-sensitive resin composition according to any one of claims 1 to 3, wherein a content ratio of the structural unit (I) is 10 to 70 mol% based on 100 mol% of all the structural units of the polymer (A). .   The radiation-sensitive resin composition according to any one of claims 1 to 4, wherein the radiation-sensitive compound (B) is contained in an amount of 10 parts by mass or less based on 100 parts by mass of the polymer (A).   The radiation-sensitive resin composition according to any one of claims 1 to 5, 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 6 on a substrate;
(2) A method for producing a pattern film, comprising: exposing the resin film to light; and (3) developing the exposed resin film with a developer.   The method according to claim 7, wherein the developer used in the step (3) contains water.   9. The method according to claim 7, wherein the steps (1) to (3) are performed at a temperature of 200 ° C. or less. 10.   A pattern film formed by the method of manufacturing a pattern film according to claim 7.   A cell culture instrument having the pattern film according to claim 10 on a part of its surface.   A microchannel device having the pattern film according to claim 10 on a part of the inner surface of the microchannel. A substrate, a pattern substrate having a pattern film on the substrate,
A polymer in which the size of the non-existent portion of the film in the pattern film and the thickness of the pattern film are each independently 0.1 μm to 1 mm, and the pattern film has the structural unit (I) represented by the formula (1) A patterned substrate containing:

[In formula (1), R ¹ is a hydrogen atom, a methyl group or a trifluoromethyl group, R ² and R ³ are each independently an alkyl group or an aryl group having 1 to 5 carbon atoms, R ⁴ And R ⁵ are each independently an alkanediyl group having 1 to 10 carbon atoms, X is a carbanion (—COO ⁻ group), a sulfonion (—SO ₃ ⁻ group), or a phosphate anion; Is an oxygen atom, an ester bond, an amide bond, an arylene group, an alkanediyl group having 1 to 10 carbon atoms, or a combination of these groups. ]   A method for producing a cell mass, comprising a step of culturing cells using the cell culture device according to claim 11 to form a cell mass.