JP2002347107A - Stretched film and cell culture base material using the same - Google Patents
Stretched film and cell culture base material using the sameInfo
- Publication number
- JP2002347107A JP2002347107A JP2001152364A JP2001152364A JP2002347107A JP 2002347107 A JP2002347107 A JP 2002347107A JP 2001152364 A JP2001152364 A JP 2001152364A JP 2001152364 A JP2001152364 A JP 2001152364A JP 2002347107 A JP2002347107 A JP 2002347107A
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- Prior art keywords
- stretched film
- polymer
- stretching
- organic solvent
- honeycomb structure
- Prior art date
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Manufacture Of Macromolecular Shaped Articles (AREA)
- Shaping By String And By Release Of Stress In Plastics And The Like (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Description
【発明の詳細な説明】DETAILED DESCRIPTION OF THE INVENTION
【０００１】[0001]
【発明の属する技術分野】本発明は、延伸フィルムおよ
びそれを用いた細胞培養基材に関するものである。より
詳細には、本発明は、医療やバイオテクノロジーで近年
研究が盛んな細胞工学や組織工学に応用可能な延伸フィ
ルムおよびそれを用いた細胞培養基材に関するものであ
り、より具体的には、細胞の培養や、細胞から３次元組
織体の形成のために利用可能な延伸フィルムおよびそれ
を用いた細胞培養基材に関するものである。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a stretched film and a cell culture substrate using the same. More specifically, the present invention relates to a stretched film and a cell culture substrate using the same, which can be applied to cell engineering and tissue engineering, which has been actively studied in recent years in medicine and biotechnology, and more specifically, The present invention relates to a stretched film that can be used for culturing cells and to form a three-dimensional tissue body from cells, and a cell culture substrate using the same.
【０００２】[0002]
【従来の技術】細胞と材料との相互作用において、細胞
は材料表面の化学的な性質のみならず微細な形状によっ
ても影響を受けることが知られている。そこで組織工学
などの観点から細胞の機能制御を目指すとき、細胞と接
触する材料表面の化学的性質と微細な構造の双方の加工
が重要となる。表面の微細加工法としては、表面加工技
術として半導体産業等に利用されているマイクロパター
ン技術を利用した細胞接着面のサイズコントロール、培
養基板への微小溝構造の導入、マイクロスフィアによる
微細凹凸の作製などが挙げられ、表面微細構造が細胞の
成長等に大きく影響を及ぼすことが知られている。2. Description of the Related Art In the interaction between cells and materials, it is known that cells are affected not only by the chemical properties of the material surface but also by the minute shape. Therefore, when aiming at controlling the function of cells from the viewpoint of tissue engineering, it is important to process both the chemical properties and the fine structure of the material surface in contact with the cells. The surface microfabrication method uses the micropattern technology used in the semiconductor industry as a surface processing technology to control the size of the cell adhesion surface, introduce a microgroove structure on the culture substrate, and create microscopic irregularities by microspheres It is known that the surface microstructure greatly affects cell growth and the like.
【０００３】これらのマイクロパターン技術を使った表
面設定は、非常に高度な技術が必要であり、大量生産が
出来ない、高コストになる、などの多くの問題を抱えて
いるのが現状である。全く別の表面パターニング技術と
しては特殊な構造を有するポリマーの希薄溶液を高湿度
下でキャストすることでμｍスケールのハニカム構造を
有するフィルムが得られることが知られている。本方法
はパターニングするに当たっての経済性に優れることが
特徴である。[0003] Surface setting using these micropattern techniques requires very advanced techniques and presents many problems such as the inability to mass-produce and the high cost. . As a completely different surface patterning technique, it is known that a film having a honeycomb structure of a μm scale can be obtained by casting a dilute solution of a polymer having a special structure under high humidity. The feature of this method is that it is economical in patterning.
【０００４】具体的には、サイエンス、1999年、283
巻、ページ373には親水性ブロックと疎水性のブロック
からなるロッド−コイルジブロックポリマーであるポリ
フェニルキノリン−ブロック−ポリスチレンを使う例
が、また、ネイチャー、1994年、369巻、ページ387には
ポリスチレンと剛直なブロックであるポリパラフェニレ
ンとからなるジブロックポリマーを使った例が開示され
ている。このように、従来の技術では自己凝集力の強い
部分と柔軟性を発現する部分とを併せ持つ特殊なポリマ
ーを利用し、これらのポリマーを疎水性有機溶媒に溶解
し、これをキャストする事でハニカム構造体を調製して
いた。一方、本発明者らはシンソリッド フィルムズ、
1998年、327-329巻、ページ854、スープラモレキュラー
サイエンス、1998年、第５巻、ページ331、及びモレキ
ュラー・クリスタル・リキッド・クリスタル、1998年、
第322巻、ページ305に親水性のアクリルアミドポリマー
を主鎖骨格とし、疎水性側鎖としてドデシル基と親水性
側鎖としてラクトース基或いはカルボキシル基を併せ持
つ両親媒性ポリマー、或いはヘパリンやデキストラン硫
酸などのアニオン性多糖と4級の長鎖アルキルアンモニ
ウム塩とのイオンコンプレックスが同様な方法でハニカ
ム構造を有する薄膜を与えることを報告している。[0004] Specifically, Science, 1999, 283
Volume, page 373, uses polyphenylquinoline-block-polystyrene, a rod-coil diblock polymer consisting of a hydrophilic block and a hydrophobic block, and Nature, 1994, Vol. 369, page 387. An example using a diblock polymer composed of polystyrene and polyparaphenylene which is a rigid block is disclosed. As described above, in the conventional technology, a special polymer having both a portion having strong self-cohesive force and a portion exhibiting flexibility is used, and these polymers are dissolved in a hydrophobic organic solvent and cast to form a honeycomb. The structure was being prepared. On the other hand, the present inventors, Shinsolid Films,
1998, Vol. 327-329, page 854, Supra Molecular Science, 1998, Vol. 5, page 331, and Molecular Crystal Liquid Crystal, 1998,
Vol. 322, page 305 shows that an amphiphilic polymer having a hydrophilic acrylamide polymer as a main chain skeleton, a dodecyl group as a hydrophobic side chain and a lactose group or a carboxyl group as a hydrophilic side chain, or heparin or dextran sulfate. It is reported that an ionic complex of an anionic polysaccharide and a quaternary long-chain alkylammonium salt gives a thin film having a honeycomb structure in a similar manner.
【０００５】しかしながらこれらのポリマーでは、得ら
れるハニカム構造体の自己自立性に劣ったり、経時的に
ハニカム構造が崩壊するなどの欠点を有するため、細胞
培養用基材として十分な機能を提供するものでなかっ
た。However, these polymers have drawbacks such as poor self-sustainability of the obtained honeycomb structure and collapse of the honeycomb structure with time, and thus provide a sufficient function as a substrate for cell culture. Was not.
【０００６】細胞工学、組織工学等において細胞培養を
行う時、細胞の足場となる基材が必要であり、前述の如
く、細胞との相互作用において細胞は最良表面の化学的
な性質のみならず微細な形状によっても影響を受けるこ
とが知られている。細胞の機能制御を目指すとき、細胞
と接触する材料表面の化学的性質と細胞の微細な構造の
双方の設計が重要となる。ハニカム構造を有する多孔性
フィルムではハニカムパターンが細胞接着面を提供し、
多孔質構造が細胞の支持基盤へのアクセス、栄養の供給
ルートとなることが示されている。[0006] When cell culture is performed in cell engineering, tissue engineering, etc., a base material serving as a cell scaffold is required. As described above, cells interact not only with the best surface chemical properties but also with the best surface chemistry. It is known that it is affected by a minute shape. When controlling the function of cells, it is important to design both the chemistry of the material surface in contact with the cells and the fine structure of the cells. In a porous film having a honeycomb structure, a honeycomb pattern provides a cell adhesion surface,
It has been shown that the porous structure provides a route for cells to access the support base and supply nutrients.
【０００７】このハニカム構造フィルムをベースに細胞
を組織化すれば、その１つの利用方法として人工臓器が
考えられる。しかし人工臓器等にしたときには体内に埋
め込むことが必須となる為、この基材は長期的には生体
内へ吸収されることが望ましい。これまでのハニカム構
造を与える材料で細胞培養に要する時間は安定に構造を
維持し、それ以上では分解するような生分解性材料から
作られたものはない。言い換えれば、ハニカム構造体と
細胞工学、細胞培養技術を組み合わせ人工臓器等の医療
用途へ展開するに当たっては生分解性材料を使うことが
必須である。[0007] If cells are organized based on the honeycomb structure film, an artificial organ can be considered as one of the utilization methods. However, when it is made into an artificial organ or the like, it is essential to embed it in the body. Therefore, it is desirable that this base material be absorbed into a living body in the long term. There has been no biodegradable material that can maintain a stable structure during the time required for cell culture with a material that provides a honeycomb structure, and degrades beyond that time. In other words, it is essential to use a biodegradable material when developing a medical application such as an artificial organ by combining a honeycomb structure with cell engineering and cell culture technology.
【０００８】このような事情に鑑み、本発明者らは、生
分解性ポリマーが５０〜９９ｗ／ｗ％および両親媒性ポ
リマーが５０〜１ｗ／ｗ％からなるポリマーの疎水性有
機溶媒溶液を、相対湿度５０〜９５％の大気下で基板上
にキャストし、該有機溶媒を徐々に蒸散させると同時に
該キャスト液表面で結露させ、該結露により生じた微小
水滴を蒸発させることで得られるハニカム構造体、並び
に該ハニカム構造体からなるフィルムを提案している
（特願平１１−３４０５６８号明細書（本出願時点にお
いて未公開）。しかしながら、この方法で作製したハニ
カム構造を有するフィルムの細孔の配列は等方的であ
り、この上で細胞の配列を制御することは困難であっ
た。In view of such circumstances, the present inventors have prepared a hydrophobic organic solvent solution of a polymer comprising 50 to 99 w / w% of a biodegradable polymer and 50 to 1 w / w% of an amphiphilic polymer. A honeycomb structure obtained by casting on a substrate in an atmosphere having a relative humidity of 50 to 95%, gradually evaporating the organic solvent and simultaneously dew condensation on the surface of the casting liquid, and evaporating fine water droplets generated by the dew condensation. And a film comprising the honeycomb structure (Japanese Patent Application No. 11-340568 (not disclosed at the time of filing of this application).) However, pores of a film having a honeycomb structure produced by this method are proposed. The arrangement is isotropic, and it was difficult to control the arrangement of cells on this.
【０００９】[0009]
【発明が解決しようとする課題】本発明は、細胞の培養
や、細胞から３次元組織体の形成のために利用可能な延
伸フィルムおよびそれを用いた細胞培養基材を提供する
ことを解決すべき課題とした。さらに本発明は、細胞の
配列を制御することが可能な延伸フィルムおよびそれを
用いた細胞培養基材を提供することを解決すべき課題と
した。SUMMARY OF THE INVENTION An object of the present invention is to provide a stretched film which can be used for culturing cells and for forming a three-dimensional tissue from cells, and a cell culture substrate using the same. Should be an issue. Further, the present invention has an object to provide a stretched film capable of controlling the arrangement of cells and a cell culture substrate using the same.
【００１０】[0010]
【課題を解決するための手段】本発明者らは上記課題を
解決するために鋭意検討を行った結果、生分解性と両親
媒性を併有する弾性ポリマーのハニカム構造体を延伸し
たところ、該延伸により等方的な配列だったハニカム多
孔構造が、異方性を示す細孔の配列に変化することを見
出した。さらに、この延伸フィルム上で心筋細胞の培養
を行った結果、細孔の直線的な配列に沿って細胞を配向
させることができることを実証した。本発明はこれらの
知見に基づいて完成したものである。Means for Solving the Problems The inventors of the present invention have conducted intensive studies in order to solve the above-mentioned problems, and as a result, have drawn a honeycomb structure of an elastic polymer having both biodegradability and amphipathic properties. It has been found that the honeycomb porous structure, which had an isotropic arrangement by stretching, changes to an arrangement of anisotropic pores. Furthermore, as a result of culturing cardiomyocytes on this stretched film, it was demonstrated that the cells could be oriented along a linear array of pores. The present invention has been completed based on these findings.
【００１１】即ち、本発明によれば、生分解性かつ両親
媒性を有する単独のポリマー又は生分解性ポリマーと両
親媒性ポリマーとから成るポリマー混合物の疎水性有機
溶媒溶液を基板上にキャストし、該有機溶媒を蒸散させ
ると同時に該キャスト液表面で結露させ、該結露により
生じた微小水滴を蒸発させることにより得られるハニカ
ム構造体を延伸することによって得られる延伸フィルム
が提供される。That is, according to the present invention, a hydrophobic organic solvent solution of a single biodegradable and amphiphilic polymer or a polymer mixture of a biodegradable polymer and an amphiphilic polymer is cast on a substrate. The present invention provides a stretched film obtained by stretching the honeycomb structure obtained by evaporating the organic solvent and simultaneously forming dew on the surface of the casting liquid and evaporating fine water droplets generated by the dew.
【００１２】本発明において好ましくは、生分解性ポリ
マーとして脂肪族ポリエステルを使用する。本発明にお
いて好ましくは、生分解性ポリマーと両親媒性ポリマー
とから成るポリマー混合物として、５０〜９９ｗ／ｗ％
の生分解性ポリマーおよび５０〜１ｗ／ｗ％の両親媒性
ポリマーからなるポリマー混合物を使用する。In the present invention, an aliphatic polyester is preferably used as the biodegradable polymer. In the present invention, preferably, as a polymer mixture comprising a biodegradable polymer and an amphiphilic polymer, 50 to 99 w / w%
Of a biodegradable polymer and 50-1% w / w amphiphilic polymer.
【００１３】本発明の延伸フィルムは、例えば、疎水性
有機溶媒溶液を基板上にキャストし、高湿度空気を吹き
付けることで該有機溶媒を蒸散させると同時に該キャス
ト液表面で結露させ、該結露により生じた微小水滴を蒸
発させることにより得られるハニカム構造体を延伸する
ことによって得られるか、あるいは、疎水性有機溶媒溶
液を、相対湿度５０〜９５％の大気下で基板上にキャス
トし、該有機溶媒を蒸散させると同時に該キャスト液表
面で結露させ、該結露により生じた微小水滴を蒸発させ
ることにより得られるハニカム構造体を延伸することに
よって得られる。In the stretched film of the present invention, for example, a hydrophobic organic solvent solution is cast on a substrate, and the organic solvent is evaporated by blowing high-humidity air, and at the same time, dew is formed on the surface of the cast liquid. It is obtained by stretching a honeycomb structure obtained by evaporating the generated fine water droplets, or alternatively, a solution of a hydrophobic organic solvent is cast on a substrate in an atmosphere having a relative humidity of 50 to 95%, and the organic organic solvent is dried. Simultaneously with evaporation of the solvent, condensation is formed on the surface of the casting liquid, and the honeycomb structure obtained by evaporating fine water droplets generated by the condensation is obtained by stretching.
【００１４】本発明において好ましくは、延伸は一軸延
伸、二軸延伸又は三軸延伸によって行うことができ、延
伸方向の伸長率は１．１から１０倍の範囲内であり、ま
た、ハニカム構造体の直径は０．１〜１００μｍであ
る。Preferably, in the present invention, the stretching can be performed by uniaxial stretching, biaxial stretching or triaxial stretching, and the elongation in the stretching direction is in the range of 1.1 to 10 times. Has a diameter of 0.1 to 100 μm.
【００１５】本発明の別の側面によれば、上記した本発
明の延伸フィルムからなる細胞培養用基材が提供され
る。本発明のさらに別の側面によれば、上記した本発明
の延伸フィルム又は細胞培養用基材を用いて、細胞を培
養する方法が提供される。According to another aspect of the present invention, there is provided a cell culture substrate comprising the above-described stretched film of the present invention. According to still another aspect of the present invention, there is provided a method of culturing cells using the above-described stretched film or cell culture substrate of the present invention.
【００１６】本発明のさらに別の側面によれば、（１）
生分解性かつ両親媒性を有する単独のポリマー又は複数
のポリマー混合物の疎水性有機溶媒溶液を基板上にキャ
ストし、該有機溶媒を蒸散させると同時に該キャスト液
表面で結露させ、該結露により生じた微小水滴を蒸発さ
せることによりハニカム構造体を延伸する工程；及び
（２）上記のハニカム構造体を延伸する工程：を含む、
本発明の延伸フィルムの製造方法が提供される。According to still another aspect of the present invention, (1)
A hydrophobic organic solvent solution of a single polymer or a plurality of polymer mixtures having biodegradability and amphipathic properties is cast on a substrate, and the organic solvent is evaporated and simultaneously condensed on the surface of the cast liquid. Elongating the honeycomb structure by evaporating the generated fine water droplets; and (2) elongating the above honeycomb structure:
A method for producing a stretched film of the present invention is provided.
【００１７】[0017]
【発明の実施の形態】以下、本発明の実施態様及び実施
方法について詳細に説明する。本発明の延伸フィルム
は、生分解性かつ両親媒性を有する単独のポリマー又は
生分解性ポリマーと両親媒性ポリマーとから成るポリマ
ー混合物の疎水性有機溶媒溶液を基板上にキャストし、
該有機溶媒を蒸散させると同時に該キャスト液表面で結
露させ、該結露により生じた微小水滴を蒸発させること
により得られるハニカム構造体を延伸することによって
得られることを特徴とする。DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, embodiments and a method of implementing the present invention will be described in detail. The stretched film of the present invention is a biodegradable and amphiphilic single polymer or a hydrophobic organic solvent solution of a polymer mixture consisting of a biodegradable polymer and an amphiphilic polymer cast on a substrate,
The organic solvent is evaporated and simultaneously condensed on the surface of the cast liquid, and the honeycomb structure obtained by evaporating fine water droplets generated by the condensation is obtained by stretching.
【００１８】本発明では、生分解性かつ両親媒性を有す
る単独のポリマーを使用してもよいし、あるいは、生分
解性を有するポリマーと両親媒性を有するポリマーから
成る複数のポリマーの混合物を使用してもよい。In the present invention, a single polymer having biodegradability and amphipathic properties may be used, or a mixture of a plurality of polymers consisting of a biodegradable polymer and an amphiphilic polymer may be used. May be used.
【００１９】本発明で用いることができる生分解性ポリ
マーとしては、ポリ乳酸、ポリヒドロキシ酪酸、ポリカ
プロラクトン、ポリエチレンアジペート、ポリブチレン
アジペートなどの生分解性脂肪族ポリエステル、並びに
ポリブチレンカーボネート、ポリエチレンカーボネート
等の脂肪族ポリカーボネート等が、有機溶媒への溶解性
の観点から好ましい。中でも、ポリ乳酸、ポリカプロラ
クトンが入手の容易さ、価格等の観点から望ましい。Examples of the biodegradable polymer that can be used in the present invention include biodegradable aliphatic polyesters such as polylactic acid, polyhydroxybutyric acid, polycaprolactone, polyethylene adipate and polybutylene adipate, and polybutylene carbonate and polyethylene carbonate. Are preferred from the viewpoint of solubility in organic solvents. Among them, polylactic acid and polycaprolactone are desirable from the viewpoint of availability, price, and the like.
【００２０】本発明で用いることができる両親媒性ポリ
マーとしては、細胞培養基材として利用することを考慮
すると毒性のないことが好ましく、ポリエチレングリコ
ール／ポリプロピレングリコールブロック共重合体、ア
クリルアミドポリマーを主鎖骨格とし、疎水性側鎖とし
てドデシル基と親水性側鎖としてラクトース基或いはカ
ルボキシル基を併せ持つ両親媒性ポリマー、或いはヘパ
リンやデキストラン硫酸、核酸（ＤＮＡやＲＮＡ）など
のアニオン性高分子と長鎖アルキルアンモニウム塩との
イオンコンプレックス、ゼラチン、コラーゲン、アルブ
ミン等の水溶性タンパク質を親水性基とした両親媒性ポ
リマー等を利用することが望ましい。The amphiphilic polymer that can be used in the present invention is preferably non-toxic in consideration of its use as a cell culture substrate. A polyethylene glycol / polypropylene glycol block copolymer and an acrylamide polymer are preferably used as the main chain bone. And an amphiphilic polymer having both a dodecyl group as a hydrophobic side chain and a lactose group or a carboxyl group as a hydrophilic side chain, or an anionic polymer such as heparin, dextran sulfate, nucleic acid (DNA or RNA) and a long-chain alkyl. It is desirable to use an ion-complex with an ammonium salt, an amphiphilic polymer having a water-soluble protein such as gelatin, collagen, albumin or the like as a hydrophilic group.
【００２１】また、生分解性かつ両親媒性を有する単独
のポリマーとしては、例えば、ポリ乳酸−ポリエチレン
グリコールブロック共重合体、ポリε−カプロラクトン
−ポリエチレングリコールブロック共重合体、ポリリン
ゴ酸−ポリリンゴ酸アルキルエステルブロック共重合体
などが挙げられる。Examples of the biodegradable and amphiphilic single polymer include polylactic acid-polyethylene glycol block copolymer, polyε-caprolactone-polyethylene glycol block copolymer, polymalic acid-alkyl polymalate. Ester block copolymers and the like can be mentioned.
【００２２】本発明のハニカム構造体を作成するに当た
ってはポリマー溶液上に微小な水滴粒子を形成させるこ
とが必要であることから、使用する有機溶媒としては非
水溶性（疎水性）であることが必要である。疎水性有機
溶媒の例としてはクロロホルム、塩化メチレン等のハロ
ゲン系有機溶媒、ベンゼン、トルエン、キシレン等の芳
香族炭化水素、酢酸エチル、酢酸ブチル等のエステル
類、メチルイソブチルケトンなどの非水溶性ケトン類、
二硫化炭素などが挙げられる。これらの有機溶媒は単独
で使用しても、又、これらの溶媒を組み合わせた混合溶
媒として使用してもよい。疎水性有機溶媒に溶解する生
分解性ポリマーと両親媒性ポリマーの両者の合計のポリ
マー濃度は、好ましくは０．０１から１０重量％であ
り、より好ましくは０．０５から５重量％である。ポリ
マー濃度が０．０１重量％より低いと得られるフィルム
の力学強度が不足し望ましくない。また、ポリマー濃度
が１０重量％以上ではポリマー濃度が高くなりすぎ、十
分なハニカム構造が得られない。In preparing the honeycomb structure of the present invention, since it is necessary to form fine water droplet particles on the polymer solution, the organic solvent used may be water-insoluble (hydrophobic). is necessary. Examples of hydrophobic organic solvents include halogen-based organic solvents such as chloroform and methylene chloride; aromatic hydrocarbons such as benzene, toluene and xylene; esters such as ethyl acetate and butyl acetate; and water-insoluble ketones such as methyl isobutyl ketone. Kind,
Carbon disulfide and the like. These organic solvents may be used alone or as a mixed solvent combining these solvents. The total polymer concentration of both the biodegradable polymer and the amphiphilic polymer dissolved in the hydrophobic organic solvent is preferably 0.01 to 10% by weight, more preferably 0.05 to 5% by weight. When the polymer concentration is lower than 0.01% by weight, the mechanical strength of the obtained film is insufficient, which is undesirable. On the other hand, if the polymer concentration is 10% by weight or more, the polymer concentration becomes too high, and a sufficient honeycomb structure cannot be obtained.
【００２３】また、生分解性ポリマーと両親媒性ポリマ
ーを使用する場合、その組成比は特に限定されないが、
好ましくは９９：１〜５０：５０（wt／wt）の範囲内で
ある。両親媒性ポリマー比が１以下の場合には、均一な
ハニカム構造が得るのが困難となる場合があり、又、両
親媒性ポリマー比が５０以上では得られるハニカム構造
体の安定性、特に力学的な安定性が低下する場合があ
る。When a biodegradable polymer and an amphiphilic polymer are used, their composition ratio is not particularly limited.
Preferably it is in the range of 99: 1 to 50:50 (wt / wt). If the amphiphilic polymer ratio is 1 or less, it may be difficult to obtain a uniform honeycomb structure. If the amphiphilic polymer ratio is 50 or more, the stability of the obtained honeycomb structure, particularly Stability may decrease.
【００２４】本発明においては該ポリマー有機溶媒溶液
を基板上にキャストしハニカム構造体を調製する。基板
としてはガラス、金属、シリコンウェハー等の無機材
料、ポリプロピレン、ポリエチレン、ポリエーテルケト
ン等の耐有機溶剤性に優れた高分子、水、流動パラフィ
ン、液状ポリエーテル等の液体が使用できる。中でも、
基材に水を使用した場合、該ハニカム構造体の特徴であ
る自立性を生かすことで、該構造体を単独で容易に基板
から取り出すことができ、好適である。In the present invention, the polymer organic solvent solution is cast on a substrate to prepare a honeycomb structure. As the substrate, inorganic materials such as glass, metal, and silicon wafer, polymers having excellent organic solvent resistance such as polypropylene, polyethylene, and polyether ketone, and liquids such as water, liquid paraffin, and liquid polyether can be used. Among them,
When water is used as the base material, the structure can be easily taken out from the substrate by itself by taking advantage of the self-sustainability characteristic of the honeycomb structure, which is preferable.
【００２５】本発明で、ハニカム構造が形成される機構
は次のように考えられる。疎水性有機溶媒が蒸発すると
き、潜熱を奪う為に、キャストフィル表面の温度が下が
り、微小な水の液滴がポリマー溶液表面に凝集、付着す
る。ポリマー溶液中の親水性部分の働きによって水と疎
水性有機溶媒の間の表面張力が減少し、このため、水微
粒子が凝集して１つの塊になろうとするに際し、安定化
される。溶媒が蒸発していくに伴い、ヘキサゴナルの形
をした液滴が最密充填した形で並んでいき、最後に、水
が飛び、ポリマーが規則正しくハニカム状に並んだ形と
して残る。In the present invention, the mechanism for forming the honeycomb structure is considered as follows. When the hydrophobic organic solvent evaporates, the temperature of the surface of the cast fill decreases to remove latent heat, and fine water droplets aggregate and adhere to the polymer solution surface. The surface tension between water and the hydrophobic organic solvent is reduced by the action of the hydrophilic portion in the polymer solution, so that the water fine particles are stabilized as they aggregate into one lump. As the solvent evaporates, hexagonal-shaped droplets line up in a close-packed manner, and finally, water splashes out, leaving the polymer in a honeycomb-like form.
【００２６】従って、該フィルムを調製する環境として
は、（１）疎水性有機溶媒溶液を基板上にキャストし、
高湿度空気を吹き付けることで該有機溶媒を蒸散させる
と同時に該キャスト液表面で結露させ、該結露により生
じた微小水滴を蒸発させる方法；並びに（２）疎水性有
機溶媒溶液を、相対湿度50〜95％の大気下で基板上にキ
ャストし、該有機溶媒を蒸散させると同時に該キャスト
液表面で結露させ、該結露により生じた微小水滴を蒸発
させる方法；などが好ましい。このようにしてできるハ
ニカム構造体のひとつひとつ（個々）の大きさは、特に
は限定されないが、好ましくは０．１から１００μmで
あり、より好ましくは０．１から１０μmであり、この
範囲の大きさであれば、細胞培養用の基材として好適に
用いることができる。Therefore, as an environment for preparing the film, (1) a solution of a hydrophobic organic solvent is cast on a substrate,
A method of evaporating the organic solvent by spraying high-humidity air and simultaneously dew condensation on the surface of the casting liquid to evaporate fine water droplets generated by the dew condensation; A method of casting on a substrate in an atmosphere of 95%, evaporating the organic solvent and simultaneously forming dew on the surface of the casting liquid, and evaporating fine water droplets generated by the dew condensation. The size of each (individual) honeycomb structure thus formed is not particularly limited, but is preferably from 0.1 to 100 μm, more preferably from 0.1 to 10 μm, and a size in this range. Then, it can be suitably used as a substrate for cell culture.
【００２７】本発明の延伸フィルムは、上記のようにし
て得られるハニカム構造体を延伸することによって得ら
れる。延伸の方法は、特に限定されず、例えば、ハニカ
ム構造体フィルムの２以上の端をピンセット又は手でつ
まみ、伸長方向に引っ張ることにより行うことができ
る。あるいは、マイクロマニュピレーターを用いて延伸
を行うこともできる。The stretched film of the present invention can be obtained by stretching the honeycomb structure obtained as described above. The stretching method is not particularly limited. For example, the stretching can be performed by pinching two or more ends of the honeycomb structure film with tweezers or a hand, and pulling in the stretching direction. Alternatively, stretching can be performed using a micromanipulator.
【００２８】本発明において、延伸は、一軸延伸、二軸
延伸又は三軸延伸の何れでもよい。本発明における延伸
の具体例の模式図を図１に示す。図１において、（ａ）
は一軸延伸、（ｂ）は二軸延伸、（ｃ）は三軸延伸を示
し、αは対称軸と延伸方向のなす角を示し、β及びγは
延伸方向のなす角を示す。本発明において、延伸方向の
伸長率は特に限定されないが、好ましくは１．１から１
０倍の範囲内である。伸長率が１．１倍以下では延伸に
よる本発明の効果が小さく、また伸長率が１０倍以上で
はフィルムが破壊され易くなる。In the present invention, the stretching may be any of uniaxial stretching, biaxial stretching and triaxial stretching. FIG. 1 is a schematic view of a specific example of stretching in the present invention. In FIG. 1, (a)
Indicates uniaxial stretching, (b) indicates biaxial stretching, (c) indicates triaxial stretching, α indicates the angle between the axis of symmetry and the stretching direction, and β and γ indicate the angles between the stretching directions. In the present invention, the elongation rate in the stretching direction is not particularly limited, but is preferably from 1.1 to 1
It is within the range of 0 times. If the elongation is 1.1 times or less, the effect of the present invention by stretching is small, and if the elongation is 10 times or more, the film is easily broken.
【００２９】上記のようにして作製した本発明の延伸フ
ィルムは、細胞培養用基材として用いることができ、該
基材を用いて細胞を培養することができる。本発明の延
伸フィルムを用いて培養できる細胞の種類は特に限定さ
れず、任意の培養細胞、並びに組織から採取した細胞な
どを培養することができる。The stretched film of the present invention produced as described above can be used as a substrate for cell culture, and cells can be cultured using the substrate. The type of cells that can be cultured using the stretched film of the present invention is not particularly limited, and any cultured cells, cells collected from tissues, and the like can be cultured.
【００３０】細胞培養に用いる培地の種類は特に限定さ
れず、細胞の種類に応じて適当な培地（例えば、Willia
ms’E培地、Ｆ−１０培地、RPM11640培地、EagleのMEM
培地、DMEM培地、またはこれらの培地に牛胎児血清等を
添加した培地等）を選択することができる。培養条件は
細胞の種類に応じて適宜選択することができ、一般的に
はｐＨ６〜８、温度３０〜４０℃、５％ＣＯ2存在下等
の条件下で培養を行うことができる。The type of medium used for cell culture is not particularly limited, and an appropriate medium (for example, Willia
ms'E medium, F-10 medium, RPM11640 medium, Eagle's MEM
Medium, a DMEM medium, or a medium obtained by adding fetal bovine serum or the like to such a medium). Culture conditions can be appropriately selected according to the type of cells, and generally, culture can be performed under conditions of pH 6 to 8, temperature of 30 to 40 ° C., and presence of 5% CO 2 .
【００３１】本発明の細胞培養用基材を用いて、例え
ば、細胞から３次元組織体を形成することもできる。ま
た、本発明の細胞培養用基材は、特には細胞の配列を制
御することが可能であり、特に、心筋組織や血管組織な
どのように細胞の配列構造を有する組織の再生に利用可
能である。以下、本発明を実施例を使って詳細に説明す
るが、本発明は実施例によって何ら限定されるものでは
ない。Using the cell culture substrate of the present invention, for example, a three-dimensional tissue can be formed from cells. Further, the cell culture substrate of the present invention can control the arrangement of cells, in particular, and can be used particularly for regeneration of a tissue having a cell arrangement structure such as a myocardial tissue or a vascular tissue. is there. Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited to the examples.
【００３２】[0032]
【実施例】実施例１：ポリε−カプロラクトン（分子
量：100,000〜190,000）のクロロホルム溶液（１０ｇ／
Ｌ）、両親媒性高分子Ｃａｐ（化学構造は以下に記載）
のベンゼン溶液（０．４ｇ／Ｌ）、ベンゼンを1：２．
５：６．５の割合（体積比）で混合し、ポリマー溶液を
調製した。直径９ｃｍのガラスシャーレにMilli−Q水を
張り、この水面上にポリマー溶液９０μＬを一様に展開
し液滴が拡がらない状態にした。溶媒が蒸発したのを確
認した後、同じ溶液１５０μLを水面上にキャストし、
高湿度空気を吹き付けることでハニカムフィルム（直
径：２ｃｍ）を得た（図２）。水面上に浮いているハニ
カムフィルムの両端をピンセットでつまみ、一軸方向に
延伸し、長軸：３．４ｃｍ、短軸：１．８ｃｍの延伸フ
ィルムを得た。これをガラス板上に移した。延伸フィル
ムの光学顕微鏡写真（図２）に、伸長した細孔の配列構
造が認められた。EXAMPLES Example 1 A chloroform solution of polyε-caprolactone (molecular weight: 100,000 to 190,000) (10 g /
L), amphiphilic polymer Cap (chemical structure is described below)
Benzene solution (0.4 g / L), benzene in 1: 2.
The mixture was mixed at a ratio of 5: 6.5 (volume ratio) to prepare a polymer solution. Milli-Q water was spread on a glass Petri dish having a diameter of 9 cm, and 90 μL of the polymer solution was uniformly spread on the water surface so that droplets did not spread. After confirming that the solvent had evaporated, 150 μL of the same solution was cast on the water surface,
A honeycomb film (diameter: 2 cm) was obtained by blowing high humidity air (FIG. 2). Both ends of the honeycomb film floating on the water surface were pinched with tweezers and stretched uniaxially to obtain a stretched film having a major axis of 3.4 cm and a minor axis of 1.8 cm. This was transferred onto a glass plate. An optical micrograph (FIG. 2) of the stretched film showed an elongated pore array structure.
【００３３】[0033]
【化１】 Embedded image
【００３４】試験例１：実施例１で得られた延伸フィル
ムをガラス基板上に設置し、この上でラット胎児心臓由
来心筋細胞の培養を行った。培養はＦ１０培地を用い、
ＣＯ2インキュベーター内（ＣＯ2濃度＝５％、温度＝３
７℃、相対湿度＝８０％）で行った。比較例として未延
伸のハニカム構造体フィルムをガラス基板上に設置した
もの、細孔のないＰＣＬ−両親媒性高分子キャスト膜を
ガラス基板上に設置したものに心筋細胞を播種し、同様
の条件で培養を行った。 Test Example 1: The stretched film obtained in Example 1 was placed on a glass substrate, and cardiomyocytes derived from rat fetal heart were cultured thereon. Culture uses F10 medium,
In a CO 2 incubator (CO 2 concentration = 5%, temperature = 3
7 ° C., relative humidity = 80%). As comparative examples, cardiomyocytes were seeded on an unstretched honeycomb structure film provided on a glass substrate, and a PCL-amphiphilic polymer cast film having no pores provided on a glass substrate, under the same conditions. Was performed.
【００３５】培養後、光学顕微鏡写真をとった結果を図
２に示す。図２の結果から分かるように、実施例１で得
られた延伸フィルムを用いた場合、細胞は、特定の一方
向に伸展かつ配列しており、その方向はフィルムの延伸
方向に一致していた。また、比較例の場合には、心筋細
胞の伸展および配列方向に規則性は認められなかった。After the culture, the results of taking an optical micrograph are shown in FIG. As can be seen from the results in FIG. 2, when the stretched film obtained in Example 1 was used, the cells were stretched and arranged in one specific direction, and the direction matched the stretching direction of the film. . In the case of the comparative example, no regularity was observed in the extension and arrangement direction of the cardiomyocytes.
【００３６】[0036]
【発明の効果】本発明により、細胞の培養や、細胞から
３次元組織体の形成のために利用可能な延伸フィルムお
よびそれを用いた細胞培養基材、特には細胞の配列を制
御することが可能な延伸フィルムおよびそれを用いた細
胞培養基材を提供することが可能になった。本発明の延
伸フィルムおよびそれを用いた細胞培養基材は、特に、
心筋組織や血管組織などのように細胞の配列構造を有す
る組織の再生に利用可能であり、広範な細胞及び／又は
組織の培養・再生に応用可能である。According to the present invention, it is possible to control a stretched film which can be used for culturing cells and to form a three-dimensional tissue body from cells, and a cell culture base material using the stretched film, in particular, the arrangement of cells. It has become possible to provide a possible stretched film and a cell culture substrate using the same. The stretched film of the present invention and the cell culture substrate using the same are, in particular,
The present invention can be used for regeneration of a tissue having a cell array structure such as a myocardial tissue or a vascular tissue, and can be applied to culture and regeneration of a wide range of cells and / or tissues.
【図１】図１は、ハニカム構造体の延伸の様式を示す図
である。FIG. 1 is a diagram showing a mode of stretching a honeycomb structure.
【図２】図２は、ポリε−カプロラクトンハニカム構造
体（左上）およびその延伸構造体（左下）の光学顕微鏡
写真、並びに、これらの構造体の上で培養した心筋細胞
の蛍光顕微鏡写真（右上：ポリε−カプロラクトンハニ
カム構造体、及び、右下：ポリε−カプロラクトンハニ
カム構造体の延伸フィルム）を示す。FIG. 2 shows optical micrographs of a poly ε-caprolactone honeycomb structure (upper left) and its expanded structure (lower left), and a fluorescence micrograph of cardiomyocytes cultured on these structures (upper right). : Poly-ε-caprolactone honeycomb structure and lower right: stretched film of poly-ε-caprolactone honeycomb structure).
フロントページの続き (72)発明者 新井 景子 埼玉県和光市広沢２番１号 理化学研究所 内 Ｆターム(参考） 4B029 AA08 BB11 CC02 CC08 GA03 GB09 4B065 AA90X BC41 CA44 4F210 AA24 AG01 QA02 QC01 QC05 QC09 QD19 QG01 QG08 QG18Continued on front page (72) Inventor Keiko Arai 2-1 Hirosawa, Wako-shi, Saitama F-term in RIKEN (reference) 4B029 AA08 BB11 CC02 CC08 GA03 GB09 4B065 AA90X BC41 CA44 4F210 AA24 AG01 QA02 QC01 QC05 QC09 QD19 QG01 Q QG18
Claims (11)
リマー又は生分解性ポリマーと両親媒性ポリマーとから
成るポリマー混合物の疎水性有機溶媒溶液を基板上にキ
ャストし、該有機溶媒を蒸散させると同時に該キャスト
液表面で結露させ、該結露により生じた微小水滴を蒸発
させることにより得られるハニカム構造体を延伸するこ
とによって得られる延伸フィルム。1. A hydrophobic organic solvent solution of a biodegradable and amphiphilic single polymer or a polymer mixture comprising a biodegradable polymer and an amphiphilic polymer is cast on a substrate, and the organic solvent is evaporated. A stretched film obtained by stretching a honeycomb structure obtained by causing condensation to form on the surface of the casting liquid and evaporating fine water droplets generated by the condensation.
テルを使用する、請求項１に記載の延伸フィルム。2. The stretched film according to claim 1, wherein an aliphatic polyester is used as the biodegradable polymer.
から成るポリマー混合物として、５０〜９９ｗ／ｗ％の
生分解性ポリマーおよび５０〜１ｗ／ｗ％の両親媒性ポ
リマーからなるポリマー混合物を使用する、請求項１又
は２に記載の延伸フィルム。3. A polymer mixture comprising 50 to 99% w / w biodegradable polymer and 50% to 1% w / w amphiphilic polymer as a polymer mixture comprising a biodegradable polymer and an amphiphilic polymer. The stretched film according to claim 1 or 2, wherein
し、高湿度空気を吹き付けることで該有機溶媒を蒸散さ
せると同時に該キャスト液表面で結露させ、該結露によ
り生じた微小水滴を蒸発させることにより得られるハニ
カム構造体を延伸することによって得られる、請求項１
から３の何れかに記載の延伸フィルム。4. A solution of a hydrophobic organic solvent is cast on a substrate, and the high-humidity air is blown to evaporate the organic solvent and simultaneously cause dew condensation on the surface of the cast liquid, thereby evaporating fine water droplets generated by the dew condensation. The honeycomb structure obtained by stretching is obtained by stretching.
4. The stretched film according to any one of items 1 to 3.
９５％の大気下で基板上にキャストし、該有機溶媒を蒸
散させると同時に該キャスト液表面で結露させ、該結露
により生じた微小水滴を蒸発させることにより得られる
ハニカム構造体を延伸することによって得られる、請求
項１から４の何れかに記載の延伸フィルム。5. The method according to claim 1, wherein the hydrophobic organic solvent solution is adjusted to a relative humidity of 50 to 50.
By casting on a substrate in an atmosphere of 95%, evaporating the organic solvent and simultaneously forming dew on the surface of the casting liquid, and elongating a honeycomb structure obtained by evaporating fine water droplets generated by the dew condensation. The stretched film according to any one of claims 1 to 4, which is obtained.
によって行う、請求項１から５の何れかに記載の延伸フ
ィルム。6. The stretched film according to claim 1, wherein the stretching is performed by uniaxial stretching, biaxial stretching, or triaxial stretching.
範囲内である、請求項１から６の何れかに記載の延伸フ
ィルム。7. The stretched film according to claim 1, wherein the stretch ratio in the stretching direction is in the range of 1.1 to 10 times.
００μｍである、請求項１から７の何れかに記載の延伸
フィルム。8. The honeycomb structure having a diameter of 0.1 to 1
The stretched film according to any one of claims 1 to 7, which has a thickness of 00 µm.
ィルムからなる細胞培養用基材。9. A cell culture substrate comprising the stretched film according to claim 1.
フィルム又は請求項９に記載の細胞培養用基材を用い
て、細胞を培養する方法。10. A method for culturing cells using the stretched film according to claim 1 or the substrate for cell culture according to claim 9.
単独のポリマー又は複数のポリマー混合物の疎水性有機
溶媒溶液を基板上にキャストし、該有機溶媒を蒸散させ
ると同時に該キャスト液表面で結露させ、該結露により
生じた微小水滴を蒸発させることによりハニカム構造体
を延伸する工程；及び（２）上記のハニカム構造体を延
伸する工程：を含む、請求項１に記載の延伸フィルムの
製造方法。11. A solution of a biodegradable and amphiphilic single polymer or a mixture of a plurality of polymers in a hydrophobic organic solvent is cast on a substrate, and the organic solvent is evaporated and at the same time the surface of the cast liquid is evaporated. And stretching the honeycomb structure by evaporating fine water droplets generated by the condensation; and (2) stretching the honeycomb structure. 2. The stretched film according to claim 1, further comprising: Production method.
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| JP2001152364A JP4731044B2 (en) | 2001-05-22 | 2001-05-22 | Stretched film and cell culture substrate using the same |
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| JP2001152364A JP4731044B2 (en) | 2001-05-22 | 2001-05-22 | Stretched film and cell culture substrate using the same |
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| Publication Number | Publication Date |
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| JP4731044B2 JP4731044B2 (en) | 2011-07-20 |
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ID=18997053
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2001152364A Expired - Fee Related JP4731044B2 (en) | 2001-05-22 | 2001-05-22 | Stretched film and cell culture substrate using the same |
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| JP (1) | JP4731044B2 (en) |
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Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4868668A (en) * | 1971-12-17 | 1973-09-19 | ||
| JPS5035271A (en) * | 1973-05-07 | 1975-04-03 | ||
| JPS62234510A (en) * | 1986-04-05 | 1987-10-14 | Asahi Chem Ind Co Ltd | Drying method for regenerated hollow cellulose yarn |
| JPH05339387A (en) * | 1992-06-12 | 1993-12-21 | Res Dev Corp Of Japan | Production of thin crosslinked polymer film |
| JPH06240037A (en) * | 1992-02-06 | 1994-08-30 | Mitsui Toatsu Chem Inc | Thermally shrinkable composite foam sheet |
| JPH08300481A (en) * | 1995-05-08 | 1996-11-19 | Mitsubishi Rayon Co Ltd | Heat-shrinkable film and manufacture thereof |
| JPH09132638A (en) * | 1995-09-07 | 1997-05-20 | Mitsui Toatsu Chem Inc | Polymer capable of being absorbed by living tissue and its production |
| JPH09235456A (en) * | 1996-03-01 | 1997-09-09 | Shin Etsu Chem Co Ltd | Biodegradable film |
| JPH10219008A (en) * | 1997-02-04 | 1998-08-18 | Kagaku Gijutsu Shinko Jigyodan | Thin membrane of nucleic acid polymer pattern, its formation and thin membrane |
| JPH11169199A (en) * | 1997-12-10 | 1999-06-29 | Japan Science & Technology Corp | Gene sensor |
| JP2001049098A (en) * | 1999-08-10 | 2001-02-20 | Mitsui Chemicals Inc | Lactic acid resin composition and molding form |
| JP2001157574A (en) * | 1999-11-30 | 2001-06-12 | Terumo Corp | Honeycomb structure, method for preparing the structure, film and cell culture bade using the structure |
| JP2002514253A (en) * | 1997-03-31 | 2002-05-14 | ザ、リージエンツ、オブ、ザ、ユニバーシティ、オブ、ミシガン | Open pore biodegradable matrix |
| JP2002335949A (en) * | 2001-05-22 | 2002-11-26 | Inst Of Physical & Chemical Res | Cell three-dimensional tissue culture method using honeycomb structure film |
-
2001
- 2001-05-22 JP JP2001152364A patent/JP4731044B2/en not_active Expired - Fee Related
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4868668A (en) * | 1971-12-17 | 1973-09-19 | ||
| JPS5035271A (en) * | 1973-05-07 | 1975-04-03 | ||
| JPS62234510A (en) * | 1986-04-05 | 1987-10-14 | Asahi Chem Ind Co Ltd | Drying method for regenerated hollow cellulose yarn |
| JPH06240037A (en) * | 1992-02-06 | 1994-08-30 | Mitsui Toatsu Chem Inc | Thermally shrinkable composite foam sheet |
| JPH05339387A (en) * | 1992-06-12 | 1993-12-21 | Res Dev Corp Of Japan | Production of thin crosslinked polymer film |
| JPH08300481A (en) * | 1995-05-08 | 1996-11-19 | Mitsubishi Rayon Co Ltd | Heat-shrinkable film and manufacture thereof |
| JPH09132638A (en) * | 1995-09-07 | 1997-05-20 | Mitsui Toatsu Chem Inc | Polymer capable of being absorbed by living tissue and its production |
| JPH09235456A (en) * | 1996-03-01 | 1997-09-09 | Shin Etsu Chem Co Ltd | Biodegradable film |
| JPH10219008A (en) * | 1997-02-04 | 1998-08-18 | Kagaku Gijutsu Shinko Jigyodan | Thin membrane of nucleic acid polymer pattern, its formation and thin membrane |
| JP2002514253A (en) * | 1997-03-31 | 2002-05-14 | ザ、リージエンツ、オブ、ザ、ユニバーシティ、オブ、ミシガン | Open pore biodegradable matrix |
| JPH11169199A (en) * | 1997-12-10 | 1999-06-29 | Japan Science & Technology Corp | Gene sensor |
| JP2001049098A (en) * | 1999-08-10 | 2001-02-20 | Mitsui Chemicals Inc | Lactic acid resin composition and molding form |
| JP2001157574A (en) * | 1999-11-30 | 2001-06-12 | Terumo Corp | Honeycomb structure, method for preparing the structure, film and cell culture bade using the structure |
| JP2002335949A (en) * | 2001-05-22 | 2002-11-26 | Inst Of Physical & Chemical Res | Cell three-dimensional tissue culture method using honeycomb structure film |
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