JP2013504669A - 三次元多孔質構造体 - Google Patents
三次元多孔質構造体 Download PDFInfo
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- JP2013504669A JP2013504669A JP2012529158A JP2012529158A JP2013504669A JP 2013504669 A JP2013504669 A JP 2013504669A JP 2012529158 A JP2012529158 A JP 2012529158A JP 2012529158 A JP2012529158 A JP 2012529158A JP 2013504669 A JP2013504669 A JP 2013504669A
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Abstract
【選択図】 なし
Description
公知の固相担体は通常、用途に合うように特定のサイズ及び物理的性質のポリマー微粒子を含む。使い易さに関しては、これらのポリマー微粒子は球形であることが多く、規定の粒径分布をもつ。微粒子が球形の性質をもつと、ポリマーの流動性及び濾過特性が改善する。固体担体を使用することは好都合であるが、固相アプローチに対しては不都合な点がある。たとえば、ペプチド及びオリゴヌクレオチドの固相合成に通常使用される市販の担体は、複雑な製造プロセスなどのため高価なことがよくある。これらのプロセスは、重合、面倒な粒径分類、たとえば篩分け及び空気分級(air classification)の間に、連続相に対しモノマー損失、様々な粒径の生成、及び面倒な微細粒子の生成などの幾つかの点において不都合な点がある。
三次元で細胞を培養及び分化させるために、組織形成を支持しえる人工構造体に細胞を播種することが必要である。通常、骨格と称されるこれらの構造体は、ex-vivo及びin-vivoの両方でそれ自体の微環境(microenvironment)に影響を与えられるようにするのに重要である。この骨格はまず、細胞付着及び移動(migration)でき、細胞栄養素及び圧搾された廃棄物(expressed waste)を拡散させることができ、必要な機械的及び生物学的特性を提供することができなければならない。流動下での多孔性及び特性は、マクロポーラス構造体を設計する際に考慮する重要な因子である。今まで使用されてきたマクロポーラス骨格は境界のはっきりしない細孔構造であり、細孔は通常小さすぎるか、広い孔径分布を有している。栄養素及び廃棄物の移動が容易で、細胞を迅速に且つ制約なく拡散させられる構造体が望ましい。
液体と微粒子とを接触させる際、使用したモノマーに依存して慣用法にて重合を開始する。フリーラジカルまたは触媒を液体成分に添加して、重合を開始または実施することができる。
構造体内の孔径対孔径出現率(occurrence)のグラフをプロットする際に、公知の多孔性ポリマーは、通常広い“ベル型”ガウシアン分布(Gaussian distribution)を示すであろうが、本発明の構造体は、もっと狭い“スパイク”を示し、狭い孔径分布を示す。
3D構造体の細孔サイズは、粒径により示され、例えば、0.1μm〜1mmなどのサブミクロンサイズからミリメートルスケールでありえる。たとえば、酵母細胞(パン酵母)を使用して、1〜10μmなどの小さな細孔を生成することができ、コーティング業界で使用される球状ワックス微粒子を使用して、5〜50μm範囲のやや大きな細孔を生成することができる。ガラス及びポリマー微粒子は、多様なサイズで入手可能であるので、1〜500μm、特に10〜300μmのサイズをもつ細孔を作るのに所望の粒径を選択することができる。重要な留意事項は、微粒子は、3D構造体の孔径を決定するので狭いサイズ分布をもつということである。
好ましい場合には、3Dマクロポーラスポリマーは、重合の間または重合後のいずれかで担体微粒子に共有結合することができる。あるいは、一種以上の構成要素のモノマーは、重合開始前に微粒子表面に共有結合することができる。
3Dマクロポーラスポリマーは、電子伝導及び発光ポリマーを含む用途で使用することができる。発光ポリマーを含む微粒子体はディスプレイパネル上に配置することができる。
本発明はさらに、クロマトグラフィープロセスでモノリスとしての本発明に従った3Dマクロポーラスポリマーの使用を提供する。
さらなる用途では、本発明の微粒子状担体は、たとえば遷移金属触媒及びリガンドを固定化することにより、化学触媒でも使用することができる。
メチレン-ビス-アクリルアミド(MBA)(123mg,0.8mmol)をN,N-ジメチルホルムアミド(DMF)(1.26cm3)中に温めながら溶解した。この溶液を室温に冷却し、2-(ジメチルアミノ)エチルメタクリレート(DMAEM)(0.43cm3,0.55mmol)、アクリルアミド(2.02g,28.5mmol)、テトラメチレンジアミン(TEMEDA)(0.006cm3,0.05mmol)及び水を添加した。この混合物を全ての成分が溶解するまで攪拌した。Expancel 920 DEX 80 d30(80μmポリアクリルニトリルバルーン)(150mg,10cm3)を添加し、よく攪拌してから、過硫酸アンモニウム溶液(1%w/vの0.285cm3)を添加して重合を開始した。混合物をテフロン(登録商標)型でディスクにキャストした。重合を室温で一晩、放置して完了させて、次いで水でよく洗浄してから凍結乾燥した。
実施例2.ポリアクリル酸の製造
メチレン-ビス-アクリルアミド(MBA)(240mg,1.55mmol)をN,N-ジメチルホルムアミド(DMF)(2.5cm3)に温めながら溶解した。この溶液を室温に冷却し、2-(ジメチルアミノ)エチルメタクリレート(DMAEM)(0.43cm3,0.55mmol)、アクリルアミド(1.9g,26.8mmol)、テトラメチレンジアミン(TEMEDA)(0.006cm3,0.05mmol)及び水を添加した。この混合物を全ての成分が溶解するまで攪拌した。Expancel 920 DEX 80 d30(80μmポリアクリルニトリルバルーン)(150mg,10cm3)を添加し、よく攪拌してから、過硫酸アンモニウム溶液(1%w/vの0.285cm3)を添加して重合を開始した。混合物をテフロン(登録商標)型でディスクにキャストした。重合を室温で一晩、放置して完了させた。
実施例3.ポリスチレン-ジビニルベンゼンの製造
1,4-ジビニルベンゼン(DVB)(4.6cm3,43mmol)、スチレン(0.92cm3,8.85mmol)及び1,1-アゾビス(シクロヘキサン-カルボニトリル)(100mg)をExpancel 920 DEX 80 d30(150mg,10cm3)に添加し、よく攪拌した。この混合物をテフロン(登録商標)型でディスクにキャストした。重合を80℃で一晩、放置して完了させた。ディスクはジエチルエーテルでよく洗浄してから風乾した。
生成物のSEMを図3に示す。
1,4-ジビニルベンゼン(DVB)(0.934cm3,8.7mmol)、スチレン(0.19cm3,1.83mmol)、トルエン(1cm3)及び1,1-アゾビス(シクロヘキサン-カルボニトリル)(50mg)をExpancel 920 DEX 80 d30(150mg,10cm3)に添加し、よく攪拌した。この混合物をテフロン(登録商標)型でディスクにキャストし、重合を80℃で一晩、放置して完了させた。ディスクはジエチルエーテルでよく洗浄してから風乾した。
実施例5.ポリスチレン-ジビニルベンゼンの製造
1,4-ジビニルベンゼン(DVB)(1.63cm3,15mmol)、スチレン(0.92cm3,5.4mmol)、トルエン(1.9cm3)及び1,1-アゾビス(シクロヘキサン-カルボニトリル)(15mg)をQ-Cel 300(〜80μm中空ガラス球)(300mg)に添加した。この混合物をテフロン(登録商標)型でディスクにキャストし、重合を100℃で2.5時間、放置して完了させた。ディスクはジクロロメタンでよく洗浄してから風乾した。
実施例6.シリカの製造
テトラメトキシオルトシラン(TMOS)(2.5cm3)及び3-アミノプロピル-トリメトキシシラン(APTS)(0.025cm3)をエタノール-水(5cm3,95:5v/v)に添加し、よく攪拌した。この混合物をExpancel 920 DEX 80 d30(200mg,13cm3)に添加し、よく攪拌した。この混合物をテフロン(登録商標)型でディスクにキャストし、重合を50℃で数時間、放置して完了させた。ディスクはDMFで約60時間かけて処理し、水でよく洗浄してから凍結乾燥した。
実施例7.中空ガラス球を含むシリカの製造
TMOS(2.5cm3)及びAPTS(0.025cm3)をエタノール-水(2.5cm3,95:5v/v)に添加し、よく攪拌した。この混合物をQ-Cel 7014(約100μm中空ガラス球)に添加し、よく攪拌した。この混合物をテフロン(登録商標)型でディスクにキャストし、重合を50℃で48時間、放置して完了させた。ディスクは水でよく洗浄してから凍結乾燥した。
実施例8.ガラスシードビーズ内に封入された中空ポリアクリルニトリル球を含むポリアクリルアミド
メチレン-ビス-アクリルアミド(MBA)(240mg,1.55mmol)をN,N-ジメチルホルムアミド(DMF)(2.5cm3)中に温めながら溶解した。この溶液を室温に冷却し、2-(ジメチルアミノ)エチルメタクリレート(DMAEM)(0.43cm3,0.55mmol)、アクリルアミド(1.9g,26.8mmol)、テトラメチレンジアミン(TEMEDA)(0.006cm3,0.05mmol)及び水を添加した。この混合物を全ての成分が溶解するまで丸底フラスコ中で攪拌した。Expancel 920 DEX 80 d30(80μmポリアクリルニトリルバルーン)(150mg,10cm3)を添加し、よく攪拌してから、過硫酸アンモニウム溶液(1%w/vの0.285cm3)を添加して重合を開始した。1mm直径のガラスシードビーズ(30cm3)をこの混合物に添加し、フラスコを一時的に真空にして、重合混合物をシードビーズ内に引き込んだ。重合を室温で一晩、放置して完了させた。
生成物のSEMを図8に示す。
実施例2で製造したディスクの断面を使用してヒト胎児幹細胞を培養した。細胞集団をディスクに適用し、慣用の栄養素を与えた。幹細胞は、幹細胞が3D構造体の細孔内に産生するように、増殖し、3D構造体内に広がった。
Claims (17)
- 場合によりポリマーの細孔内に微粒子を有する多孔質ポリマー構造体を含む三次元多孔質ポリマー構造体であって、前記細孔が狭い孔径分布を有する、前記三次元多孔質ポリマー構造体。
- 前記多孔質ポリマーが、ポリアクリルアミド、ポリアクリル酸、ポリスチレン-ジビニルベンゼン、シリカ、ポリスチレン、セルロース、寒天、ポリジメチルアクリルアミド、ポリメチルメタクリレート、ポリメタクリレート、ポリアクリレート、ポリ尿素、ポリアクリロイルモルホリン、ポリビニルアルコール、ポリベータヒドロキシエステル及びポリアクリロニトリル、ポリアルキレングリコール及びポリハイプから選択される、請求項1に記載の構造体。
- 前記構造体の平均孔径の1/10〜10倍の範囲の孔径をもつような構造体中の細孔である、請求項1または2に記載の構造体。
- 前記孔径範囲が、前記平均孔径の75〜125%である、請求項3に記載の構造体。
- 前記構造体の細孔内に微粒子を含む、請求項1〜4のいずれか1項に記載の構造体。
- 前記微粒子は、細胞と、ガラス、セラミック、ポリマー、天然産物及び金属から選択される材料とから選択される、請求項5に記載の構造体。
- 前記粒子が中空である、請求項5または6に記載の構造体。
- 前記微粒子が球状または楕円体である、請求項5〜7のいずれか1項に記載の構造体。
- 前記微粒子が、中空ガラス球、中空ポリマー球、酵母細胞または蝋の球である、請求項5〜8のいずれか1項に記載の構造体。
- 前記微粒子が中空ポリマー含み、多孔質ポリマーに結合するための誘導体と反応するための活性部位をもつ、請求項5〜9のいずれか1項に記載の構造体。
- 触媒、ペプチド合成用の開始剤種、オリゴヌクレオチド合成用開始剤種、固相有機化学用開始剤種、酵素、医薬品、農業化学品、タンパク質若しくは他の生物学的高分子または全細胞から選択されるポリマーで担持された機能性材料をさらに含む、請求項1〜10のいずれか1項に記載の構造体。
- 領域に微粒子を最密充填して、微粒子の三次元配列を提供する工程、重合可能なモノマーまたはモノマーの溶液と前記配列とを、前記モノマーまたは前記溶液が、前記粒子の間の隙間を充填するように接触させる工程、モノマーの重合を実施し、それによって前記微粒子の周りにポリマー構造体を形成する工程、及び場合により、前記微粒子を前記構造体から除去する工程を含む、三次元多孔質構造体の製造プロセス。
- モノマーを含む液体中に微粒子のスラリーを形成する工程、及び前記モノマーを重合させる工程を含む請求項12に記載のプロセス。
- ペプチド、オリゴヌクレオチド、オリゴ糖から選択される種の固相合成;固相抽出;固相有機化学;固相試薬、金属及び他の触媒、生体触媒、酵素、タンパク質、ポリクローナル及びモノクローナル抗体を含む抗体、全細胞並びにポリマーから選択される種の固定化;細胞培養;クロマトグラフィー分離用固定相の調製から選択されるプロセスにおける;または吸着剤として使用するための;または絶縁材料として若しくは組織再生で使用するための、請求項1〜11のいずれか1項に記載の三次元多孔質構造体の使用。
- 細胞または無細胞系から分泌または排出されたモノクローナル抗体または他の生物活性分子を連続生成するためのプロセスにおける、請求項1〜11のいずれか1項に記載の三次元多孔質構造体の使用であって、前記細胞系または無細胞系は三次元構造体上で増殖するか、または三次元構造体に可逆的若しくは非可逆的に結合している、前記使用。
- 有機体用の生産ユニットとしての請求項1〜11のいずれか1項に記載の三次元多孔質構造体の使用。
- 前記有機体が幹細胞、臓器及び骨から選択される、請求項16に記載の使用。
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WO2017104610A1 (ja) * | 2015-12-14 | 2017-06-22 | 国立研究開発法人科学技術振興機構 | 多孔フィルム、多孔フィルム製造方法、マイクロレンズアレイ、マイクロリアクターおよびバイオデバイス |
JPWO2017104610A1 (ja) * | 2015-12-14 | 2018-09-27 | 国立研究開発法人科学技術振興機構 | 多孔フィルム、多孔フィルム製造方法、マイクロレンズアレイ、マイクロリアクターおよびバイオデバイス |
WO2019146558A1 (ja) * | 2018-01-23 | 2019-08-01 | 富士フイルム株式会社 | 多孔成形体 |
WO2019146557A1 (ja) * | 2018-01-23 | 2019-08-01 | 富士フイルム株式会社 | 成形材料 |
JPWO2019146557A1 (ja) * | 2018-01-23 | 2020-12-17 | 富士フイルム株式会社 | 成形材料 |
JPWO2019146558A1 (ja) * | 2018-01-23 | 2021-01-07 | 富士フイルム株式会社 | 多孔成形体 |
JP7113034B2 (ja) | 2018-01-23 | 2022-08-04 | 富士フイルム株式会社 | 多孔成形体 |
WO2020204285A1 (ko) * | 2019-04-02 | 2020-10-08 | 한국과학기술원 | 3차원 나노구조의 고정상을 갖는 가스 크로마토그래피용 마이크로 분별기 및 그 제조 방법 |
US12055528B2 (en) | 2019-04-02 | 2024-08-06 | Korea Advanced Institute Of Science And Technology | Micro-separator having stationary phase with three dimensional nano-structure and method for manufacturing the same |
JP2023531012A (ja) * | 2020-09-15 | 2023-07-20 | エルジー・ケム・リミテッド | 細胞培養用マイクロキャリア、その製造方法およびこれを用いた細胞培養組成物 |
JP7542909B2 (ja) | 2020-09-15 | 2024-09-02 | エルジー・ケム・リミテッド | 細胞培養用マイクロキャリア、その製造方法およびこれを用いた細胞培養組成物 |
Also Published As
Publication number | Publication date |
---|---|
EP2945982A2 (en) | 2015-11-25 |
JP2013504763A (ja) | 2013-02-07 |
JP5789261B2 (ja) | 2015-10-07 |
WO2011032704A2 (en) | 2011-03-24 |
WO2011032703A2 (en) | 2011-03-24 |
WO2011032705A3 (en) | 2011-05-26 |
WO2011032703A3 (en) | 2011-09-15 |
US20120237606A1 (en) | 2012-09-20 |
GB2473814A (en) | 2011-03-30 |
WO2011032705A2 (en) | 2011-03-24 |
CN102630241A (zh) | 2012-08-08 |
GB2473814B (en) | 2014-06-11 |
US8906404B2 (en) | 2014-12-09 |
US20120309053A1 (en) | 2012-12-06 |
GB0916281D0 (en) | 2009-10-28 |
CN102630240A (zh) | 2012-08-08 |
CN102630241B (zh) | 2015-05-27 |
EP2478045A2 (en) | 2012-07-25 |
CN102630240B (zh) | 2015-07-08 |
WO2011032704A3 (en) | 2011-09-22 |
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