JP2022009363A - マイクロチャネルを有する臓器模倣装置ならびにその使用および製造方法 - Google Patents
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Abstract
Description
本願は、2008年7月16日に出願した米国特許仮出願第61/081,080号の米国特許法119条(e)に基づく恩典を主張するものである。なおその内容全体は、参照により本明細書に組み入れられるものである。
本発明は、米国国立衛生研究所から授与された、助成番号:NIH R01ES016665-01A1による米国政府の支援によってなされたものである。米国政府は本発明に対して一定の権利を有する。
本開示は、概して、マイクロチャネルを有する臓器模倣装置ならびにその使用および製造方法に関する。
押力、引張力、張力、圧縮力などの機械力は、細胞の成長と挙動の重要な制御因子である。テンセグリティーによって、これら物理的力が、どのように細胞中または組織中に分布しているか、ならびにどのようにおよびどこに影響を及ぼすかを決定する構造が提供される。
本発明のシステムと方法は、一つまたは複数の多孔質膜によって分離された中央マイクロチャネルを有する本体を含んでいる。これらの膜は、中央マイクロチャネルを二つ以上の密接した平行な中央マイクロチャネルに分割するように形成され、そして一つまたは複数の第一流体が、第一中央マイクロチャネルを通じて適用され、かつ一つまたは複数の第二流体が第二以降の中央マイクロチャネルを通じて適用される。これら各多孔質膜の表面は、細胞接着分子でコートして細胞の付着を支持し、各膜の上面と下面での組織化を促進して、前記隣接する平行な流体チャネルの間の多孔質膜によって分離された一つまたは複数の組織-組織の界面を生成させることができる。この膜は、生体細胞全体と同様に、多孔質、可撓性、弾性またはそれら性質を組み合わせた性質を有していてもよく、ガスと小さい化学物質のみを交換できる程度に大きいか、または大きなタンパク質の移行とチャネル経由通過が可能な程度に大きい細孔を備えている。また、流体の圧力、流れの特性およびチャネルの形状を変えて、一方のまたは両方の組織層に所望の流体せん断応力を印加することができる。
[本発明1001]
中央マイクロチャネルを内部に有する本体;および
中央マイクロチャネル内に平面に沿って配置された少なくとも部分的に多孔質の膜
を含む、臓器模倣装置であって、
該膜が、中央マイクロチャネルを分離して第一中央マイクロチャネルと第二中央マイクロチャネルを形成するように構成され、第一流体が第一中央マイクロチャネルを通じて適用され、第二流体が第二中央マイクロチャネルを通じて適用され、該膜が、複数種の生体細胞の接着を支持する少なくとも一種類の付着分子でコートされている、臓器模倣装置。
[本発明1002]
第一と第二の中央マイクロチャネルに隣接して配置された、本体の第一チャンバー壁であって、前記膜が取り付けられている第一チャンバー壁、および
第一チャンバー壁の対向する側で第一と第二の中央マイクロチャネルに隣接する第一動作チャネル
をさらに含む、多孔質の膜が少なくとも部分的に可撓性である装置であって、
第一動作チャネルと中央マイクロチャネルの間に印加された圧力差が、第一チャンバー壁を第一の所望の方向に曲げて、第一と第二の中央マイクロチャネル内で平面に沿って拡張または収縮させる、本発明1001の装置。
[本発明1003]
第一と第二の中央マイクロチャネルに隣接して配置された、本体の第二チャンバー壁であって、前記膜の対向する末端が取り付けられている第二チャンバー壁、および
第二チャンバー壁の対向する側で中央マイクロチャネルに隣接して配置された第二動作チャネルであって、第二動作チャネルと中央マイクロチャネルの間の圧力差が、第二チャンバー壁を第二の所望の方向に曲げて、第一と第二の中央マイクロチャネル内で平面に沿って拡張または収縮させる第二動作チャネル
をさらに含む、本発明1002の装置。
[本発明1004]
膜の少なくとも一つの細孔開口の幅寸法が0.5~20ミクロンである、本発明1001の装置。
[本発明1005]
膜がさらに、中央マイクロチャネル内に配置された第一膜と第二膜を含み、第二膜が第一膜に平行に配向されて、それらの間に第三中央マイクロチャネルを形成する、本発明1001の装置。
[本発明1006]
膜がPDMSを含む、本発明1001の装置。
[本発明1007]
膜が一つまたは複数の細胞層でコートされている装置であって、該一つまたは複数の細胞層が膜の表面に適用されている、本発明1001の装置。
[本発明1008]
膜の一方の面または両面が一つまたは複数の細胞層で処理されている装置であって、該一つまたは複数の細胞層が、後生動物細胞、哺乳動物細胞およびヒト細胞からなる群より選択される細胞を含む、本発明1001の装置。
[本発明1009]
細胞が、上皮細胞、内皮細胞、間葉細胞、筋肉細胞、免疫細胞、神経細胞および造血細胞からなる群より選択される、本発明1008の装置。
[本発明1010]
膜の一方の面が上皮細胞で処理され、かつ膜のもう一方の面が内皮細胞で処理されている、本発明1008の装置。
[本発明1011]
装置本体および膜が生体適合材料または生分解性材料で作製されている、本発明1008の装置。
[本発明1012]
さらに生物に移植されている、本発明1008の装置。
[本発明1013]
生物がヒトである、本発明1012の装置。
[本発明1014]
膜が、インビトロで一つまたは複数の細胞層で処理されている、本発明1008の装置。
[本発明1015]
少なくとも一つの膜が、インビボで一つまたは複数の細胞層で処理されている、本発明1008の装置。
[本発明1016]
膜が、少なくとも一つの細胞層の、膜への付着を促進する生体適合性物質でコートされている、本発明1008の装置。
[本発明1017]
生体適合性物質が、コラーゲン、フィブロネクチン、および/またはラミニンを含む細胞外マトリックスである、本発明1016の装置。
[本発明1018]
生体適合材料が、コラーゲン、ラミニン、プロテオグリカン、ビトロネクチン、フィブロネクチン、ポリ-D-リジンおよび多糖からなる群より選択される、本発明1016の装置。
[本発明1019]
第一流体が白血球細胞を含む、本発明1001の装置。
[本発明1020]
本体を有する臓器模倣装置を選択する工程であって、該本体が、中央マイクロチャネル内に平面に沿って配置されて、中央マイクロチャネルを第一中央マイクロチャネルと第二中央マイクロチャネルに分割する、少なくとも部分的に多孔質の膜を含み、該膜が、複数の生体細胞の接着を支持する少なくとも一種類の付着分子でコートされている、選択する工程;
第一流体を第一中央マイクロチャネルを通じて適用する工程;
第二流体を第二中央マイクロチャネルを通じて適用する工程;および
第一と第二の中央マイクロチャネルの間の膜に関する細胞の挙動をモニターする工程
を含む、方法。
[本発明1021]
以下の工程をさらに含む、膜が少なくとも部分的に弾性で、かつ本体が第一と第二の中央マイクロチャネルに隣接して配置された少なくとも一つの動作チャネルを含む、本発明1020の方法:
中央マイクロチャネルと少なくとも一つの動作チャネルの間の圧力差を調節する工程であって、膜が圧力差に応答して平面に沿って伸張する、工程。
[本発明1022]
圧力差を調節する工程がさらに、
膜の一つまたは複数の面が平面に沿って所望の方向に移動するように圧力差を増大すること、および
膜の一つまたは複数の面が平面に沿って逆の方向に移動するように圧力差を低下させること
を含む、本発明1021の方法。
[本発明1023]
膜の少なくとも一つの細孔開口の幅寸法が0.5~20ミクロンである、本発明1021の方法。
[本発明1024]
膜を一つまたは複数の細胞層で処理する工程をさらに含む方法であって、該一つまたは複数の細胞層が膜の表面に適用されている、本発明1021の方法。
[本発明1025]
一つまたは複数の細胞層を膜の一方の面または両面上に適用する工程をさらに含む方法であって、該一つまたは複数の細胞層が後生動物細胞、哺乳動物細胞およびヒト細胞からなる群より選択される細胞を含む、本発明1022の方法。
[本発明1026]
細胞が、上皮細胞、内皮細胞、間葉細胞、筋肉細胞、免疫細胞、神経細胞および造血細胞からなる群より選択される、本発明1025の方法。
[本発明1027]
膜の一方の面が上皮細胞で処理され、かつ膜のもう一方の面が内皮細胞で処理されている、本発明1025の方法。
[本発明1028]
装置本体および膜が生体適合材料または生分解性材料で作製されている、本発明1025の方法。
[本発明1029]
装置が、さらに生物に移植される、本発明1025の方法。
[本発明1030]
生物がヒトである、本発明1029の方法。
[本発明1031]
膜が、インビトロで一つまたは複数の細胞層でコートされる、本発明1025の方法。
[本発明1032]
少なくとも一つの膜が、インビボで一つまたは複数の細胞層でコートされる、本発明1025の方法。
[本発明1033]
膜が、少なくとも一つの細胞層の、膜への付着を促進する生体適合性物質でコートされる、本発明1025の方法。
[本発明1034]
生体適合性物質が、コラーゲン、フィブロネクチンおよび/またはラミニンを含む細胞外マトリックスである、本発明1033の方法。
[本発明1035]
生体適合材料が、コラーゲン、ラミニン、プロテオグリカン、ビトロネクチン、フィブロネクチン、ポリ-D-リジンおよび多糖からなる群より選択される、本発明1033の方法。
[本発明1036]
第一流体が白血球細胞を含有している、本発明1020の方法。
[本発明1037]
本体を有する装置を選択する工程であって、該本体が、中央マイクロチャネル内に平面に沿って配置され、中央マイクロチャネルを、第一中央マイクロチャネルと第二中央マイクロチャネルに分割する少なくとも部分的に多孔質の膜を含む、選択する工程;
前記膜を、該膜の第一面で少なくとも一つの細胞層と接触させ、かつ該多孔質膜の第二面で少なくとも一つの細胞層と接触させて、それにより少なくとも二つの異なる型の細胞を含む組織構造体を形成させる工程;
少なくとも二つの異なる型の細胞を含む前記組織構造体を、利用可能な細胞培養液内で少なくとも一種類の物質と接触させる工程;
均一または不均一な力をある期間、細胞に印加する工程;ならびに
少なくとも二つの異なる型の細胞を含む前記組織構造体内の細胞の反応を測定して、細胞に対する少なくとも一つの物質の作用を決定する工程
を含む、組織系内の少なくとも一種類の物質の作用を、生理学的または病理学的機械力で決定する方法。
[本発明1038]
利用可能な細胞培養液に白血球細胞が添加されている、本発明1037の方法。
[本発明1039]
均一または不均一な力を、真空を使って印加する、本発明1037の方法。
[本発明1040]
少なくとも二つの異なる型の細胞を含む組織構造体が、多孔質膜の第一面上に肺胞上皮細胞を含み、かつ多孔質膜の第二面上に肺微小血管細胞を含む、本発明1037の方法。
[本発明1041]
前記物質が、ナノ粒子、環境毒素もしくは環境汚染物質、タバコの煙、化粧品に使用される化学物質もしくは粒子、薬物もしくは薬物候補、エアロゾル、花粉を含む天然の粒子、化学兵器、一本鎖もしくは二本鎖の核酸、ウイルス、細菌、および単細胞生物からなる群より選択される、本発明1037の方法。
[本発明1042]
反応の測定を、活性酸素種の発現を測定することによって実施する、本発明1037の方法。
[本発明1043]
反応の測定を、組織染色法を利用して実施する、本発明1037の方法。
[本発明1044]
前記物質の作用を測定する前に、少なくとも二つの異なる型の細胞を含む組織構造体を含む膜の生検材料を採取する工程をさらに含む方法であって、該生検材料が染色されている、本発明1037の方法。
[本発明1045]
反応の測定を、接触している細胞培養液の試料から実施する方法であって、反応の測定を、少なくとも二つの異なる型の細胞を含む膜形状の組織構造体の第一面または第二面または両面と接触している細胞培養液の試料から、および少なくとも二つの異なる型の細胞を含む組織構造体を含む膜の第一面または第二面または両面と接触している細胞培養液の試料から実施する、本発明1037の方法。
[本発明1046]
前記物質の作用と、別の物質の作用または前記物質を含まない対照物の作用とを、類似の並列装置系内で比較する工程をさらに含む、本発明1037の方法。
[本発明1047]
膜を少なくとも二種類の物質と接触させる工程をさらに含む方法であって、まず第一物質を接触させて、少なくとも二つの異なる型の細胞を含む組織構造体に対して影響を及ぼし、一定時間経過後、少なくとも第二物質と接触させて、第一物質の作用を受けた少なくとも二つの異なる型の細胞を含む組織構造体に対する第二物質の作用を試験する、本発明1037の方法。
[本発明1048]
中央マイクロチャネルを有する本体、および
中央マイクロチャネル内の平行な平面に沿って配置された複数の膜
を含む臓器模倣装置であって、
前記複数の膜のうちの少なくとも一つが、少なくとも部分的に多孔質であり、かつ前記複数の膜が、中央マイクロチャネルを複数の中央マイクロチャネルに分割するように構成されている、臓器模倣装置。
本明細書の実施態様において、臓器をシミュレートする装置ならびに該装置の使用および製造方法について説明する。当業者は、以下の記載事項が、説明だけを目的とし、限定を全く意図していないことを理解する。その他の態様は、この開示の恩恵を受けている当業者の念頭に容易に浮かぶ。ここで、添付図面に表されている実施態様の実行について、詳細に説明する。同一または類似の品目を指すために図面および下記説明全体を通じて同じ参照指標を使用する。「一態様」という語句は、一より多くの態様を含み、したがって、簡潔さのために一態様のみに限定されないものとする。
が含まれる。一態様では、幹細胞培養物の幹細胞は、誘導された多能性幹細胞である。
中央マイクロチャネル内に平面に沿って配置された少なくとも部分的に多孔質の膜を含み、前記膜が、中央マイクロチャネルを分離して、第一中央マイクロチャネルと第二中央マイクロチャネルを形成するように形成され、第一流体が第一中央マイクロチャネルを通じて利用され、かつ第二流体が第二中央マイクロチャネルを通じて利用され、前記膜が複数の生体細胞の付着を支持する少なくとも一種類の付着分子でコートされている、臓器模倣装置。
第一と第二の中央マイクロチャネルに隣接して配置された、本体の第一チャンバーの壁であって、上記膜がその上に取り付けられている第一チャンバーの壁、および
上記第一チャンバーの壁の対向する側に、第一と第二の中央マイクロチャネルに隣接する第一動作チャネルであって、この動作チャネルと中央マイクロチャネルの間に印加された圧力差が、第一と第二の中央マイクロチャネル内で、第一チャンバーの壁を第一の所望の方向に曲げて、平面に沿って拡張または収縮させる第一動作チャネル
を含む、[A]の装置。
第二チャンバーの壁の対向する側の中央マイクロチャネルに隣接して配置された第二動作チャネルであって、第二動作チャネルと中央マイクロチャネルの間の圧力差が、第一と第二の中央マイクロチャネル内で、第二チャンバーの壁を第二の所望の方向に曲げて平面に沿って拡張または収縮させる第二動作チャネル
をさらに含む、[A]または[B]の装置。
第一流体を第一中央マイクロチャネルを通じて適用する工程、
第二流体を第二中央マイクロチャネルを通じて適用する工程、および
第一と第二の中央マイクロチャネルの間の膜に対する細胞の挙動をモニターする工程
を含む、方法。
中央マイクロチャネルと少なくとも一つの動作チャネルの間の圧力差を調節し、その結果、膜がその圧力差に反応して平面に沿って伸長する工程
をさらに含む、上記パラグラフのいずれかまたはすべての方法。
膜の一つまたは複数の面が平面に沿って所望の方向に移動するように圧力差を増大する工程、および
膜の一つまたは複数の面が平面に沿って反対の方向に移動するように圧力差を低下させる工程
を含む、上記パラグラフのいずれかまたはすべての方法。
前記膜を、該膜の第一面で少なくとも一つの細胞層と接触させ、該多孔質膜の第二面で少なくとも一つの細胞層と接触させて、少なくとも二つの異なる型の細胞を含む組織構造を作製する工程、
少なくとも二つの異なる型の細胞を含む上記組織構造を、適用可能な細胞培養液中の少なくとも一種類の物質と接触させる工程、
均一または不均一な力を、ある期間、細胞に印加する工程、および
少なくとも二つの異なる型の細胞を含む組織構造体中の細胞の反応を測定して、上記少なくとも一種類の物質の細胞に対する作用を測定する工程
を含む、組織系中の少なくとも一種類の物質の作用を、生理学的または病理学的な機械力で決定する方法。
中央マイクロチャネル内の平行面に沿って配置された複数の膜を含み、
複数の膜の少なくとも一つが少なくとも部分的に多孔質であり、該複数の膜が、中央マイクロチャネルを複数の中央マイクロチャネルに分割するよう形成されている、臓器模倣装置。
Claims (1)
- 中央マイクロチャネルを内部に有する本体;および
中央マイクロチャネル内に平面に沿って配置された少なくとも部分的に多孔質の膜
を含む、臓器模倣装置であって、
該膜が、中央マイクロチャネルを分離して第一中央マイクロチャネルと第二中央マイクロチャネルを形成するように構成され、第一流体が第一中央マイクロチャネルを通じて適用され、第二流体が第二中央マイクロチャネルを通じて適用され、該膜が、複数種の生体細胞の接着を支持する少なくとも一種類の付着分子でコートされている、臓器模倣装置。
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