JP2020515287A - 合成組織対照及び合成組織マイクロアレイ対照の細胞収率 - Google Patents
合成組織対照及び合成組織マイクロアレイ対照の細胞収率 Download PDFInfo
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Abstract
Description
3−D合成組織対照(STC)は、規定され制御された条件下で正常細胞とある種のがん細胞を懸濁液中で共培養することにより生成される。本明細書において定義するように、「正常」細胞には、非腫瘍細胞が含まれる。正常細胞は、間質細胞及び他の適切な細胞型から形成され得る。
Claims (20)
- 合成組織対照の共培養細胞の収率の増加方法であって、前記方法が:
流動性ゲルを固化させて固形細胞保持体を形成し;
前記細胞保持体に空洞を形成して共培養細胞を保持することを含み、前記空洞が、複数の共培養細胞の細胞培養バッグを保持するように動作可能であり、前記共培養細胞が、少なくとも1つの細胞培養因子に基づいて共培養した正常細胞及びあるがんタイプのがん細胞を含み、前記少なくとも1つの共培養因子が、共培養する前記がん細胞のタイプ、共培養する前記正常細胞に対する前記がん細胞の比、共培養する前記正常細胞と前記がん細胞の播種濃度、前記正常細胞と前記がん細胞の共培養を促進するために使用する細胞増殖サプリメント、及び前記正常細胞と前記がん細胞の共培養を促進するために使用する前記細胞増殖サプリメントのタイプの濃度を含み;前記方法がまた、
前記複数の共培養細胞の細胞培養バッグのうちの少なくとも2つに由来する共培養細胞を、前記細胞保持体の前記空洞内に堆積させ;ならびに
前記共培養細胞を処理して、前記合成組織対照を形成することを含む、前記増加方法。 - 前記共培養細胞の前記処理中に一定量の色素を前記空洞に加え、前記空洞内に堆積した前記共培養細胞を特定することをさらに含む、請求項1に記載の方法。
- 前記空洞に堆積した前記共培養細胞の数に基づいて、前記空洞に加える前記色素の量を決定することをさらに含む、請求項2に記載の方法。
- 前記細胞保持体に前記空洞を形成することが:
前記流動性ゲルが固化している間に物体を前記流動性ゲルに挿入することを含み、前記物体の一部が前記空洞を画定する形状を有し;及び
前記流動性ゲルが固化して前記空洞を形成した後、前記物体を除去することを含む、請求項1に記載の方法。 - さらに、前記合成組織対照をパラフィンブロックに包埋し、合成組織マイクロアレイを形成することを含む、請求項1に記載の方法。
- 前記空洞上に濾材を加え;及び
前記細胞保持体を病理カセットに移し、前記病理カセットを処理して前記合成組織対照を形成することをさらに含む、請求項1に記載の方法。 - がんの存在の判定に使用するための合成組織対照の形成方法であって、前記方法が:
正常細胞及びあるがんタイプのがん細胞を含む複数の細胞の細胞培養バッグを少なくとも1つの細胞培養因子に基づいて共培養することを含み、前記少なくとも1つの細胞培養因子が、培養する前記がん細胞のタイプ、培養する前記正常細胞に対する前記がん細胞の比率、及び培養する前記正常細胞及び前記がん細胞の播種濃度を含み;前記方法がまた、
前記複数の共培養細胞の細胞培養バッグのうちの少なくとも2つの共培養細胞の細胞培養バッグを細胞保持体の空洞に堆積させ;
前記空洞に堆積した前記少なくとも2つの共培養細胞の細胞培養バッグ由来の共培養細胞を処理し;一定量の色素を前記空洞に加え、前記共培養細胞の前記処理中に前記空洞に堆積した前記共培養細胞を特定し;及び
前記空洞に堆積した前記共培養細胞から前記合成組織対照を形成することを含む、前記形成方法。 - 前記複数の細胞の細胞培養バッグを共培養することが、細胞培養チャンバー内のCO2濃度及び温度を前記少なくとも1つの細胞培養因子に基づいたレベルに維持するように構成された前記細胞培養チャンバー内で、前記正常細胞及び前記がん細胞を共培養することを含む、請求項7に記載の方法。
- 前記複数の細胞の細胞培養バッグを共培養することが、前記細胞培養チャンバーを:
前記合成組織対照を保持し;及び
前記少なくとも1つの細胞培養因子に基づいた速度で前記細胞培養チャンバーを回転させるように動作可能な電動回転装置内に維持することをさらに含む、請求項8に記載の方法。 - 前記複数の細胞の細胞培養バッグを共培養することが、前記共培養細胞を培養して、治療応答の診断マーカーとして、及び予測マーカーとして、IHCにおいて使用するマーカーの発現を提供することをさらに含む、請求項8に記載の方法。
- 前記複数の細胞の細胞培養バッグを共培養することが、前記共培養細胞を培養して、治療応答の診断マーカーとして、または予測マーカーとして、IHCにおいて使用するマーカーの発現を提供することをさらに含む、請求項8に記載の方法。
- 前記複数の細胞の細胞培養バッグを共培養することが、前記共培養細胞を培養して、FISH技術で観察する場合にRNAマーカー及びDNAマーカーの一貫した発現レベルを提供することをさらに含む、請求項8に記載の方法。
- 前記複数の細胞の細胞培養バッグを共培養することが、前記共培養細胞を培養して、CISH技術で観察する場合にRNAマーカー及びDNAマーカーの一貫した発現レベルを提供することをさらに含む、請求項8に記載の方法。
- DNaseを加えて、前記正常細胞及び前記がん細胞の非凝集を促進することをさらに含む、請求項7に記載の方法。
- 非凝集の正常細胞を有する均質なコアを形成し;及び
前記均質なコアに前記がん細胞の非凝集細胞を侵入させることをさらに含む、請求項14に記載の方法。 - フィブロネクチンを加えて、前記正常細胞及び前記がん細胞の非凝集を促進することをさらに含む、請求項7に記載の方法。
- 複数の合成組織対照を含む合成組織マイクロアレイであって、前記複数の合成組織対照の各合成組織対照が:
正常細胞;及び
あるがんタイプのがん細胞を含み、
前記正常細胞及び前記がん細胞を、複数の共培養する正常細胞及びがん細胞の細胞培養バッグのうちの少なくとも2つの細胞培養バッグにおいて共培養し、
前記複数の共培養した正常細胞及び前記がん細胞の細胞培養バッグを、培養する前記がん細胞のタイプ、培養する前記正常細胞に対する前記がん細胞の比率、培養する前記正常細胞及び前記がん細胞の播種濃度、ならびに前記正常細胞及びがん細胞の培養を促進するために使用する細胞増殖サプリメントのタイプを含む、少なくとも1つの細胞培養因子に基づいて共培養する、前記合成組織マイクロアレイ。 - 前記正常細胞及びがん細胞を共培養して、IHCまたはISH染色で使用するマーカーの高発現を提供する、請求項17に記載の合成組織マイクロアレイ。
- 前記正常細胞及びがん細胞を共培養して、IHCまたはISH染色で使用するマーカーの中程度の発現を提供する、請求項17に記載の合成組織マイクロアレイ。
- 前記正常細胞及びがん細胞を共培養して、IHCまたはISH染色で使用するマーカーの低発現を提供する、請求項17に記載の合成組織マイクロアレイ。
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