JP2020512812A - 細胞増殖 - Google Patents
細胞増殖 Download PDFInfo
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- JP2020512812A JP2020512812A JP2019553545A JP2019553545A JP2020512812A JP 2020512812 A JP2020512812 A JP 2020512812A JP 2019553545 A JP2019553545 A JP 2019553545A JP 2019553545 A JP2019553545 A JP 2019553545A JP 2020512812 A JP2020512812 A JP 2020512812A
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Abstract
Description
QIC Inlet ≧ 0.2mL/minの場合、
QIC Circ=(−)l/2*QIC Inlet
及び
QIC Inlet = 0.1mL/minの場合、
QIC Circ=(−)QIC Inlet
QIC Inlet ≧ 0.2mL/minの場合、
QIC Circ=(−)l/2*QIC Inlet
及び
QIC Inlet = 0.1mL/minの場合、
QIC Circ=(−)QIC Inlet
以下の説明は、例えば、実施形態の態様を実装するCES500(例えば、図5A、図5B、図5C)及び/又はCES600(図6)のような細胞増殖システムで使用できるプロトコル/方法/プロセスのいくつかの例を含む。実施例では特定の特徴が説明されているが、そのような実施例は単に例示及び説明の目的で提供されている。例えば、実施例では、T細胞及び/又はTreg細胞の増殖が行われるが、他の及び/又は追加の細胞タイプ及び/又はそれらの組み合わせが他の実施形態で使用されてもよい。特定のパラメータ、機能、及び/又は値が説明されているが(いくつかの実施形態では、例えばQuantum(登録商標)細胞増殖システムのようなCESの使用)、これらのパラメータ、機能、及び/又は値等は、単に例示目的で提供される。本開示は、本明細書で提供される実施例及び/又は特定の詳細に限定されない。
以下の表は、T細胞の増殖のためのプロトコル例を実行する数日間にわたる細胞増殖システムの種々のコンポーネント(例えば、ポンプ、揺動装置、バルブ等)のタスク設定(例えば、流量、角回転、出力等)の例を示している。プロトコルは以下のシーケンスに従う。
Claims (65)
- 細胞増殖システムのバイオリアクターへ細胞を投入するステップと、
前記細胞に対してフィーディングを行うステップと、
を有する、
細胞を増殖する方法であって、
フィーディングを行う前記ステップは、
第1の体積の流体を、第1の流量で、前記バイオリアクターの第1のポートに移動させるステップと、
第2の体積の前記流体を、第2の流量で、前記バイオリアクターの第2のポートに移動させるステップと、
を有し、
前記流体は培地を備え、
前記バイオリアクターは複数の前記細胞を含み、
前記第2の流量で前記流体が前記バイオリアクターへ移動する方向は、前記第1の流量で前記流体が前記バイオリアクターへ移動する方向と反対である、
細胞を増殖する方法。 - 請求項1記載の方法において、前記バイオリアクターは中空糸膜を含む、
方法。 - 請求項1記載の方法において、前記複数の前記細胞は、非接着細胞を含む、
方法。 - 請求項1記載の方法において、前記複数の前記細胞は、T細胞を含む、
方法。 - 請求項1記載の方法において、前記第1のポートと前記第2のポートは、前記バイオリアクターの両端部に配置される、
方法。 - 請求項1記載の方法において、前記バイオリアクターの前記第1のポートは、入口ポートである、
方法。 - 請求項1記載の方法において、前記バイオリアクターの前記第2のポートは、前記バイオリアクターの第1の構成において、出口ポートとして機能する、
方法。 - 請求項7記載の方法において、前記バイオリアクターの前記第2のポートは、前記バイオリアクターの第2の構成において、入口ポートとして機能する、
方法。 - 請求項1記載の方法において、前記流体の前記第1の流量は、前記流体の前記第2の流量と等しい、
方法。 - 請求項1記載の方法において、前記流体の前記第1の流量は、前記流体の前記第2の流量と異なる、
方法。 - 請求項1記載の方法において、該方法はさらに、入口体積の前記流体を、入口流量で、第1流体流路に導入するステップを有する、
方法。 - 請求項11記載の方法において、前記流体の前記第1の流量は、前記流体の前記入口流量の第1のパーセンテージ分である、
方法。 - 請求項12記載の方法において、前記流体の前記第1の流量は、前記流体の前記入口流量の約50パーセントである、
方法。 - 請求項12記載の方法において、前記流体の前記第2の流量は、前記流体の前記入口流量の第2のパーセンテージ分である、
方法。 - 請求項14記載の方法において、前記流体の前記第2の流量は、前記流体の前記入口流量の約50パーセントである、
方法。 - 請求項14記載の方法において、前記第1の流量と前記第2の流量との合計は、前記入口流量と等しい、
方法。 - 請求項1記載の方法において、該方法はさらに、前記細胞を増殖するステップを有する、
方法。 - 第1のポートと第2のポートを備えるバイオリアクターと、
少なくとも両端部を有する第1の流体流路と、
前記第1の流体流路と流体的に関連付けられた流体入口路と、
前記第1の流体流路及び前記バイオリアクターと流体的に関連付けられた第1の流体循環路と、
流体を前記第1の流体流路に循環させるための第1のポンプと、
前記流体を前記第1の流体循環路に循環させるための第2のポンプと、
プロセッサと、
前記プロセッサと通信可能であり、且つ前記プロセッサによって読み出し可能であるメモリと、
を備えた、
細胞増殖システムであって、
前記第1のポートと前記第2のポートは、前記バイオリアクターの両端部に位置付けられ、
前記第1の流体流路の第1の端部は、前記バイオリアクターの前記第1のポートと流体的に関連付けられ、前記第1の流体流路の第2の端部は、前記バイオリアクターの前記第2のポートと流体的に関連付けられ、
前記メモリは一連の命令を有し、前記命令が前記プロセッサによって実行されると、前記プロセッサは、
細胞を前記バイオリアクターへ投入することを指示し、
培地を含む前記流体を前記バイオリアクターにおける前記細胞にフィーディングすることを指示し、
前記細胞にフィーディングすることは、
前記第1のポンプを動作させて、前記流体の第1の部分を、第1の流量で、第1の方向に、移動させることと、
前記第2のポンプを動作させて、前記流体の第2の部分を、第2の流量で、前記第1の方向と反対の第2の方向に、移動させることと、
を含む、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記第2のポンプは、前記流体の前記第2の部分を前記バイオリアクターの前記第2のポートに移動する、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記バイオリアクターは、中空糸膜を有する、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記細胞は、非接着細胞を含む、
細胞増殖システム。 - 請求項21記載のシステムにおいて、前記細胞は、T細胞を含む、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記流体の前記第1の部分は、前記流体の前記第2の部分に等しい、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記流体の前記第1の部分は、前記流体の前記第2の部分と異なる、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記流体の前記第1の流量は、前記流体の前記第2の流量に等しい、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記流体の前記第1の流量は、前記流体の前記第2の流量と異なる、
細胞増殖システム。 - 請求項18記載のシステムにおいて、前記流体の前記第2の流量は、前記流体の前記第1の流量のあるパーセンテージ分である、
細胞増殖システム。 - 細胞増殖システムにおいて細胞を増殖する方法であって、該方法は、
複数の細胞を含む第1の体積の流体を、前記細胞増殖システムの第1の流体流路に投入するステップであって、前記第1の流体流路にはバイオリアクターが流体的に関連付けられているステップと、
培地を含む第2の体積の流体を、前記第1の流体流路の一部に投入して、前記第1の体積の流体を、前記バイオリアクターの第1の部分に位置付けるステップと、
前記複数の細胞をアクチベーターにさらすステップと、
第1の期間の間、第1のプロセスに従って、前記細胞に対してフィーディングを行うステップと、
前記第1の期間の間、前記複数の細胞を増殖するステップと、
第2のプロセスに従って、前記細胞に対してフィーディングを行うステップと、
第2の期間の間、前記複数の細胞を増殖するステップと、
前記第2の期間の間、前記複数の細胞を増殖する前記ステップの後、第3の期間の間、前記複数の細胞を、第1の循環流量で、循環させて、第2の細胞クラスターにおける細胞の数を減らすステップと、
前記細胞を収穫するステップと、
を有する、
方法。 - 請求項28記載の方法において、前記第1の期間の間、前記複数の細胞を増殖する前記ステップの後、前記複数の細胞を、前記第1の循環流量で、循環させて、第1の細胞クラスターにおける細胞の数を減らす、
方法。 - 請求項28記載の方法において、前記第2のプロセスは、
培地を含む第3の体積の流体を、第1の流量で、前記バイオリアクターの第1のポートにポンプで移動するステップと、
第4の体積の流体を、第2の流量で、前記バイオリアクターの第2のポートにポンプで移動するステップと、
を有し、
前記流体を前記第2の流量で前記バイオリアクターに移動する方向は、前記流体を前記第1の流量で前記バイオリアクターに移動する方向と反対である、
方法。 - 請求項28記載の方法において、前記複数の細胞は、非接着細胞を含む、
方法。 - 請求項28記載の方法において、前記複数の細胞は、T細胞を含む、
方法。 - 請求項28記載の方法において、前記複数の細胞は、1又は複数のT細胞亜集団を含む、
方法。 - 請求項33記載の方法において、前記1又は複数のT細胞亜集団は、制御性T細胞(Treg)を含む、
方法。 - 請求項28記載の方法において、前記複数の細胞を含む前記第1の体積の流体は、約1×104細胞数/mLから約1×106細胞数/mLで細胞を含む、
方法。 - 細胞増殖システムにおいて細胞を増殖する方法であって、該方法は、
複数の細胞を含む第1の体積の流体を、前記細胞増殖システムに投入するステップであって、前記細胞増殖システムは細胞成長チャンバーを有するステップと、
培地を含む第2の体積の流体を、第1の流体循環路の一部に投入して、前記第1の体積の流体を、前記細胞成長チャンバーの第1の部分に位置付けるステップと、
前記細胞に対してフィーディングを行うステップと、
前記細胞を増殖するステップと、
を有する、
方法。 - 請求項36記載の方法において、前記細胞増殖システムは、第2の流体循環路を有する、
方法。 - 請求項36記載の方法において、前記第1の流体循環路の流体は、前記細胞成長チャンバーの毛細管内側空間を流れる、
方法。 - 請求項38記載の方法において、前記第2の流体循環路の流体は、前記細胞成長チャンバーの毛細管外側空間を流れる、
方法。 - 請求項36記載の方法において、前記複数の細胞を含む前記第1の体積の流体は、毛細管内側循環ポンプを動作させることなく、投入される、
方法。 - 請求項36記載の方法において、前記第1の体積は、前記第2の体積と同じである、
方法。 - 請求項36記載の方法において、前記第1の体積は、前記第2の体積と異なる、
方法。 - 請求項36記載の方法において、前記第1の体積と前記第2の体積との合計は、前記第1の流体循環路の体積のあるパーセンテージ分と等しい、
方法。 - 請求項43記載の方法において、前記第1の体積と前記第2の体積との前記合計は、前記第1の流体循環路の前記体積の約50パーセントである、
方法。 - 請求項36記載の方法において、前記細胞成長チャンバーの前記第1の部分は、前記細胞成長チャンバーの略中央領域である、
方法。 - 請求項36記載の方法において、前記複数の細胞は、1又は複数のT細胞亜集団を含む、
方法。 - 請求項46記載の方法において、前記1又は複数のT細胞亜集団は、制御性T細胞(Treg)を含む、
方法。 - 請求項36記載の方法において、前記細胞成長チャンバーは、中空糸膜を有する、
方法。 - 請求項36記載の方法において、該方法はさらに、前記細胞をアクチベーターで刺激して、前記細胞の増殖を活性化するステップを含む、
方法。 - 請求項36記載の方法において、前記細胞成長チャンバーの入口ポートと接続部との間の前記第1の流体流路の第1の部分は、第3の体積を有する、
方法。 - 請求項50記載の方法において、前記細胞成長チャンバーの前記毛細管内側空間は、第4の体積を有する、
方法。 - 請求項51記載の方法において、前記第2の体積は、少なくとも前記第3の体積と同じである、
方法。 - 請求項52記載の方法において、前記第2の体積は、前記第3の体積と、前記第1の体積の流体によって占有されない前記第4の体積のあるパーセンテージ分との合計と少なくとも同じである、
方法。 - 請求項53記載の方法において、前記第4の体積の前記あるパーセンテージ分は、約5パーセントから約50パーセントである、
方法。 - 細胞増殖システムにおいて細胞を増殖する方法であって、該方法は、
複数の細胞を含む流体を、前記細胞増殖システムにおける細胞成長チャンバーに投入するステップと、
前記複数の細胞をアクチベーターにさらすステップと、
第1の期間の間、前記複数の細胞を増殖するステップと、
前記第1の期間の間、前記複数の細胞を増殖する前記ステップの後、第2の期間の間、前記複数の細胞を、第1の循環流量で、循環させて、細胞クラスターにおける細胞の数を減らすステップと、
を有する、
方法。 - 請求項55記載の方法において、前記複数の細胞を、前記第1の循環流量で、循環させることによって、前記細胞クラスターにせん断応力を引き起こす、
方法。 - 請求項56記載の方法において、前記せん断応力は、少なくとも前記細胞クラスターの第1の細胞を、前記細胞クラスターから分割分離する、
方法。 - 請求項55記載の方法において、前記細胞クラスターにおける細胞の数を減らすことによって、前記細胞クラスターのサイズは、ある減少されたサイズに減らされる、
方法。 - 請求項58記載の方法において、前記細胞クラスターの前記減少されたサイズは、約25μmから約300μmである、
方法。 - 請求項59記載の方法において、前記細胞クラスターの前記減少されたサイズは、約50μmから約250μmである、
方法。 - 請求項60記載の方法において、前記細胞クラスターの前記減少されたサイズは、約75μmから約200μmである、
方法。 - 請求項58記載の方法において、前記細胞クラスターの前記減少されたサイズは、約200μm未満である、
方法。 - 請求項55記載の方法において、前記複数の細胞は、制御性T細胞(Treg)を含む、
方法。 - 請求項55記載の方法において、前記第2の期間の後、第3の期間の間、前記細胞成長チャンバーに位置付けられていない細胞を、前記細胞成長チャンバーに戻す、
方法。 - 請求項64記載の方法において、
前記第3の期間の後、第4の期間の間、前記複数の細胞を増殖し、
前記第4の期間の間、前記複数の細胞を増殖した後、第5の期間の間、前記複数の細胞を、前記第1の循環流量で、循環させて、第2の細胞クラスターにおける細胞の数を減らし、
前記第5の期間の後、第6の期間の間、前記細胞成長チャンバーに位置付けられていない第2の細胞を、前記細胞成長チャンバーに戻す、
方法。
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