JP2016536998A - バイオリアクターにおける細胞増殖 - Google Patents
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Abstract
Description
以下では、本発明の実施形態に係るいくつかの例が記載される。しかしながら、以下の実施形態において、例えば、CES(すなわち、QUANTUM(登録商標)細胞増殖システム)のプログラミングを行うために、パラメータ、特徴事項及び/又は値が以下で記載されるが、それらはあくまでも例示のためであり、本発明は、以下で記載されるそれら特定の事項に限定されない。
ここでの目的は、QUANTUM(登録商標)細胞増殖システムにおいて2つの特徴的な細胞播種方法を用いたヒト骨髄由来間葉系幹細胞(hMSCs)の増殖を説明することである。
本発明において中心となる細胞投入プロシージャは、QUANTUM(登録商標)細胞増殖システムのバイオリアクター内において、より均一な細胞分布を可能にすることによって、また播種プロセスにおいて失われる細胞数を減らすことによって、細胞収率を増加するように設計された一連のステップである。
実施例2で記載された同じプロトコルを用いて、本発明の中心となる付着プロトコルの変更例を研究した。上記形態及び上記プロトコルに対する変形(実施形態2)は、循環レート(100ml/分;−50ml/分;25ml/分)後の付着期間の省略である。すなわち、上記した7分間停止の条件の代わりに、停止条件を省略して、次の循環レートが前回の循環レートの直後に続く。対照実験及び実施例2の本発明実験(実施形態1)も比較として行った。
Claims (20)
- 細胞増殖システムにおいて細胞を成長させる方法であって、該方法は、
複数の細胞を有する流体を、細胞増殖システムのバイオリアクター内において循環させるステップと、
流体の循環レートを減少させるステップと、
第1所定期間の間、前記バイオリアクターを、第1水平方向に維持して、前記複数の細胞の少なくとも1部分を沈殿させ、前記バイオリアクターの第1部分に付着させるステップと、
前記第1所定期間の後、前記バイオリアクターを、前記第1水平方向から略180度回転した第2水平方向に回転させるステップと、
前記回転させるステップの後、前記第2水平方向にある前記バイオリアクターにおいて、前記複数の細胞の少なくとも前記第1の部分を増殖させるステップと、
を有することを特徴とする方法。 - 請求項1記載の方法において、前記バイオリアクターは、中空糸膜を備えることを特徴とする方法。
- 請求項2記載の方法において、前記中空糸膜は、複数の中空糸を備えることを特徴とする方法。
- 請求項3記載の方法において、前記バイオリアクターの前記第1部分は、中空糸の少なくとも頂部であることを特徴とする方法。
- 請求項4記載の方法において、前記複数の細胞の少なくとも前記第1の部分を増殖させるステップは、前記複数の細胞の前記第1の部分の成長に重力を影響させて、前記複数の細胞の前記第1の部分を、前記第1部分の下方に位置する前記バイオリアクターの第2部分に向かって成長させることを含むことを特徴とする方法。
- 請求項5記載の方法において、前記バイオリアクターの前記第2部分は、中空糸の少なくとも底部であることを特徴とする方法。
- 請求項1記載の方法において、前記第1所定期間は、前記細胞の前記1部分を付着及び増殖させるに十分な長さの時間であることを特徴とする方法。
- 請求項7記載の方法において、前記第1所定期間は、約10時間よりも長いことを特徴とする方法。
- 請求項1記載の方法において、前記第1所定期間は、約8分よりも短いことを特徴とする方法。
- 請求項9記載の方法において、前記第1所定期間は、約5分であることを特徴とする方法。
- 請求項1記載の方法において、該方法はさらに、前記第1所定期間の後で且つ前記バイオリアクターを第2水平方向に回転させるステップの前に、
前記バイオリアクターを第1鉛直方向に回転させ、前記バイオリアクターを、第2所定期間の間、前記第1鉛直方向に維持するステップと、
前記第2所定期間の後、前記バイオリアクターを、前記第1鉛直方向から略180度回転させた第2鉛直方向に回転させ、前記バイオリアクターを、第3所定期間の間、前記第2鉛直方向に維持するステップと、
を有することを特徴とする方法。 - 請求項11記載の方法において、前記第2所定期間は、前記細胞の前記1部分を増殖させるのに十分な長さであることを特徴とする方法。
- 請求項12記載の方法において、前記第3所定期間は、前記第2所定期間と同じ長さであることを特徴とする方法。
- 請求項12記載の方法において、前記第2所定期間は、少なくとも約10時間であることを特徴とする方法。
- 細胞を増殖させるシステムであって、該システムは、
バイオリアクターと、
前記バイオリアクターに接続され前記バイオリアクターを回転させる少なくとも1つのモータと、
前記バイオリアクターと流体的に結合された少なくとも1つの流体循環路と、
前記少なくとも1つの流体循環路と前記バイオリアクターとにおいて流体を循環させる少なくとも1つのポンプと、
プロセッサ実行可能命令を実行するプロセッサと、
プロセッサ実行可能命令を記憶するメモリと、
を備えたシステムであって、
前記プロセッサ実行可能命令は、前記プロセッサによって実行される場合、
前記少なくとも1つのポンプを作動させて、複数の細胞を有する流体を、前記バイオリアクター内において循環させるステップと、
前記少なくとも1つのポンプの流量を減少させるステップと、
第1所定期間の間、前記バイオリアクターを第1水平方向に維持して、前記複数の細胞の少なくとも1部分を沈殿させ、前記バイオリアクターの第1部分に付着させるステップと、
前記第1所定期間の後、前記少なくとも1つのモータを作動させて、前記バイオリアクターを、前記第1水平方向から略180度回転した第2水平方向に回転させるステップと、
前記回転の後、前記少なくとも1つのポンプを作動させて、前記バイオリアクターにおいて流体を循環させ、前記第2水平方向に位置づけた前記バイオリアクターにおいて、前記細胞を増殖させるステップと、
を有する方法を実行することを特徴とするシステム。 - 請求項15記載のシステムにおいて、前記バイオリアクターは、中空糸膜を備えることを特徴とするシステム。
- 請求項16記載のシステムにおいて、前記中空糸膜は、複数の中空糸を備えることを特徴とするシステム。
- 請求項17記載のシステムにおいて、前記バイオリアクターの前記第1部分は、中空糸の少なくとも頂部であることを特徴とするシステム。
- 請求項18記載のシステムにおいて、前記複数の細胞の前記第1の部分を増殖させるステップは、前記複数の細胞の前記第1の部分の成長に重力を影響させて、前記複数の細胞の前記第1の部分を、前記第1部分の下方に位置する前記バイオリアクターの第2部分に向かって成長させることを含むことを特徴とするシステム。
- 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の前記第2部分は、中空糸の少なくとも底部であることを特徴とするシステム。
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EP3068866A1 (en) | 2016-09-21 |
US10633625B2 (en) | 2020-04-28 |
US20150140653A1 (en) | 2015-05-21 |
CN105793411B (zh) | 2018-04-17 |
US9617506B2 (en) | 2017-04-11 |
JP6612227B2 (ja) | 2019-11-27 |
JP2016537001A (ja) | 2016-12-01 |
EP3068866B1 (en) | 2018-04-25 |
US20150140654A1 (en) | 2015-05-21 |
US20200248126A1 (en) | 2020-08-06 |
US20170275580A1 (en) | 2017-09-28 |
WO2015073918A1 (en) | 2015-05-21 |
US10557112B2 (en) | 2020-02-11 |
CN105992816B (zh) | 2018-04-17 |
US20200255786A1 (en) | 2020-08-13 |
EP3068867B1 (en) | 2018-04-18 |
US11708554B2 (en) | 2023-07-25 |
US11667876B2 (en) | 2023-06-06 |
JP6633522B2 (ja) | 2020-01-22 |
CN105793411A (zh) | 2016-07-20 |
EP3068867A1 (en) | 2016-09-21 |
CN105992816A (zh) | 2016-10-05 |
WO2015073913A1 (en) | 2015-05-21 |
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