JP6612227B2 - バイオリアクターにおける細胞増殖 - Google Patents
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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 (18)
- 細胞増殖システムにおいて接着細胞を投入する方法であって、該方法は、
複数の接着細胞を有する流体を、第1流量で、細胞増殖システムのバイオリアクター内を循環させるステップと、
流体の循環を前記第1流量から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方向に維持して、前記複数の接着細胞のうち少なくとも第1の部分を沈殿させるステップと、
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方向から180度回転した第2方向に回転させるステップと、
前記バイオリアクターを回転させるステップの後、流体を、前記第1流量よりも低い第2流量で、前記バイオリアクター内を循環させるステップと、
前記循環を前記第2流量から減少させるステップと、
第2所定期間の間、前記バイオリアクターを、第2方向に維持して、前記複数の接着細胞のうち少なくとも第2の部分を沈殿させるステップと、
前記バイオリアクターにおいて、前記複数の接着細胞のうち少なくとも前記第1の部分と前記第2の部分と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、該方法はさらに、前記第2所定期間の後、
流体を、前記第2流量よりも低い第3流量で、前記バイオリアクター内を循環させるステップと、
流体の循環レートを前記第3流量から減少させるステップと、
第3所定期間の間、前記バイオリアクターを前記第2方向に維持し、前記複数の接着細胞のうち少なくとも第3の部分を沈殿させるステップと、
を有することを特徴とする方法。 - 請求項2記載の方法において、該方法はさらに、前記第3所定期間の後、
流体を、前記第3流量よりも低い第4流量で、前記バイオリアクター内を循環させるステップと、
流体の循環を前記第4流量から減少させるステップと、
第4所定期間の間、前記バイオリアクターを前記第2方向に維持し、前記複数の接着細胞のうち少なくとも第4の部分を沈殿させるステップと、
を有することを特徴とする方法。 - 請求項3記載の方法において、前記第1所定期間、前記第2所定期間、前記第3所定期間、前記第4所定期間は、同じ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所定期間と前記第2所定期間とは同じ長さ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、10分よりも短いことを特徴とする方法。
- 請求項6記載の方法において、前記第1所定期間は、8分よりも短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1記載の方法において、該方法はさらに、前記第2所定期間の後で且つ前記増殖させるステップの前に、前記バイオリアクターを180度回転して、前記第1方向にするステップを有することを特徴とする方法。
- 接着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、複数の接着細胞を有する流体を、第1流量で、前記バイオリアクター内を循環させるステップと、
前記少なくとも1つのポンプを停止するステップと、
第1所定期間の間、前記バイオリアクターを第1方向に維持して、前記複数の接着細胞のうち少なくとも第1の部分を沈殿させるステップと、
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方向から180度回転した第2方向に回転させるステップと、
前記バイオリアクターを回転させるステップの後、前記少なくとも1つのポンプを作動させて、流体を、前記第1流量よりも低い第2流量で、前記バイオリアクター内を循環させるステップと、
前記少なくとも1つのポンプを停止させるステップと、
第2所定期間の間、前記バイオリアクターを、第2方向に維持して、前記複数の接着細胞のうち少なくとも第2の部分を沈殿させるステップと、
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を介して流体を循環させ、前記バイオリアクターにおいて、前記接着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10記載のシステムにおいて、前記プロセッサ実行可能命令はさらに、
前記第2所定期間の後、
前記少なくとも1つのポンプを作動させて、流体を、前記第2流量よりも低い第3流量で、前記バイオリアクター内を循環させるステップと、
前記少なくとも1つのポンプを停止させるステップと、
第3所定期間の間、前記バイオリアクターを、前記第2方向に維持して、前記複数の接着細胞のうち少なくとも第3の部分を沈殿させるステップと、
を行うための命令を有することを特徴とするシステム。 - 請求項11記載のシステムにおいて、前記プロセッサ実行可能命令はさらに、
前記第3所定期間の後、
前記少なくとも1つのポンプを作動させて、流体を、前記第3流量よりも低い第4流量で、前記バイオリアクター内を循環させるステップと、
前記少なくとも1つのポンプを停止させるステップと、
第4所定期間の間、前記バイオリアクターを、前記第2方向に維持して、前記複数の接着細胞のうち少なくとも第4の部分を沈殿させるステップと、
を行うための命令を有することを特徴とするシステム。 - 請求項12記載のシステムにおいて、前記第1流量は、150ml/分から250ml/分の間にあることを特徴とするシステム。
- 請求項13記載のシステムにおいて、前記第2流量は、50ml/分から150ml/分の間にあることを特徴とするシステム。
- 請求項14記載のシステムにおいて、前記第3流量は、25ml/分から100ml/分の間にあ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、前記第4流量は、50ml/分より小さいことを特徴とするシステム。
- 請求項10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17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。
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JP2016537001A (ja) | 2016-12-01 |
US10557112B2 (en) | 2020-02-11 |
JP2016536998A (ja) | 2016-12-01 |
US10633625B2 (en) | 2020-04-28 |
WO2015073918A1 (en) | 2015-05-21 |
JP6633522B2 (ja) | 2020-01-22 |
CN105793411B (zh) | 2018-04-17 |
US20150140654A1 (en) | 2015-05-21 |
EP3068866A1 (en) | 2016-09-21 |
EP3068866B1 (en) | 2018-04-25 |
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