JP7175761B2 - 細胞培養装置、システム及びその使用方法 - Google Patents
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Description
図1は、基部(12)、頂部(14)及び壁要素(8)を含む本発明の折りたたみ可能な細胞培養容器(10)の1つの実施形態を示す。壁要素(8)は、基部(12)と平行して側方に配置された環状剛性部(18、20、22、24、26、28、30)を含む。頂部(14)は、環状剛性部(18)を含み、そして基部(12)は、環状剛性部(30)を含む。環状剛性部及び変形可能領域は、同じか又は異なった直径を有してもよい。この図は、直立構成下での容器を示す。
治療適用のための細胞の処理は、多数の関与する複雑な工程のために複雑であり、且つ労働集約的である。ここで、発明者らは、細胞が単一の容器を出ることなく、細胞治療の生成に関与するすべての工程を実施するために使用され得る新規のプラットフォーム技法を開発し、そして試験して来た。それらの実験においては、折りたたみ可能な円筒状のコンチェルティーナ配置を用いて、容積:表面積比に対する絶妙な制御を可能した。この配置を用いて、細胞を培養し、それらを遠心分離により洗浄し、そして細胞を単一の容器から離れることなく、順番にそれらを凍結融解サイクルに供することが可能であった。
Plp-In(商標)-CHO細胞(Life Technologies)を、10mlの総作用容積中、3×105個の細胞/mlの密度で接種した。それらを、140rpmで作動するシェーキングプラットホーム上で、5%CO2/37℃インキュベーターにて、CDCHO培地(Life Technologies)において増殖した。容器の上部を、0.22μmのMillex GPフィルター(Millipore)に取り付け、無菌性を確保した。3日間のバッチ培養物を毎日サンプリングし、そしてトリパンブルー染色により定量化した(図9)。完全な連続実験(図11(a))のために、細胞をさらに処理した後、24時間さらに増殖した。
遠心分離工程のために、コンチェルティーナを、1000rmpで5分間、遠心分離バケットにおいて回転した。上清液を、コンチェルティーナを反転し、そして圧縮するのみで除去した。上清液を容器から絞り出し、細胞ペレット及び典型的には200-300μlの残留液体を残した。再懸濁のために、10mlの新鮮な培地(又はいくつかの場合、凍結保護剤)を添加する前、コンチェルティーナを直立した位置に戻した。細胞ペレットを除去し、そして単一の細胞懸濁液を、標準的なボルテックスベンチミキサーを用いて得た。細胞を、遠心分離の前、再懸濁時、及び除去された上清液において、細胞数及び生存率について分析した(図10)。
細胞を、10%(v/v)DMSO(Sigma, Poole, UK)を補充されたそれらの正常増殖培地に再懸濁した。装置を、24時間-80℃の冷凍庫に直接配置し、その後、37℃の水浴において融解した。凍結保護剤を、新鮮な培地に細胞を再懸濁する前、上記に概略されるようにしてコンチェルティーナを遠心分離することにより除去した。遠心分離の前後で細胞数を測定した(図11)。
生存細胞濃度及び生存率を、位相差顕微鏡法下で、改良されたNeubauer血球計算器での細胞計数により評価した。100μlの細胞を、水中、0.4%(w/v)トリパンブルー(Sigma, Poole, UK)を用いて1:1に希釈した。1スライド当たり4グリッドからの細胞を計数し、そして細胞死を、10-4mlの単一グリッド体積に基づいて計算した。トリパンブルー排除をまた用いて、膜の完全性に基づいて細胞を区別した。従って、トリパンブルーを排除した細胞は生存可能として採点され、そしてトリパンブルーを排除しなかった細胞は死亡として採点された。生存率は、全細胞集団のパーセンテージとして生存細胞として表わされた。
図9は、折りたたみ可能なコンチェルティーナにおけるCHO細胞の3日間の増殖を示す。増殖曲線は、2日後に対数増殖期に入る前、短い遅滞期を示すそれらのタイプの細胞について典型的である(図9(a))。3日目、生存細胞濃度は、24.8×105個の細胞/mlに達した。3日間で、これは、通常のシェーカーフラスコにおけるそれらのタイプの細胞に典型的な、細胞数の8.2倍の増加を表した。バッチ培養を通して、生存率は85%以上に維持され、このことは、システムが有意なレベルの細胞死を誘発しなかったことを示唆した(図9(b))。
この容器は、細胞が単一の容器を離れることなく、連続した一連の全工程操作のために首尾よく使用された。シェーカーフラスコとして適用した場合、細胞は予測どおりに増殖し、そして同じ装置が、細胞の遠心分離及び再懸濁のため有効であることが判明した。凍結保存工程の間にいくつかの細胞死が観察されたが、しかしこれは、凍結工程の間の温度制御の欠如に起因する可能性がある。これらの実験は、単一の装置が、哺乳類細胞の培養及び保存に関与するすべての工程のための使用され得ることを実証する。
Claims (16)
- 基部、前記基部と平行して配置された頂部、及び前記頂部と前記基部との間に配置され、そして容器の内腔を規定する壁要素を有する細胞培養容器における細胞の培養方法であって、ここで前記容器中の壁要素が前記頂部及び前記基部に対して圧縮可能であり、そして前記容器中の前記頂部が任意に密封可能な入口を有し、前記容器が可撓性材料から構成され、前記細胞培養容器内の培地中において細胞を培養することを含み、
前記方法が、以下:
(iv)細胞の増殖;及び
(v)洗浄;及び
(vi)分離
の工程を含み、
ここで前記方法の各工程は、前記細胞が前記容器の内腔を離れることなく、前記容器の内腔内で実施され、
(i)細胞選択;及び/又は
(ii)遺伝子修飾;及び/又は
(iii)サイトカイン刺激;及び/又は
(vii)前記細胞の凍結保存、
の1又は2以上の工程をさらに任意的に含む、
方法。 - 前記細胞培養容器の壁要素が、前記基部と平行して配置された、壁に複数の側方剛性部を含み、ここで側方剛性部の各対が変形可能領域と交互配置される、請求項1に記載の細胞の培養方法。
- 前記細胞培養容器の壁要素が螺旋コイル領域の何れかの側に提供される変形可能領域を有する剛性螺旋コイル領域を含む、請求項1に記載の細胞の培養方法。
- 前記容器の内腔が複数の連結されたチャンバーを含み、各チャンバーは、対の側方剛性部から形成された一連のセグメントから構成される、請求項1又は2に記載の細胞の培養方法。
- 前記複数の連結されたチャンバーの各々は、複数の連結された各チャンバーの何れかの端に解放可能な閉鎖手段が提供される、請求項1に記載の細胞の培養方法。
- 膜又はフィルターが、対の側方剛性部から形成された複数のセグメントに内腔を分割するために変形可能領域でその内腔内に配置される、請求項1、2又は3のいずれか1項に記載の細胞の培養方法。
- 前記膜又はフィルターが1又は2以上の穴により穿孔される、請求項6に記載の細胞の培養方法。
- 前記膜又はフィルターが、内腔を半分割する、請求項6に記載の細胞の培養方法。
- 前記膜が壁要素と不連続である、請求項6に記載の細胞の培養方法。
- 前記容器の内腔が、前記容器の内腔内に、同心円状に配置された複数の内壁要素を有する、請求項6に記載の細胞の培養方法。
- 前記可撓性材料が、ガス透過性材料である、請求項1~10のいずれか1項に記載の細胞の培養方法。
- 前記可撓性材料が、ポリエチレン(任意には低密度ポリエチレン(LDPE))、シス-1,4-ポリブタジエン、メタクリレート、フタレート、ポリ(塩化ビニリデン)、セルロース、シリコーン、フルオロエチレンポリプロピレン、ポリオレフィン、又はエチレン酢酸ビニルコポリマーから成る群から選択される、請求項1~11のいずれか1項に記載の細胞の培養方法。
- 前記メタクリレートが、ポリ(エチルメタクリレート)を含む、請求項12に記載の細胞の培養方法。
- 前記フタレートが、ポリ(エチレンテレフタレート)を含む、請求項12に記載の細胞の培養方法。
- 前記セルロースが、セルロースアセテートブチレートを含む、請求項12に記載の細胞の培養方法。
- 前記容器が、円形、正方形、長方形、楕円形又は三角形の断面のものである、請求項1~15のいずれか1項に記載の細胞の培養方法。
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Families Citing this family (32)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB201508752D0 (en) * | 2015-05-21 | 2015-07-01 | Mason Christopher And Veraitch Farlan S | Cell culture device, system and methods of use thereof |
US20190264159A1 (en) * | 2016-03-28 | 2019-08-29 | Sony Corporation | Cell culture container, cell culture system, cell culture kit, and cell culture method |
CN109689366B (zh) | 2016-05-05 | 2022-12-02 | 西南研究院 | 用于细胞扩增和相关应用的三维生物反应器 |
KR102538795B1 (ko) * | 2016-11-11 | 2023-05-31 | 오리바이오테크 엘티디 | 세포 배양 장치 시스템 및 그의 사용 방법 |
US11447745B2 (en) | 2017-09-01 | 2022-09-20 | Lonza Walkersville, Inc. | End-to-end cell therapy automation |
US10696961B2 (en) | 2017-12-01 | 2020-06-30 | Global Life Sciences Solutions Usa Llc | Magnetic cell isolation techniques |
US10889792B2 (en) * | 2018-02-09 | 2021-01-12 | Global Life Sciences Solutions Usa Llc | Cell expansion vessel systems and methods |
US11414639B2 (en) | 2018-02-09 | 2022-08-16 | Global Life Sciences Solutions Usa Llc | Bioprocessing vessel |
JP7520322B2 (ja) * | 2018-02-09 | 2024-07-23 | グローバル・ライフ・サイエンシズ・ソリューションズ・ユーエスエー・エルエルシー | 細胞療法のためのバイオプロセッシング方法 |
US11371007B2 (en) | 2018-02-09 | 2022-06-28 | Global Life Sciences Solutions Usa Llc | System and method for fluid flow management in a bioprocessing system |
US11920119B2 (en) | 2018-02-09 | 2024-03-05 | Global Life Sciences Solutions Usa Llc | Systems and methods for bioprocessing |
US11932842B2 (en) | 2018-02-09 | 2024-03-19 | Global Life Sciences Solutions Usa Llc | Bioprocessing apparatus |
JP6870629B2 (ja) * | 2018-02-09 | 2021-05-12 | Jfeエンジニアリング株式会社 | バイオリアクターバッグのスケールアップ方法 |
US11149244B2 (en) | 2018-04-04 | 2021-10-19 | Southwest Research Institute | Three-dimensional bioreactor for T-cell activation and expansion for immunotherapy |
EP3790983B1 (en) * | 2018-05-11 | 2023-07-05 | Exothera SA | Use of flexible or compressible liners in a system and method for clarifying a cell culture harvest solution |
EP3856888A4 (en) | 2018-09-24 | 2022-10-12 | Southwest Research Institute | THREE-DIMENSIONAL BIOREACTORS |
SG11202106384YA (en) | 2018-12-21 | 2021-07-29 | Octane Biotech Inc | Carousel for modular biologic production units |
EP3898994A4 (en) | 2018-12-21 | 2022-12-14 | Lonza Walkersville, Inc. | AUTOMATED VIRAL VECTOR PRODUCTION |
CA3123458A1 (en) | 2018-12-28 | 2020-07-02 | Octane Biotech Inc. | Cell culture and tissue engineering systems with controlled environmental zones |
US12024699B2 (en) * | 2019-01-04 | 2024-07-02 | Oribiotech Ltd. | Systems, devices, and methods for cell processing |
JP2022516324A (ja) * | 2019-01-04 | 2022-02-25 | オリバイオテク・リミテッド | 細胞処理プラットフォーム、細胞処理システム、およびそれを使用する方法 |
CA3128387A1 (en) | 2019-02-08 | 2020-08-13 | Lonza Walkersville, Inc. | Cell concentration methods and devices for use in automated bioreactors |
WO2020176258A1 (en) * | 2019-02-25 | 2020-09-03 | University Of Maryland, Baltimore County | Adjustable fermentation and cell culture flasks |
US20220306993A1 (en) * | 2019-06-10 | 2022-09-29 | I Peace, Inc. | Erythrocyte removal device, mononuclear cell collector, cell culture device, cell culture system, cell culture method, and mononuclear cell collection method |
EP3770600B1 (en) * | 2019-07-25 | 2023-05-10 | CSEM Centre Suisse d'Electronique et de Microtechnique SA - Recherche et Développement | Purification process for cells |
KR20220062576A (ko) * | 2019-09-10 | 2022-05-17 | 이엠디 밀리포어 코포레이션 | 분절형 생물용기 |
GB2592566B (en) | 2020-01-13 | 2022-05-18 | Oribiotech Ltd | An apparatus for, and a method of, processing cells |
KR20220151179A (ko) * | 2020-03-09 | 2022-11-14 | 오리바이오테크 엘티디 | 세포 배양 컨테이너 |
CN111440720A (zh) * | 2020-04-17 | 2020-07-24 | 海口健康岛生物科技有限公司 | 一种免疫细胞培养装置 |
WO2022147196A2 (en) * | 2020-12-31 | 2022-07-07 | Iovance Biotherapeutics, Inc. | Devices and processes for automated production of tumor infiltrating lymphocytes |
GB2609648B (en) * | 2021-08-12 | 2023-11-01 | Oribiotech Ltd | A culture device |
GB2623112A (en) * | 2022-10-07 | 2024-04-10 | Oribiotech Ltd | A device |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005037984A1 (ja) | 2003-10-20 | 2005-04-28 | Jms Co., Ltd. | 細胞取扱装置、組織再生用組成物及び組織再生方法 |
JP2007511205A (ja) | 2003-10-08 | 2007-05-10 | ウィルソン ウォルフ マニュファクチャリング コーポレイション | 気体透過性物質を利用する細胞を培養する方法及び装置 |
Family Cites Families (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1232407A (ja) * | 1968-08-30 | 1971-05-19 | ||
US3712295A (en) * | 1970-08-03 | 1973-01-23 | W Kline | Centrifuge tube |
JPS5287290A (en) * | 1976-01-08 | 1977-07-20 | Kiyouko Takeuchi | Fermentation tank |
US4867172A (en) | 1988-02-23 | 1989-09-19 | Habley Medical Technology Corporation | Collapsible blood collector |
US5399493A (en) * | 1989-06-15 | 1995-03-21 | The Regents Of The University Of Michigan | Methods and compositions for the optimization of human hematopoietic progenitor cell cultures |
US5209372A (en) * | 1992-04-08 | 1993-05-11 | Norwood Peter M | Collapsible spiral container |
JPH0870847A (ja) * | 1994-09-02 | 1996-03-19 | Sumitomo Bakelite Co Ltd | 細胞浮遊培養用容器 |
JP4355212B2 (ja) * | 2001-12-21 | 2009-10-28 | オーガノジェネシス インコーポレーテッド | 細胞培養及び臓器補助用の可変容量チャンバ |
US7033823B2 (en) * | 2002-01-31 | 2006-04-25 | Cesco Bioengineering, Inc. | Cell-cultivating device |
US20070224676A1 (en) * | 2006-03-21 | 2007-09-27 | Becton, Dickinson And Company | Expandable culture roller bottle |
JP5145335B2 (ja) | 2006-06-16 | 2013-02-13 | エクセレレックス インク. | 液体を収容するように形成された容器 |
US8025173B2 (en) | 2006-09-07 | 2011-09-27 | Allegiance Corporation | Collapsible canister liner for medical fluid collection |
WO2009139703A1 (en) * | 2008-05-15 | 2009-11-19 | Ge Healthcare Bio-Sciences Ab | Method for cell expansion |
US8956860B2 (en) * | 2009-12-08 | 2015-02-17 | Juan F. Vera | Methods of cell culture for adoptive cell therapy |
US8641641B2 (en) * | 2010-05-11 | 2014-02-04 | Ethicon Endo-Surgery, Inc. | Instrument for applying therapeutic cells, with proximal portion for processing therapeutic cells |
ES1075366Y (es) * | 2011-08-08 | 2011-12-23 | Gil Mario Beltran | Jeringuilla hipodermica con sistema de fuelle |
US9469671B2 (en) * | 2011-09-03 | 2016-10-18 | Therapeutic Proteins International, LLC | Closed bioreactor |
US9180252B2 (en) * | 2012-04-20 | 2015-11-10 | Bayer Medical Care Inc. | Bellows syringe fluid delivery system |
CA2944220C (en) * | 2013-12-20 | 2024-01-02 | President And Fellows Of Harvard College | Organomimetic devices and methods of use and manufacturing thereof |
US10266940B2 (en) * | 2015-02-23 | 2019-04-23 | Applied Materials, Inc. | Auto capacitance tuner current compensation to control one or more film properties through target life |
GB201508752D0 (en) * | 2015-05-21 | 2015-07-01 | Mason Christopher And Veraitch Farlan S | Cell culture device, system and methods of use thereof |
-
2015
- 2015-05-21 GB GBGB1508752.1A patent/GB201508752D0/en not_active Ceased
-
2016
- 2016-05-20 US US15/575,105 patent/US11208626B2/en active Active
- 2016-05-20 WO PCT/GB2016/051451 patent/WO2016185221A1/en active Application Filing
- 2016-05-20 KR KR1020237020698A patent/KR102658422B1/ko active IP Right Grant
- 2016-05-20 KR KR1020177036831A patent/KR102587068B1/ko active IP Right Grant
- 2016-05-20 IL IL255790A patent/IL255790B2/en unknown
- 2016-05-20 JP JP2018512502A patent/JP7175761B2/ja active Active
- 2016-05-20 EP EP16725209.7A patent/EP3298122A1/en active Pending
-
2018
- 2018-09-28 HK HK18112482.0A patent/HK1253095A1/zh unknown
-
2020
- 2020-12-18 US US17/127,255 patent/US11884911B2/en active Active
-
2021
- 2021-03-24 JP JP2021049742A patent/JP7280907B2/ja active Active
-
2023
- 2023-12-19 US US18/545,965 patent/US20240182844A1/en active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007511205A (ja) | 2003-10-08 | 2007-05-10 | ウィルソン ウォルフ マニュファクチャリング コーポレイション | 気体透過性物質を利用する細胞を培養する方法及び装置 |
WO2005037984A1 (ja) | 2003-10-20 | 2005-04-28 | Jms Co., Ltd. | 細胞取扱装置、組織再生用組成物及び組織再生方法 |
Non-Patent Citations (1)
Title |
---|
2013 Catalogue, Adhesive Dispensing Ltd, 2013年,[検索日 2021.07.06],インターネット<URL:http://pdfs.findtheneedle.co.uk/24913.pdf> |
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