JP2022060542A - 細胞を培養するための容器 - Google Patents
細胞を培養するための容器 Download PDFInfo
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Abstract
Description
本出願は、2016年4月12日に出願された“VESSEL FOR CULTURING CELLS”と題する米国仮出願番号第62/321,691号に基づく米国特許法第119条(e)項の下の利益を主張しており、この仮出願は、その全体が参考として本明細書中に援用される。
種々の側面は、細胞を培養するための容器と、懸濁液中の細胞を撹拌および維持するためのそのような容器を使用する方法とに関する。いくつかの側面は、細胞培養インキュベータ内でのそのような容器を使用することと、そのようなインキュベータを使用するための方法とに関する。
細胞培養容器(例えば、スピナーフラスコおよび磁気スターラ)は、細胞を含む液体を掻き混ぜるために使用され、細胞は、撹拌器の回転を介して、懸濁液中で撹拌され、維持される。細胞培養は、研究および臨床状況の両方において有用な技法である。例えば、哺乳類細胞の培養は、多くの場合、クローン細胞株の確立、組織調製、インビトロ受精調製、または幹細胞の集団の増殖のために実施される。
スピナーフラスコおよび磁気スターラ等の細胞を培養するために使用される現在利用可能な容器は、撹拌器(例えば、それぞれ、インペラまたは磁気スターラバー)を使用して、液体の移動を誘発し、これは、順に、液体中の細胞を移動させる。本回転は、細胞を撹拌し、また、懸濁液中の細胞を維持し得る。いくつかの実施形態では、撹拌器(および流体)の回転は、剪断応力を細胞に印加する。残念ながら、細胞は、剪断応力を受けやすく、その結果、細胞培養の間、溶解し得る。いくつかの実施形態では、より小さい容器(例えば、フラスコおよび/またはビーカ)が、使用され、これは、そうでなければより大きい容器内で使用される不必要な培地体積の無駄を回避し得る。しかしながら、そのような実施形態では、より小さい容器内で培養される細胞は、より大きい容器内より頻繁に溶解を被る。
本願明細書は、例えば、以下の項目も提供する。
(項目1)
細胞を培養するための容器であって、
培養されるべき細胞を含む所定の体積の液体を保持するように配列される、容器本体であって、前記懸濁液中の細胞を維持するために十分な前記液体の体積の移動を生成するように配列される、変形可能部分を有する、容器本体
を備える、容器。
(項目2)
前記容器本体は、基部および側壁を備える、項目1に記載の容器。
(項目3)
前記変形可能部分は、前記基部および側壁のうちの少なくとも1つ上に位置する、項目1に記載の容器。
(項目4)
前記変形可能部分は、前記容器本体と一体的に形成される、項目1に記載の容器。
(項目5)
前記変形可能部分は、前記容器本体に除去可能に取付可能である、項目1に記載の容器。
(項目6)
前記変形可能部分の形状は、円形、楕円形、長方形、三角形、および正方形のうちの1つである、項目3に記載の容器。
(項目7)
前記変形可能部分の変形に応答して、前記液体の体積は、移動される、項目1に記載の容器。
(項目8)
前記変形可能部分は、前記容器本体の内部に向かう方向に変形される、項目1に記載の容器。
(項目9)
第2の変形可能部分をさらに備える、項目1に記載の容器。
(項目10)
前記変形可能部分は、前記容器本体の基部および側壁のうちの一方上に位置し、前記第2の変形可能部分は、前記容器本体の基部および側壁の他方上に位置する、項目9に記載の容器。
(項目11)
前記変形可能部分は、変形のパターンに応答して、前記液体の体積の移動を生成するように配列される、項目1に記載の容器。
(項目12)
前記変形可能部分を変形させ、前記液体の体積の移動を生成するように構成される、1つまたはそれを上回る可動部材を有する、変形プレートと組み合わせられる、項目1に記載の容器。
(項目13)
細胞を培養するための容器であって、
培養されるべき細胞を含む所定の体積の液体を保持するように構成される、容器本体であって、基部および側壁を有し、前記基部の少なくとも一部は、変形可能である、容器本体を備え、
前記基部における変形のパターンに応答して、前記細胞は、前記細胞に印加される剪断応力を最小限にしながら、懸濁液中に維持される、容器。
(項目14)
前記基部の部分全体が、変形可能である、項目13に記載の容器。
(項目15)
前記変形のパターンは、波、円形、および蛇行パターンのうちの1つを含む、項目13に記載の容器。
(項目16)
前記基部における変形のパターンに応答して、前記変形可能部分は、前記容器本体の内部に向かう方向に変形される、項目13に記載の容器。
(項目17)
前記側壁の少なくとも一部は、変形可能である、項目13に記載の容器。
(項目18)
前記容器本体は、円筒形に成形される、項目13に記載の容器。
(項目19)
細胞を培養するためのシステムであって、
培養されるべき細胞を含む所定の体積の液体を保持するように構成される、本体を有する、容器であって、変形可能部分を有する、容器本体と、
前記変形可能部分を変形させ、前記懸濁液中の細胞を維持するために十分な前記液体の体積の移動を生成するように配列される、1つまたはそれを上回る可動部材と、
を備える、システム。
(項目20)
筐体および上部表面を有する、変形プレートをさらに備え、前記可動部材は、前記筐体内に配置される、項目19に記載のシステム。
(項目21)
前記1つまたはそれを上回る可動部材は、前方位置に移動し、前記変形可能部分に接触し、それを変形させる、項目20に記載のシステム。
(項目22)
前記前方位置では、前記1つまたはそれを上回る可動部材は、前記上部表面を通して延在する平面を越えて延在する、項目21に記載のシステム。
(項目23)
コントローラをさらに備え、前記コントローラは、前記1つまたはそれを上回る可動部材に、前記変形可能部分を変形させるように命令する、項目19に記載のシステム。
(項目24)
前記1つまたはそれを上回る可動部材を移動させるように配列される、アクチュエータをさらに備える、項目19に記載のシステム。
(項目25)
前記コントローラは、前記1つまたはそれを上回る可動部材に、所定のシークエンスの変形を実施するように命令する、項目23に記載のシステム。
(項目26)
前記1つまたはそれを上回る可動部材は、前記変形可能部分における変形パターンを作成するように配列される、項目23に記載のシステム。
(項目27)
前記変形パターンは、波、円形、および蛇行パターンのうちの1つを含む、項目26に記載のシステム。
(項目28)
細胞を培養する方法であって、
培養されるべき細胞を含む所定の体積の液体を変形可能部分を有する容器内に設置するステップと、
前記懸濁液中の細胞を維持するために十分な前記液体の体積の移動を生産生成するために、前記変形可能部分を変形させるステップと、
を含む、方法。
(項目29)
前記変形可能部分を変形させるステップは、前記変形可能部分を手動で変形させることを含む、項目28に記載の方法。
(項目30)
前記容器を1つまたはそれを上回る可動部材を有する変形プレート上に設置するステップをさらに含み、前記変形可能部分を変形させるステップは、前記1つまたはそれを上回る可動部材を用いて前記変形可能部分を変形させることを含む、項目28に記載の方法。
(項目31)
培養されるべき細胞を含む所定の体積の液体を変形可能部分を有する容器内に設置するステップは、培養されるべき細胞を含む所定の体積の液体を容器本体の側壁および基部のうちの少なくとも1つ上の変形可能部分を有する容器内に設置することを含む、項目28に記載の方法。
(項目32)
前記1つまたはそれを上回る可動部材を用いて前記変形可能部分を変形させることは、前記1つまたはそれを上回る可動部材を前記上部表面を通して延在する平面を越えて延在する前方位置に移動させることを含む、項目30に記載の方法。
(項目33)
前記1つまたはそれを上回る可動部材を用いて前記変形可能部分を変形させることは、前記1つまたはそれを上回る可動部材を用いて前記変形可能部分に接触し、前記変形可能部材を容器本体の内部に向かって移動させることを含む、項目30に記載の方法。
(項目34)
前記1つまたはそれを上回る可動部材を用いて前記変形可能部分を変形させることは、前記変形可能部分における変形パターンを作成することを含む、項目30に記載の方法。
細胞(例えば、スピナーフラスコおよび磁気スターラ)を培養するために使用される現在利用可能な容器は、液体の移動を誘発するために、回転撹拌器に依拠する。例えば、液体は、細胞を撹拌し、懸濁液中の細胞を維持する、インペラまたは磁気スターラを介して回転され得る。いくつかの実施形態では、撹拌器および液体の回転は、剪断応力を細胞に印加する。残念ながら、細胞は、そのような剪断応力を受けやすくあり得、そのような実施形態では、細胞を溶解させ得る。そのような溶解はまた、より大きい培養容器よりもより小さい培養容器において高頻度で見られ得る。そのような実施形態では、細胞溶解は、細胞培養の完全性を損なわせ、結果を役に立たないものにし得る。
自動化された細胞培養
培養容器
初代細胞単離
細胞培養
細胞培養インキュベータ:
材料
監視機器
シール
移送デバイス
コンピュータおよび制御機器
処置および実験介入
本明細書および本請求項で使用されるような、「1つの」という不定冠詞は、明確にそれとは反対に示されない限り、「少なくとも1つの」を意味すると理解されたい。
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