JPWO2018154788A1 - 体細胞製造システム - Google Patents
体細胞製造システム Download PDFInfo
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- JPWO2018154788A1 JPWO2018154788A1 JP2018565076A JP2018565076A JPWO2018154788A1 JP WO2018154788 A1 JPWO2018154788 A1 JP WO2018154788A1 JP 2018565076 A JP2018565076 A JP 2018565076A JP 2018565076 A JP2018565076 A JP 2018565076A JP WO2018154788 A1 JPWO2018154788 A1 JP WO2018154788A1
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Abstract
Description
臨床用の体細胞は非常に綺麗に保たれたクリーンルームで作製、保存される必要がある。しかし、要求されるレベルの清潔度を維持するコストは非常に高額である。そのため、臨床用の体細胞を作るのにコストがかかり、産業化への大きな障害となっている。
体細胞の樹立から保存までの一連の作業が複雑であり、手作業によるところが多い。また、体細胞の作製は、個人の技量に負っているところがある。そのため、作製者や実験バッチによって、臨床用体細胞の品質のばらつきが生じうる。
クリーンルームの中で特定のドナー以外の他人の細胞とのクロスコンタミネーションを防ぐために、一人の人間のみに由来する臨床用体細胞が所定の時間をかけてクリーンルーム内で作製される。また臨床用体細胞の樹立、品質評価には長時間を要する。しかし、臨床用体細胞は、クリーンルームで一度に一人の人間のためだけに作製されるので、多くの希望者の臨床用体細胞を作製するのに非常に長い時間を要する。
上述したように、現状、臨床用体細胞の作製は手作業によるところが大きい。しかし、臨床用体細胞を作製することができるのに必要な技術を有する技術者は少ない。
上記のように、本発明を実施の形態によって記載したが、この開示の一部をなす記述及び図面はこの発明を限定するものであると理解するべきではない。この開示から当業者には様々な代替実施の形態、実施の形態及び運用技術が明らかになるはずである。例えば、因子導入装置30は、レトロウイルス、レンチウイルス、及びセンダイウイルス等のウイルスベクターや、プラスミドを用いるトランスフェクション、あるいはタンパク質トランスフェクションにより、細胞を誘導してもよい。あるいは、因子導入装置30は、エレクトロポレーションにより細胞を誘導してもよい。また、導入前細胞送液路20、導入細胞送液路31、第1体細胞送液路51、拡大培養送液路71、第2体細胞送液路72、及びパッケージ前細胞流路91はマイクロフルイディクス技術により基板上に設けられていてもよい。この様に、本発明はここでは記載していない様々な実施の形態等を包含するということを理解すべきである。
可溶化基底膜調製品(Matrigel、Corning)でコートされた12ウェルディッシュを用意し、各ウェルに10μmol/Lの濃度でROCK(Rho−associated coiled−coil forming kinase/Rho結合キナーゼ)阻害剤(Selleck)を含むフィーダーフリー培地(mTeSR(登録商標)1、STEMCELL Technologies)を入れた。ROCK阻害剤は、細胞死を抑制する。
可溶化基底膜調製品(Matrigel、Corning)でコートされた12ウェルディッシュを用意し、各ウェルに10μmol/Lの濃度でROCK(Rho−associated coiled−coil forming kinase/Rho結合キナーゼ)阻害剤(Selleck)を含むフィーダーフリー培地(mTeSR(登録商標)1、STEMCELL Technologies)を入れた。ROCK阻害剤は、細胞死を抑制する。
可溶化基底膜調製品(Matrigel、Corning)でコートされた12ウェルディッシュを用意し、各ウェルに10μmol/Lの濃度でROCK(Rho−associated coiled−coil forming kinase/Rho結合キナーゼ)阻害剤(Selleck)を含むフィーダーフリー培地(mTeSR(登録商標)1、STEMCELL Technologies)を入れた。
Claims (21)
- 導入前細胞を含む溶液が通過する導入前細胞送液路と、
前記導入前細胞送液路に接続され、前記導入前細胞に体細胞誘導因子を導入して誘導因子導入細胞を作製する因子導入装置と、
前記誘導因子導入細胞を培養して体細胞を作製する細胞作製装置と、
を備える体細胞製造システム。 - 前記導入前細胞送液路、前記因子導入装置、及び前記細胞作製装置を格納する筐体をさらに備える、請求項1に記載の体細胞製造システム。
- 前記体細胞が、多能性幹細胞を除く、請求項1又は2に記載の体細胞製造システム。
- 前記体細胞が、分化細胞を含む、請求項1又は2に記載の体細胞製造システム。
- 前記体細胞が、体性幹細胞を含む、請求項1又は2に記載の体細胞製造システム。
- 前記体細胞が、神経系細胞を含む、請求項1又は2に記載の体細胞製造システム。
- 前記導入前細胞が、多能性幹細胞を含む、請求項1又は2に記載の体細胞製造システム。
- 前記導入前細胞が、体性幹細胞を含む、請求項1又は2に記載の体細胞製造システム。
- 前記導入前細胞が、分化した体細胞を含む、請求項1又は2に記載の体細胞製造システム。
- 前記細胞作製装置が、
前記因子導入装置で作製された誘導因子導入細胞を培養する体細胞培養装置と、
前記体細胞培養装置で樹立された体細胞を拡大培養する拡大培養装置と、
を備え、
前記体細胞培養装置が、前記誘導因子導入細胞に培地を補給する第1の培地補給装置を備え、
前記拡大培養装置が、前記体細胞に培地を補給する第2の培地補給装置を備える、
請求項1から9のいずれか1項に記載の体細胞製造システム。 - 前記体細胞培養装置が、薬剤耐性因子を導入されなかった細胞を死滅させる薬剤を含む溶液を供給する薬剤補給装置をさらに備える、請求項10に記載の体細胞製造システム。
- 前記因子導入装置が、
前記導入前細胞送液路に接続された因子導入部と、
前記体細胞誘導因子を保存する因子保存部と、
前記因子保存部から前記導入前細胞送液路又は前記因子導入部に前記体細胞誘導因子を流すための因子送液路と、
前記因子送液路内の液体を流すためのポンプと、
を備える、請求項1から11のいずれか1項に記載の体細胞製造システム。 - 前記体細胞誘導因子が、DNA、RNA、又はタンパク質である、請求項12に記載の体細胞製造システム。
- 前記因子導入部において、RNAリポフェクションにより、前記導入前細胞に前記体細胞誘導因子が導入される、請求項12に記載の体細胞製造システム。
- 前記体細胞誘導因子がベクターに組み込まれている、請求項12に記載の体細胞製造システム。
- 前記ベクターがセンダイウイルスベクターである、請求項15に記載の体細胞製造システム。
- 前記細胞作製装置で作製された前記体細胞をパッケージするパッケージ装置をさらに備え、前記筐体が前記パッケージ装置を格納する、請求項1から16のいずれか1項に記載の体細胞製造システム。
- 筒状部材と、
前記筒状部材の内部に配置された液体透過フィルターと、
を備える溶液置換器であって、
前記筒状部材に、
前記液体透過フィルター上に、前記細胞作製装置で作製された前記体細胞を含む溶液を導入するための体細胞導入孔と、
前記液体透過フィルター上に、置換溶液を導入するための置換溶液導入孔と、
前記液体透過フィルター上に、前記体細胞を含む置換溶液を流出するための体細胞流出孔と、
前記液体透過フィルターを透過した溶液が流出する廃液流出孔と、
が設けられている溶液置換器
をさらに備える、請求項1から17のいずれか1項に記載の体細胞製造システム。 - 前記廃液流出孔に接続された廃液送液路をさらに備え、前記体細胞を含む溶液の溶液を廃棄する際に前記廃液送液路における溶液の流動が許容され、前記置換溶液中に前記体細胞を分散させる際に前記廃液送液路における溶液の流動が許容されない、請求項18に記載の体細胞製造システム。
- 前記置換溶液が凍結保存液である、請求項18又は19に記載の体細胞製造システム。
- 血液から前記導入前細胞を分離する分離装置をさらに備え、前記分離装置で分離された前記導入前細胞を含む溶液が前記導入前細胞送液路を通過する、請求項1に記載の体細胞製造システム。
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CN110392733A (zh) | 2019-10-29 |
EP3530727A4 (en) | 2020-07-08 |
EP3530727A1 (en) | 2019-08-28 |
JP6480093B2 (ja) | 2019-03-06 |
CA3053891C (en) | 2023-05-23 |
CA3053891A1 (en) | 2018-08-30 |
WO2018154788A1 (ja) | 2018-08-30 |
US20190382706A1 (en) | 2019-12-19 |
US20230002717A1 (en) | 2023-01-05 |
CN110392733B (zh) | 2021-02-26 |
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