JPWO2015129673A1 - マイクロ流体デバイス及び細胞の微小3次元培養法 - Google Patents
マイクロ流体デバイス及び細胞の微小3次元培養法 Download PDFInfo
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Abstract
Description
(1)マイクロ流体デバイスによる細胞、特にヒトES/iPS細胞等の多能性幹細胞の三次元培養法の開発
(2)ヒドロゲルを用いたヒトES/iPS細胞培養法の三次元化
(3)相転移ヒドロゲルを用いることによる、マイクロ流体デバイスへの細胞の導入・摘出の簡便化、細胞への低ダメージ化
具体的には、本発明は、以下のマイクロ流体デバイス及び微小3 次元培養法を提供するものである。
項1. 少なくとも2つの開口部と接続した細胞培養チャンバーを有し、前記細胞培養チャンバー内に細胞とヒドロゲルを導入して三次元ゲル培地で培養したときに、少なくとも1つの開口部から細胞培養チャンバーへの生理活性物質の供給を前記チャンバー内での濃度勾配を形成しながら行うことができる、マイクロ流体デバイス。
項2. 前記ヒドロゲルは細胞培養チャンバー内で形成される、項1に記載のマイクロ流体デバイス。
項3. 少なくとも1つの開口部と細胞培養チャンバーとを接続する流路が前記チャンバーの径と比較して細いことを特徴とする、項1又は2に記載のマイクロ流体デバイス。
項4. 項1〜3のいずれかに記載のマイクロ流体デバイスの細胞培養チャンバーに細胞と流動状ヒドロゲルを導入する工程、前記ヒドロゲルをゲル状に変換する工程、少なくとも1つの開口部から生理活性物質を細胞培養チャンバー内で濃度勾配を形成するように供給して前記生理活性物質の存在下に細胞を培養する工程を含む、細胞の微小3次元培養法。
項5. 前記細胞が多能性幹細胞である、項4に記載の細胞の微小3次元培養法。
項6. 前記細胞がヒト多能性幹細胞である、項4に記載の細胞の微小3次元培養法。
項7. 複数の生理活性物質を濃度勾配を形成するように細胞培養チャンバーに供給することを特徴とする、項4〜6のいずれかに記載の微小3次元培養法。
この細胞外微小環境は、ヒト多能性幹細胞だけでなく、がん幹細胞においても非常に重要である。
(a)新規3 次元培養用マイクロ流体デバイスの開発
生体内で細胞機能制御に重要な役割を果たしている「細胞外微小環境」は、従来の実験系ではその生体外での再現が非常に困難であった。これは、従来の細胞培養ディッシュなどが大きな(mm〜cm 程度)の空間しか扱えないためであり、細胞外微小環境の再現に必要なμm レベルの実験系では無かった。反面マイクロ流体デバイスは、μm レベルの微小空間を作製することが可能になり、細胞外微小環境の再現に一歩近づく。
(b)相転移ハイドロゲルを用いたヒトES/iPS細胞3次元培養法の開発
ハイドロゲルを用いた細胞培養法では、細胞にダメージを与えること無く、細胞を回収することは非常に困難であった。そこで、本発明の好ましい実施形態ではこの問題を解決するために、可逆的に相転移することができるハイドロゲルを使用した。このハイドロゲルは細胞接着性は低いものの、細胞毒性は無く、3次元培養の際の支持担体として用いることが可能である。また、細胞場用環境下(37℃)ではゲル化し、低温度(20℃)以下では液状になるので、細胞に影響を与えること無く、マイクロ流体デバイスへの導入、摘出が可能になる。
μFDは細胞生物学において様々な利点を持つ反面、作製過程において、鋳型準備に時間がかかることが問題点であった。
(d) マイクロ流体デバイスと組み合わせたハイスループットスクリーニングシステムの開発
多能性幹細胞の機能を維持し、分化を行うために適切な生理活性物質(刺激物)を効率よく同定するために、本発明では、マイクロ流体デバイスと組み合わせたハイスループットスクリーニングシステムを開発した。生理活性物質は、三次元培養において濃度勾配のもとに供給することができ、発生・分化にどのような生理活性物質がどのような濃度勾配で供給されることが適切であるのかについて、本発明のデバイスを用いてハイスループットスクリーニングシステムで決定することができる。
実施例1:3D プリンタを用いたマイクロ流体デバイス作製方法
(1)材料
SYLGARD(登録商標)184 Silicone Elastomer kit (base, curing agent)(Dow corning)
Nunc オムニトレイ(Thermo scientific 165218)
ガラスボトムディッシュ(岩城硝子)
3D プリンタ AGILISTA(Keyence)
デシケーター
コロナフィット CFG-500(信光電気計装株式会社)
3D-CAD(AutoCAD、Blender 等)
(2)操作手順
鋳型の作製
1. 3 次元画像描画ソフトウェア(3D-CAD)により、マイクロ流路構造の型となる鋳型デザインを有するマスクを作製する。
1. Silicone Elastomer Base とCuring agent を10:1 の重量比で撹拌ミキサーを用いて混合する(PDMS混合液)。
1. オムニトレイまたはガラスボトムディッシュをコロナ処理する(コロナフィット CFG-500 等を用いる)。
(1)材料
Mebiol gel メビオール株式会社PMW20-1001 (10ml 希釈用)
mTeSR1
株式会社ベリタス ST-05850
Y-27632
和光純薬株式会社
250-00513 (5mg)
TrypLE Express (1x), Phenol Red Life technologies 12605028 (500ml)
(2)操作手順
Mebiol gel の溶解
Mebiol gel は乾燥状態で重さを計測し、10ml 希釈用に対して6〜10ml のmTeSR1 を加え、4°C で一晩静置して溶解させる。実験の際のMebiol gel 濃度は以下の通り。
Medium 75mg/ml
Hard 91mg/ml
ヒト多能性幹細胞の3次元培養化
1. 流体デバイスはUV 照射15 分間で殺菌
2. Y-27632 を最終濃度10μM で含むmTeSR1 培地を調製
3. MEF 上、またはMatrigel 上の細胞をD-PBS(-)で2 回リンス
4. TrypLE Express を加え37℃で3〜5 分間静置
5. TrypLE Express を吸引除去
6. MEF 上の細胞を用いる場合は、剥離したMEF を除去するため培地で細胞層をリンス
7. mTeSR1+Y-27632 培地で細胞を回収
8. シングルセルにするため、細胞懸濁液をピペッティング
9. 1000rpmで3 分間遠心
10. 上清を吸引除去
11. 細胞ペレットをmTeSR1+Y-27632 培地に再懸濁
12. 細胞数を計測
13. 4×105 cells を新しいチューブに分注
14. 1000rpm 3 分間遠心
15. 上清を吸引除去
(ゲルの凝固防止のため以降の操作は氷上で行い、チップも氷上で冷却した物を用いる)
16. 細胞ペレットを100〜200μl のMebiol gel(mTeSR1 で溶解)に懸濁 (4×105 cells/100〜200μl)
17. この時、最終濃度10μM でY-27632 をMebiol gel に添加
18. 氷上で冷却した流体デバイス内に細胞-Mebiol gel 懸濁液を導入(2〜4×104 cells /10μl)
19. 37℃で5 分間静置してゲル化
20. デバイス上のmedium supplier にmTeSR1+Y-27632 培地を添加
21. 乾燥防止のため、dish に滅菌蒸留水を加える
22. 翌日の培地交換はmTeSR1+Y-27632 培地で行い、それ以降はmTeSR1 培地で毎日1回培地交換を行う。
1. medium supplier 内の古い培地を除去
2. デバイスを氷上に5 分間静置
3. 流体内に新しい培地を加え、ゲルを希釈
4. 流体内から細胞を回収
Claims (7)
- 少なくとも2つの開口部と接続した細胞培養チャンバーを有し、前記細胞培養チャンバー内に細胞とヒドロゲルを導入して三次元ゲル培地で培養したときに、少なくとも1つの開口部から細胞培養チャンバーへの生理活性物質の供給を前記チャンバー内での濃度勾配を形成しながら行うことができる、マイクロ流体デバイス。
- 前記ヒドロゲルは細胞培養チャンバー内で形成される、請求項1に記載のマイクロ流体デバイス。
- 少なくとも1つの開口部と細胞培養チャンバーとを接続する流路が前記チャンバーの径と比較して細いことを特徴とする、請求項1又は2に記載のマイクロ流体デバイス。
- 請求項1〜3のいずれかに記載のマイクロ流体デバイスの細胞培養チャンバーに細胞と流動状ヒドロゲルを導入する工程、前記ヒドロゲルをゲル状に変換する工程、少なくとも1つの開口部から生理活性物質を細胞培養チャンバー内で濃度勾配を形成するように供給して前記生理活性物質の存在下に細胞を培養する工程を含む、細胞の微小3次元培養法。
- 前記細胞が多能性幹細胞である、請求項4に記載の細胞の微小3次元培養法。
- 前記細胞がヒト多能性幹細胞である、請求項4に記載の細胞の微小3次元培養法。
- 複数の生理活性物質を濃度勾配を形成するように細胞培養チャンバーに供給することを特徴とする、請求項4〜6のいずれかに記載の微小3次元培養法。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP3190145B2 (ja) * | 1992-10-30 | 2001-07-23 | 倭 窪田 | 動物組織培養用担体およびこれを用いる動物組織培養方法 |
US20070015137A1 (en) * | 2005-07-05 | 2007-01-18 | Roman Zantl | Microfluid device and method of producing diffusively built gradients |
WO2010056186A1 (en) * | 2008-11-17 | 2010-05-20 | Gradientech Ab | Fluidic culture device |
WO2013151616A1 (en) * | 2012-04-01 | 2013-10-10 | Emd Millipore Corporation | Cell culture and gradient migration assay methods and devices |
WO2014027693A1 (ja) * | 2012-08-16 | 2014-02-20 | 国立大学法人大阪大学 | 被覆細胞の製造方法、及び細胞の三次元構造体の製造方法 |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7374906B2 (en) * | 2000-11-08 | 2008-05-20 | Surface Logix, Inc. | Biological assays using gradients formed in microfluidic systems |
US9637715B2 (en) * | 2005-07-07 | 2017-05-02 | Emd Millipore Corporation | Cell culture and invasion assay method and system |
US20110244567A1 (en) | 2006-08-31 | 2011-10-06 | Snu R&Db Foundation | Device and Method of 3-Dimensionally Generating IN VITRO Blood Vessels |
CA2696339C (en) * | 2007-08-22 | 2017-12-19 | Probiogen Ag | Culture system and method for immunogenicity and immunofunction testing in vitro |
US8921122B2 (en) | 2008-02-11 | 2014-12-30 | The General Hospital Corporation | System and method for quantitative assessment of biological migration behavior |
US9115340B2 (en) * | 2008-08-08 | 2015-08-25 | Agency For Science Technology & Research | Microfluidic continuous flow device |
WO2011014674A2 (en) * | 2009-07-29 | 2011-02-03 | Cornell University | Microfluidic device for pharmacokinetic-pharmacodynamic study of drugs and uses thereof |
US20110186165A1 (en) | 2009-10-05 | 2011-08-04 | Borenstein Jeffrey T | Three-dimensional microfluidic platforms and methods of use and manufacture thereof |
-
2015
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-
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- 2021-08-19 US US17/406,916 patent/US11807844B2/en active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP3190145B2 (ja) * | 1992-10-30 | 2001-07-23 | 倭 窪田 | 動物組織培養用担体およびこれを用いる動物組織培養方法 |
US20070015137A1 (en) * | 2005-07-05 | 2007-01-18 | Roman Zantl | Microfluid device and method of producing diffusively built gradients |
WO2010056186A1 (en) * | 2008-11-17 | 2010-05-20 | Gradientech Ab | Fluidic culture device |
WO2013151616A1 (en) * | 2012-04-01 | 2013-10-10 | Emd Millipore Corporation | Cell culture and gradient migration assay methods and devices |
WO2014027693A1 (ja) * | 2012-08-16 | 2014-02-20 | 国立大学法人大阪大学 | 被覆細胞の製造方法、及び細胞の三次元構造体の製造方法 |
Non-Patent Citations (2)
Title |
---|
CELLDIRECTOR 3D. PRODUCT NOTE. GRADIENTECH AB, 2013, JPN6015019579 * |
Μ-SLIDE VI0.1, INSTRUCTIONS, IBIDI GMBH, 2009, JPN6015034230 * |
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