JP5731728B2 - 密閉系細胞培養容器及びそれを利用した細胞培養方法 - Google Patents
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Description
また、本発明は、その密閉系細胞培養容器内の細胞培養用空間内のガス透過膜及び/又は物質透過膜表面上で細胞を培養することを特徴とする細胞培養方法を提供する。
さらに、本発明は、密閉系細胞培養容器から得られる培養細胞、並びに培養細胞シートを提供する。
以下に、本発明を実施例に基づいて更に詳しく説明するが、これらは本発明を何ら限定するものではない。
Claims (11)
- 肉厚の弾性材料を利用した止水弁によって密閉された細胞培養用空間を有する、菌に汚染されない密閉系細胞培養容器を用いて細胞を培養することを特徴とする細胞培養方法であって、
細胞培養容器が、(1)細胞培養用空間のガス交換を行うためのガス透過膜が装着され培地の流路が設けられた容器上部部品と(2)細胞培養用表面を有し培地の流路を設けた容器下部部品から構成され、
当該部品の組み合わせにより細胞培養用空間に培地の管状流路が形成され、
当該部品はいずれも切削工程を経ずに各々金型成型により製造され、該容器が該部品を組み合わせてお互いに接合させることにより形成され、該接合の方法が溶着、接着剤による接着、かしめによる接合、およびそれらの組み合わせから選択され、
容器下部部品の細胞培養用表面に0〜80℃の温度範囲で水和力が変化するポリマーが被覆されており、細胞培養用空間内の細胞培養用表面上で細胞を培養することを特徴とする細胞培養方法。 - 肉厚の弾性材料を利用した止水弁によって密閉された細胞培養用空間を有する、菌に汚染されない密閉系細胞培養容器を用いて細胞を培養することを特徴とする細胞培養方法であって、
細胞培養容器が、(1)細胞培養用液体培地のガス交換を行うためのガス透過膜が装着され培地の流路が設けられた容器上部部品と(2)細胞培養用表面となる物質透過膜が装着され培地の流路が設けられた1個以上の容器内部部品と(3)細胞培養用表面を有し培地の流路を設けた容器下部部品から構成され、
上記部品の組み合わせにより細胞培養用空間に培地の管状流路が形成され、
当該部品はいずれも切削工程を経ずに金型成型により製造され、該容器が該部品を組み合わせてお互いに接合させることにより形成され、該接合の方法が溶着、接着剤による接着、かしめによる接合、およびそれらの組み合わせから選択され、
少なくとも容器内部部品の細胞培養用物質透過膜、若しくは容器下部部品の細胞培養用表面の何れかに0〜80℃の温度範囲で水和力が変化するポリマーが被覆されており、細胞培養用空間内の細胞培養用表面上で細胞を培養することを特徴とする細胞培養方法。 - 物質透過膜内の平均孔径が0.2〜3.0μmの範囲である請求項2記載の細胞培養方法。
- 物質透過膜が光学的に透明でその膜上の細胞を顕微鏡で観察できる請求項2または3に記載の細胞培養方法。
- 細胞培養用空間内の複数の細胞培養用表面上で複数の異なる細胞を培養し、下部部品の細胞培養用表面培養される細胞が産生する液性因子だけを物質透過膜を介して混合させながら培養を行うことを特徴とする請求項2〜4のいずれか1項に記載の細胞培養方法。
- ガス透過膜の酸素ガス透過性が0.5×1010cc・cm/cm2・sec・cmHg以上、及び炭酸ガス透過性が3.0×1010cc・cm/cm2・sec・cmHg以上である請求項1〜5のいずれか1項に記載の細胞培養方法。
- ガス透過膜が光学的に透明で細胞培養用表面上の細胞を顕微鏡で観察できる請求項1〜6のいずれか1項に記載の細胞培養方法。
- ガス透過膜の材質がポリカーボネート、ポリスチレン、またはシリコーンポリカーボネートである請求項1〜7のいずれか1項に記載の細胞培養方法。
- 温度応答性ポリマーの被覆量が、0.8〜2.2μg/cm2の範囲である請求項1〜8のいずれか1項に記載の細胞培養方法。
- 0〜80℃の温度範囲で水和力が変化するポリマーがポリ(N−イソプロピルアクリルアミド)である、請求項1〜9のいずれか1項に記載の細胞培養方法。
- 培養細胞を蛋白質分解酵素による処理を行うことなく、温度応答性表面を冷却するだけで剥離することを特徴とする請求項1〜10のいずれか1項に記載の細胞培養方法。
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KR101372966B1 (ko) | 2009-01-23 | 2014-03-13 | 고꾸리쯔 다이가꾸 호우징 오사까 다이가꾸 | 표적 세포 유도용 피더 세포 |
JP5234428B2 (ja) * | 2009-03-30 | 2013-07-10 | 公益財団法人北九州産業学術推進機構 | 細胞集合体形成器具、細胞集合体培養器具、細胞集合体転写キット及び細胞集合体の培養方法 |
WO2012008368A1 (ja) | 2010-07-16 | 2012-01-19 | 株式会社日立製作所 | 細胞培養容器、および細胞培養装置 |
JP5851220B2 (ja) * | 2011-12-02 | 2016-02-03 | 株式会社日立製作所 | 細胞培養容器、その開放器具、及び開放方法 |
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