JP6466323B2 - 磁性細胞外マトリックス - Google Patents
磁性細胞外マトリックス Download PDFInfo
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- JP6466323B2 JP6466323B2 JP2015517365A JP2015517365A JP6466323B2 JP 6466323 B2 JP6466323 B2 JP 6466323B2 JP 2015517365 A JP2015517365 A JP 2015517365A JP 2015517365 A JP2015517365 A JP 2015517365A JP 6466323 B2 JP6466323 B2 JP 6466323B2
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Description
本出願は、2012年6月14日出願の米国特許第61/659,970号に対する優先権を主張し、参照により本明細書に組み込まれる。
なし
・ 繊維性(I、II、III、V、XI型)
・ 断続性三重らせんを有する線維付随性(IX、XII、XIV型)
・ 短鎖(VIII、X型)
・ 基底膜(IV型)
・ 他(VI、VII、XIII型)
細胞が磁性ナノ粒子を摂取するような、細胞、細胞の混合物、または組織片のNANOSHUTTLE(商標)、またはその他の磁性ナノ粒子溶液による処理。
細胞が3次元多細胞構造形成から生じるECMを発生させるのを可能にするため、3次元細胞培養が行われる。好ましくは、細胞または細胞の混合物の磁性浮上は、3次元多細胞構造を急速に発生させるその能力、および望ましい大きさに達するまで成長させられるそのような3次元培養によって、行われる。細胞が磁性ナノ粒子を摂取する限り、回転反応器、骨格、懸滴、非粘着性表面、丸底フラスコ、または3次元細胞培養生物反応器などの他の3次元培養方法が、この工程で使用されてもよい。
3次元培養の科学的、酵素的、および/または物理的方法を用いた脱細胞化。
磁性ECMの破片からの磁性分離による、新規脱細胞化された磁性ECMの回復、またはその他の回復方法。
磁性によって保持されるECMの洗浄、またはその他の洗浄方法による、界面活性剤または他の化学物質の除去。
工程1−5は、磁性ナノ粒子ECM上のECM量の強化または増加が望ましいまたは必要な場合には、必要な限り何度繰り返されてもよい。これは、工程5から生じる磁性ECM生産物による工程1の新鮮細胞または組織の再処理によって達成される。
ECMの再播種、および必要に応じたさらなる使用。
Claims (12)
- 磁性ECMを作製する方法であって、
a)細胞が磁性ナノ粒子を摂取するように、前記細胞を前記磁性ナノ粒子を含む組成物と結合させることと、
b)前記細胞が磁化されるのを可能にする期間、前記磁性ナノ粒子を有する前記細胞を磁場において培養することと、
c)前記細胞を、ECMを有する3次元細胞培養に成長させることであり、前記磁性ナノ粒子が前記細胞から失われ、さらに、前記ECMによって保持されるまで成長させることと、
d)細胞片を洗い流しながら前記ECMを保つ磁石を使用して、前記3次元細胞培養を脱細胞化して磁性ECMを作製することと、
を含む、方法。 - 前記組成物が、
i)負電荷を持つナノ粒子と、
ii)正電荷を持つナノ粒子と、
iii)支持分子と、
を含み、
前記負電荷を持つナノ粒子または前記正電荷を持つナノ粒子のうちのどちらかが磁性反応元素を包含し、さらに前記支持分子が前記負電荷を持つナノ粒子および前記正電荷を持つナノ粒子を密接な混合物内で保持する、請求項1に記載の方法。 - a)前記支持分子がペプチド、多糖、核酸、ポリマー、ポリリジン、フィブロネクチン、コラーゲン、ラミニン、BSA、ヒアルロナン、グリコサミノグリカン、アニオン性非硫酸化グリコサミノグリカン、ゼラチン、核酸、細胞外マトリックスタンパク質混合物、抗体、またはこれらの混合体または派生物を含み、b)前記負電荷を持つナノ粒子が金ナノ粒子であり、さらに、c)前記正電荷を持つナノ粒子が酸化鉄ナノ粒子である、請求項2に記載の方法。
- 細胞によって前記磁性ECMを播種することと、前記播種されたECMを磁場で成長させることと、をさらに含む、請求項1に記載の方法。
- 線維芽細胞および/または上皮細胞および/または骨芽細胞、さらに幹細胞および/または前駆細胞によって前記磁性ECMを播種することと、前記播種されたECMを磁場でさらに成長させることと、をさらに含む、請求項1に記載の方法。
- 前記細胞が組織片の形である、請求項1に記載の方法。
- 前記細胞が患者から摘出された組織片の形である、請求項1に記載の方法。
- 患者から採取された幹細胞または前駆細胞によって前記ECMを播種することと、前記播種されたECMを磁場でさらに成長させることと、をさらに含む、請求項7に記載の方法。
- 患者に移植するための組織を作製する方法であって、
a)細胞と、前記細胞によって摂取される磁性ナノ粒子とを結合させることと、
b)前記細胞が融合し浮上するように、前記細胞が磁化されるのを可能にする期間、前記細胞を磁場に曝露することと、
c)前記細胞を、ECMを有する3次元細胞培養を形成するために成長させることであり、前記磁性ナノ粒子が前記細胞から失われ、さらに、前記ECMによって保持されるまで成長させることと、
d)細胞片を洗い流しながら前記ECMを保つ磁石を使用して、前記3次元細胞培養を脱細胞化して磁性ECMを作製することと、
e)前記磁性ECMを患者から採取された同種幹細胞または前駆細胞によって播種することと、
f)前記播種されたECMを磁場で成長させることと、
を含む、方法。 - 前記磁性ナノ粒子が、
i)負電荷を持つナノ粒子と、
ii)正電荷を持つナノ粒子と、
iii)支持分子と、
をさらに含み、
前記負電荷を持つナノ粒子または前記正電荷を持つナノ粒子のうちのどちらかが磁性反応元素を包含し、さらに前記支持分子が前記負電荷を持つナノ粒子および前記正電荷を持つナノ粒子を密接な混合物内で保持する、請求項9に記載の方法。 - 前記磁性ナノ粒子が、吹き付け、注入、エレクトロポレーション、磁気圧力、ヒドロゲル、またはカチオン性リポソームによって提供される、請求項1又は請求項9に記載の方法。
- 磁性ECMの作製方法であって、
a)磁性ナノ粒子を細胞内に設置することと、
b)前記細胞が磁化されるのを可能にする期間、前記磁性ナノ粒子を有する前記細胞を磁場において培養し、さらに、前記磁場で前記細胞を、ECMを有する3次元細胞培養に成長させることであり、前記磁性ナノ粒子が前記細胞から失われ、さらに、前記ECMによって保持されるまで成長させることと、
c)細胞片を洗い流しながら前記ECMを保つ磁石を使用して、前記3次元細胞培養を脱細胞化して磁性ECMを作製することと、
を含む、方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201261659970P | 2012-06-14 | 2012-06-14 | |
US61/659,970 | 2012-06-14 | ||
PCT/US2013/045246 WO2013188449A1 (en) | 2012-06-14 | 2013-06-11 | Magnetic extracellular matrix |
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