JP6437598B2 - 組織製品の酵素処理方法 - Google Patents
組織製品の酵素処理方法 Download PDFInfo
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- JP6437598B2 JP6437598B2 JP2017131018A JP2017131018A JP6437598B2 JP 6437598 B2 JP6437598 B2 JP 6437598B2 JP 2017131018 A JP2017131018 A JP 2017131018A JP 2017131018 A JP2017131018 A JP 2017131018A JP 6437598 B2 JP6437598 B2 JP 6437598B2
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- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P41/00—Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Description
「無細胞組織マトリックス」という用語は、本明細書で使用する場合、一般に、細胞および/または細胞成分を実質的に含まない任意の組織マトリックスを指す。皮膚、皮膚の一部(例えば、真皮)、および他の組織、例えば、血管、心臓弁、筋膜、軟骨、骨、神経結合組織などを使用して、本開示の範囲に入る無細胞マトリックスを作製することができる。無細胞組織マトリックスが細胞および/または細胞成分を実質的に含まないかどうかを判定するために、多くの方法で無細胞組織マトリックスの試験または評価を行うことができる。例えば、処理された組織を光学顕微鏡検査で検査して、細胞(生細胞もしくは死細胞)および/または細胞成分が残存するかどうかを判定することができる。さらに、特定のアッセイを使用して細胞または細胞成分の有無を確認することができる。例えば、DNAまたは他の核酸のアッセイを使用して、組織マトリックス中に残存する核物質を定量することができる。一般に、残存するDNAまたは他の核酸がない場合、完全に脱細胞化されている(即ち、細胞および/または細胞成分が除去されている)ことが分かる。最後に、細胞特異的成分(例えば、表面抗原)を同定する他のアッセイを使用して、組織マトリックスが無細胞であるかどうかを判定することができる。皮膚、皮膚の一部(例えば、真皮)、および他の組織、例えば、血管、心臓弁、筋膜、軟骨、骨、および神経結合組織などを使用して、本開示の範囲に入る無細胞マトリックスを作製することができる。
Claims (6)
- 組織マトリックスの処理方法において、
上皮を含まない皮膚組織マトリックスを選択するステップと、
前記皮膚組織マトリックスを第1の脱細胞化溶液で処理して前記皮膚組織マトリックスから細胞および細胞成分の少なくとも一部を除去するステップと、
前記皮膚組織マトリックスに所望のレベルの可撓性を生じさせるのに十分な条件下で、前記皮膚組織マトリックスをタンパク質分解酵素を含む第2の溶液と接触させるステップと、
を含むことを特徴とする方法。 - 請求項1に記載の方法において、前記酵素がブロメラインであることを特徴とする方法。
- 請求項1に記載の方法において、前記酵素がブロメライン、パパイン、フィシン、アクチニジン、またはこれらの組み合わせから選択されることを特徴とする方法。
- 請求項1に記載の方法において、引張強度、引裂強度、縫合強度、耐クリープ性、破裂強度、熱転移温度、またはこれらの組み合わせの少なくとも1つの望ましくない変化を生じさせない条件下で、前記組織マトリックスを前記タンパク質分解酵素と接触させることを特徴とする方法。
- 請求項1に記載の方法において、引張強度、引裂強度、縫合強度、耐クリープ性、熱転移温度、またはこれらの組み合わせの統計学的に有意な低下を引き起こさない条件下で、前記組織マトリックスを前記タンパク質分解酵素と接触させることを特徴とする方法。
- 請求項1に記載の方法において、前記皮膚組織マトリックスから前記細胞および細胞成分の全てを除去するステップを含むことを特徴とする方法。
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