Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11077230-B2
Publication Date
2021-08-03
Expiration Date
Abstract
The invention provides a method for decellularising a tissue the method comprising the steps of: a) adding a tissue to be decellularised to a decellularisation vessel; b) adding a decellularisation medium comprising at least one detergent and/or at least one enzyme to the vessel; c) introducing a supercritical fluid to the vessel; and d) maintaining contact of the tissue with the supercritical fluid for a time sufficient to substantially decellularise the tissue.
Core Innovation
The invention relates to decellularising tissue by adding a tissue to be decellularised to a decellularisation vessel and adding a decellularisation medium comprising at least one detergent and at least one enzyme selected from the group consisting of a deoxyribonuclease and a ribonuclease. Supercritical carbon dioxide is then introduced to the vessel with the tissue and the decellularisation medium present, and contact is maintained for a time sufficient to substantially decellularise the tissue.
The supercritical-fluid decellularisation uses supercritical CO2, and the decellularisation medium comprises detergents such as SDS, sodium deoxycholate and Triton X-100. The enzyme components include DNase/RNase and proteolytic enzymes such as trypsin, with optional lipase and cellulase.
Supercritical CO2 enables cell disruption and improves penetration and anti-microbial action, while also aiming to preserve tissue and ECM structure. Documented outcomes include reduced DNA while maintaining GAGs and ECM architecture, together with reference to histological and molecular analysis results.
Claims Coverage
The document provides two independent claims, directed to a method of decellularising tissue and a method of manufacturing a tissue implant or scaffold. Each independent claim includes four core inventive steps, with the second claim using the same decellularisation framework for implant or scaffold manufacture.
Decellularisation using supercritical carbon dioxide with detergent and DNase/RNase medium
Adding a tissue to be decellularised to a decellularisation vessel; adding a decellularisation medium comprising at least one detergent and at least one enzyme selected from the group consisting of a deoxyribonuclease and a ribonuclease to the vessel; introducing supercritical carbon dioxide to the vessel with the tissue to be decellularised and the decellularisation medium present; and maintaining contact of the tissue with the supercritical carbon dioxide for a time sufficient to substantially decellularise the tissue.
Manufacturing a tissue implant or scaffold by decellularising with supercritical carbon dioxide and detergent plus DNase/RNase
Adding a tissue to be decellularised to a decellularisation vessel; adding a decellularisation medium comprising at least one detergent and at least one enzyme selected from the group consisting of a deoxyribonuclease and a ribonuclease to the vessel; introducing supercritical carbon dioxide to the vessel with the tissue to be decellularised and the decellularisation medium present; and maintaining contact of the tissue with the supercritical carbon dioxide and the decellularisation medium for a time sufficient to substantially decellularise the tissue.
Across both independent claims, the inventive coverage centers on performing decellularisation in a decellularisation vessel using a decellularisation medium with at least one detergent plus DNase/RNase, combined with introducing supercritical carbon dioxide and maintaining contact for sufficient time to substantially decellularise the tissue. Dependent claims further specify examples of detergents, additional enzyme classes, and a post-decellularisation washing step.
Stated Advantages
Substantially decellularises the tissue while reducing DNA.
Preserves GAGs and ECM architecture.
Improves penetration and provides anti-microbial action.
Documented Applications
Manufacturing a tissue implant or scaffold by decellularising a tissue using the described supercritical carbon dioxide and decellularisation medium.
Interested in licensing this patent?