Bioengineered allogeneic blood vessel
Inventors
Sumitran-Holgersson, Suchitra • Olausson, Michael
Assignees
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Publication Number
US-12090253-B2
Publication Date
2024-09-17
Expiration Date
Abstract
The present invention relates to methods for recellurization of blood vessels. This method is particularly useful for producing an allogeneic vein, wherein a donor vein is decellularized and then recellularized using whole blood or bone marrow stem cells. The allogeneic veins produced by the methods disclosed herein are particularly advantageous for implantation or transplantation into patients with vascular diseases.
Core Innovation
The disclosed invention relates to decellularizing an allogeneic donor-derived blood vessel by performing at least one treatment cycle in which the blood vessel is perfused with a first solution comprising a detergent and then perfused with a second solution comprising tri-n-butyl phosphate (TnBP). The method expressly avoids perfusing the blood vessel with any DNase solution between the perfusing with the first solution and the second solution, and the decellularizing is characterized as preserving the extracellular matrix of the vessel.
The invention further defines the decellularization sequence as detergent-first followed by tri-n-butyl phosphate, while preserving extracellular matrix components and decellularizing outcomes. Additional refinements include optional DNase-containing perfusion after perfusing with the second solution, including DNase I as the DNase, together with process and quality constraints that the decellularized vessel is free of cell nuclei and HLA class I/II antigens.
The disclosed content also describes subsequent recellularization of the decellularized blood vessel by introducing cells to generate a recellularized vessel. Recellularization is described using endothelial and smooth muscle cells, including CD133+ stem/progenitor cells or whole-blood-derived endothelial/smooth muscle cells, with alternating endothelial and smooth muscle media, and a closed perfusion culture in a bioreactor is described as an optional recellularization approach.
Claims Coverage
The independent claim is directed to a decellularization method using a treatment cycle with a detergent and tri-n-butyl phosphate, with an express DNase-avoidance interval, while preserving extracellular matrix. The dependent claims refine the method by adding optional later DNase perfusion, specific detergent/TnBP identities, and post-decellularization quality constraints including absence of nuclei and HLA class I/II antigens.
Detergent-to-tri-n-butyl phosphate decellularization cycle
A method of decellularizing a blood vessel comprising performing at least one treatment cycle, wherein each of the at least one treatment cycles comprises perfusing the blood vessel to be decellularized with a first solution comprising a detergent, followed by perfusing the blood vessel to be decellularized with a second solution comprising tri-n-butyl phosphate.
DNase avoidance between detergent and tri-n-butyl phosphate
In each treatment cycle, the method comprises perfusing the blood vessel with the first solution comprising a detergent, followed by perfusing with the second solution comprising tri-n-butyl phosphate, wherein the blood vessel is not perfused with a solution comprising DNase between perfusing with the first solution and the second solution.
Extracellular matrix preservation during decellularization
The decellularizing preserves the extracellular matrix of the vessel.
Optional DNase-containing perfusion after tri-n-butyl phosphate
The method further includes, for each treatment cycle, perfusing the blood vessel to be decellularized with a DNase-containing third solution after perfusing with a second solution.
DNase specified as DNase I
The method is carried out using DNase I as the DNase.
Post-decellularization removal of nuclei and HLA class I/II antigens
After performing at least one treatment cycle, the decellularized blood vessel is free of cell nuclei and HLA class I and II antigens.
Across the independent claim and its dependents, the core coverage is the sequential perfusion decellularization using detergent followed by tri-n-butyl phosphate with an explicit restriction against DNase perfusion between those steps, while preserving the extracellular matrix. Dependent refinements add optional later DNase perfusion, specific example detergent/TnBP identities, and acceptance constraints requiring removal of cell nuclei and HLA class I/II antigens.
Stated Advantages
Decellularizing that preserves the extracellular matrix of the vessel.
Decellularized blood vessels that are free of cell nuclei and HLA class I and HLA class II antigens.
Documented Applications
Recellularized decellularized allogeneic blood vessels are described as suitable for patient implantation for vascular disease.
Use of whole-blood-derived endothelial/smooth muscle cells and/or CD133+ stem/progenitor cells for recellularization of decellularized blood vessels is described.
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