System and method for isolating extracellular vesicles
Inventors
Katakowski, Mark • Hozeska-Solgot, Ann
Assignees
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Publication Number
US-11660271-B2
Publication Date
2023-05-30
Expiration Date
Abstract
A system and method of isolating extracellular vesicles. The method includes loading one or more of blood or bone marrow into an input port of a concentration system and centrifuging one or more of the blood or bone marrow to separate one or more of red blood cells, platelet poor plasma, or platelet rich plasma/bone marrow concentrate fractions via a centrifuge device. The method further includes pumping one or more of bone marrow/platelet rich plasma fractions and platelet poor plasma fractions into a receptacle of the concentration system and adding a concentrated aqueous two-phase solution to one or more of the bone marrow concentrate/platelet rich plasma fractions and platelet poor plasma fractions. The method also includes drawing the concentrated aqueous two-phase solution and one or more of the bone marrow concentrate/platelet rich plasma fractions or platelet poor plasma fractions back into the centrifuge device to isolate one or more of extracellular vesicles and platelet rich plasma/bone marrow concentrate fractions.
Core Innovation
The disclosed invention relates to a method of isolating extracellular vesicles or platelet rich plasma fractions from blood or bone marrow using a concentration system having an input port, a centrifuge device, and receptacles. Blood or bone marrow is loaded into the input port and centrifuged to separate red blood cells, platelet poor plasma fractions, or platelet rich plasma fractions, and platelet rich plasma fractions and platelet poor plasma fractions are collected into a first receptacle of the concentration system.
A concentrated aqueous two-phase solution is added to the platelet rich plasma fractions and/or the platelet poor plasma fractions, and the concentrated aqueous two-phase solution and selected platelet fractions are drawn back into the centrifuge device for further centrifugation. The method isolates extracellular vesicles from the platelet poor plasma fractions and isolates extracellular vesicles and platelet rich plasma fractions from the platelet rich plasma fractions, producing a pellet for injection comprising the extracellular vesicles.
In embodiments, the concentrated aqueous two-phase solution is disposed in a syringe or a receptacle, and platelet poor plasma fractions and/or platelet rich plasma fractions are added to the concentrated aqueous two-phase solution prior to centrifuging. In these embodiments, extracellular vesicles and/or platelet rich plasma fractions are isolated from the centrifuging and prepared such that the fractions are suitable for injection, and the concentrated aqueous two-phase solution is specified as PEG-DEX.
Claims Coverage
The document contains three independent claims. Across these independent claims, the inventive coverage centers on a two-stage centrifugation workflow with platelet fraction separation followed by aqueous two-phase partitioning to isolate extracellular vesicles for injection, use of a concentrated aqueous two-phase solution disposed in a syringe or receptacle, and adding platelet fractions to a concentrated aqueous two-phase solution for preparation of injection-suitable fractions.
Two-stage centrifugation with aqueous two-phase partitioning for injection-ready extracellular vesicle pellets
Loading blood or bone marrow into an input port of a concentration system; centrifuging to separate red blood cells, platelet poor plasma fractions, or platelet rich plasma fractions; collecting platelet rich plasma fractions and platelet poor plasma fractions into a first receptacle; adding a concentrated aqueous two-phase solution to one or more of the platelet rich plasma fractions and the platelet poor plasma fractions; drawing the concentrated aqueous two-phase solution and one or more of the platelet fractions back into the centrifuge device; isolating extracellular vesicles from the platelet poor plasma fractions and extracellular vesicles and platelet rich plasma fractions from the platelet rich plasma fractions, creating a pellet for injection comprising the extracellular vesicles.
Syringe or receptacle disposal of concentrated aqueous two-phase solution and centrifugation to isolate injection-suitable fractions
Disposing a concentrated aqueous two-phase solution in a syringe or a receptacle; adding one or more of platelet poor plasma fractions or platelet rich plasma fractions into the concentrated aqueous two-phase solution; centrifuging the concentrated aqueous two-phase solution and the added platelet fractions to isolate one or more of extracellular vesicles and platelet rich plasma fractions; and preparing one or more of extracellular vesicles or platelet rich plasma fractions such that the fractions are suitable for injection.
Adding platelet fractions to concentrated aqueous two-phase solution, centrifuging, and preparing for injection
Adding one or more of platelet poor plasma fractions or platelet rich plasma fractions to a concentrated aqueous two-phase solution; centrifuging the concentrated aqueous two-phase solution and the platelet fractions to isolate one or more of extracellular vesicles or platelet rich plasma fractions; and preparing for injection one or more of extracellular vesicles or platelet rich plasma fractions from the centrifuging.
Across the independent claims, the inventive coverage focuses on combining platelet fraction separation by centrifugation with a concentrated aqueous two-phase solution to isolate extracellular vesicles and/or platelet rich plasma fractions, and preparing the resulting fractions for injection. The claims differ mainly in whether the workflow is implemented within a concentration system with receptacles and a centrifuge device, or with the concentrated aqueous two-phase solution disposed in a syringe or receptacle before centrifugation.
Stated Advantages
Clinical-timing compatibility.
Use of typically unused platelet poor plasma fraction.
Reduced contamination/error via integrated syringe loading.
Only one centrifuge device despite two centrifugation cycles.
Documented Applications
Preparing extracellular vesicles and/or platelet rich plasma fractions for injection from blood or bone marrow using a concentration system and a concentrated aqueous two-phase solution.
Preparing injection-suitable fractions from extracellular vesicles and/or platelet rich plasma fractions by disposing the concentrated aqueous two-phase solution in a syringe or a receptacle and centrifuging.
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