Device and method of isolating extracellular vesicles

A method of isolating extracellular vesicles from a fluid, the method comprising:

• transferring a first volume of plasma from a transfer device to a container of a centrifugal device, maintaining a second volume of plasma in the transfer device;
• adding an aqueous two-phase solution to the container of the centrifugal device;
• centrifuging the aqueous two-phase solution and the first volume of plasma in the container to form a fraction of extracellular vesicles and platelet-rich plasma;
• inverting the container of the centrifugal device and removing remaining aqueous two-phase solution and depleted platelet-poor plasma from the container;
• moving the container from an inverted position back to an upright position and adding the second volume of plasma from the transfer device into the container of the centrifugal device;
• resuspending the fraction of extracellular vesicles and platelet-rich plasma with the second volume of plasma in the container by one or more of shaking, inverting, vortexing, and/or centrifuging the container;
• and
• inverting the container and removing platelet-rich plasma with extracellular vesicles for placement in an injection device, the volume of platelet-rich plasma and extracellular vesicles equal to the second volume of plasma in the transfer device.

A method of isolating extracellular vesicles from a fluid, the method comprising:

• transferring a first volume of plasma from a transfer device to a container of a centrifugal device;
• adding an aqueous two-phase solution to the container;
• centrifuging the aqueous two-phase solution and the first volume of plasma in the container to form a fraction of extracellular vesicles and platelet-rich plasma;
• inverting the container of the centrifugal device and removing remaining aqueous two-phase solution and depleted platelet-poor plasma from the container;
• moving the container to an upright position and adding a second volume of plasma from the transfer device into the container;
• resuspending the fraction of extracellular vesicles and platelet-rich plasma with the second volume of plasma in the container;
• and
• inverting the container and removing platelet-rich plasma with extracellular vesicles for placement in an injection device.