Cell Washing Plunger Using Centrifugal Force
US-2016332158-A1 · Nov 17, 2016 · US
US9713810B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9713810-B2 |
| Application number | US-201514672335-A |
| Country | US |
| Kind code | B2 |
| Filing date | Mar 30, 2015 |
| Priority date | Mar 30, 2015 |
| Publication date | Jul 25, 2017 |
| Grant date | Jul 25, 2017 |
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Devices and methods for washing a suspension of cells are provided. The devices include concentric center, intermediate, and outer compartments. The center compartment and the intermediate compartment are in communication with each other by a first valve and the intermediate and outer compartments are in communication with each other by an opening, aperture, or second valve. The methods include loading a suspension of cells into the center compartment, loading a wash solution into the intermediate compartment, and rotating the device. Centripetal force forces cells through the wash solution and into the outer compartment, where they are isolated and collected.
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What is claimed is: 1. A device for washing a suspension of cells, the device comprising: a cylindrical inner wall extending about and along a longitudinal axis from a first end to a second end, the inner wall defining a central compartment; a cylindrical intermediate wall extending about and along the longitudinal axis from the first end to the second end, the cylindrical intermediate wall defining an intermediate compartment adjacent to the central compartment; a cylindrical outer wall extending about and along the longitudinal axis from the first end to a third end, the cylindrical outer wall defining an outer compartment adjacent to the intermediate compartment; a first valve positioned through the inner wall at or near the second end such that the central compartment is in fluid communication with the intermediate compartment when the first valve is open; and an aperture positioned through the intermediate wall at or near the first end such that the intermediate compartment is in fluid communication with the outer compartment. 2. The device according to claim 1 , further comprising a first inlet and a second inlet, the first and second inlets being positioned at the first end, wherein the first and second inlets are in fluid communication with the central and intermediate compartments, respectively. 3. The device according to claim 2 , wherein the first inlet comprises a first vent and the second inlet comprises a second vent, the first and second vents configured to permit air to escape the central compartment and the intermediate compartment, respectively. 4. The device according to claim 1 , further comprising an outlet positioned at the first end, wherein the outlet is in fluid communication with the outer compartment. 5. The device according to claim 4 , wherein the outlet is coupled to a conduit that extends from the outlet to or near the third end. 6. The device according to claim 4 , wherein the outlet comprises an third vent configured to permit air to escape the outer compartment. 7. The device according to claim 1 , further comprising a first vent, a second vent, and a third vent, the vents being positioned on the first end, wherein the first, second, and third vents are in communication with the central, intermediate, and outer compartments, respectively. 8. The device according to claim 1 , wherein the inner wall is sloped outwardly from the first end to the second end such that the central compartment has a diameter D 1 at the first end and a diameter D 2 at the second end, wherein D 1 is less than D 2 . 9. The device according to claim 1 , wherein at least a portion of the intermediate wall is slanted outwardly from or near the second end to the first end such that the outer compartment has a diameter D 3 at the first end and a diameter D 4 at the second end, wherein D 3 is less than D 4 . 10. The device according to claim 1 , wherein the second end and the third end are both at a distance H 1 from the first end. 11. The device according to claim 1 , wherein the inner and intermediate walls extend a distance H 1 from the first end to the second end and the outer wall extends a distance H 2 from the first end to the third end, wherein H 1 is longer than H 2 . 12. The device according to claim 1 , wherein the central, intermediate, and outer compartments are concentric. 13. The device according to claim 1 , wherein the central compartment has a volume equal to or greater than a volume of the outer compartment. 14. The device according to claim 13 , wherein the volume of the central compartment is greater than the volume of the outer compartment. 15. The device according to claim 1 , wherein the first valve is configured to open at a predetermined centripetal force. 16. The device according to claim 1 , wherein the aperture comprises a second valve, such that the intermediate compartment is only in fluid communication with the outer compartment when the second valve is open. 17. The device according to claim 1 , wherein the aperture comprises a second valve configured to open at a predetermined centripetal force. 18. The device according to claim 1 , wherein the first valve comprises a plurality of valves positioned though the inner wall. 19. The device according to claim 1 , wherein the aperture comprises a plurality of second valves positioned through the intermediate wall. 20. The device according to claim 1 , wherein the aperture comprises an annular, continuous opening. 21. The device according to claim 1 , wherein the inner wall is sloped outwardly from the first surface to the second surface at an angle of from about 0.1° to about 5° with respect to the axis. 22. The device according to claim 1 , wherein at least a portion of the intermediate wall is sloped outwardly from or near the second end to the first end at an angle of from about 0.1° to about 5° with respect to the axis. 23. The device according to claim 1 , wherein the first valve is configured to be mechanically opened. 24. A method for washing a suspension of cells, the method comprising: introducing a suspension of cells into a central cylindrical compartment of a device for washing cells; introducing a wash solution into an annular intermediate compartment of the device, wherein the intermediate compartment is adjacent to the central compartment, and wherein the central compartment is separated from the intermediate compartment by a valve; rotating the device about a central longitudinal axis such that the wash solution is forced outward through an aperture and into an annular outer compartment, wherein the outer compartment is adjacent to the intermediate compartment; and opening the valve. 25. The method according to claim 24 , wherein introducing a wash solution to the intermediate compartment comprises introducing a wash solution selected from the group consisting of water, saline, dextrose, saline with 5% dextrose, and phosphate buffered saline into the intermediate compartment. 26. The method according to claim 24 , wherein introducing a suspension of cells into the central compartment comprises introducing a suspension of cells comprising an enhancement composition and a plurality of red blood cells into the central compartment. 27. The method according to claim 26 , further comprising separating the red blood cells from the enhancement composition, wherein separating is accomplished when the plurality of red blood cells are caused to move through the valve and into the intermediate compartment by centripetal force and then through the aperture and into the outer compartment. 28. The method according to claim 27 , further comprising isolating the red blood cells, wherein isolating the red blood cells is accomplished when the red blood cells displace any wash solution in the third compartment such that the third compartment contains only the red blood cells. 29. The method according to claim 28 , further comprising collecting the red blood cells through an outlet in fluid communication with the outer compartment.
with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation · CPC title
for drying or washing the separated solid particles · CPC title
with fluid transport, e.g. in multi-compartment structures · CPC title
with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation · CPC title
Washing or rinsing blood or blood constituents · CPC title
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