Methods for separation of particles

• US 6,051,146 A
• Filed: 01/20/1998
• Issued: 04/18/2000
• Est. Priority Date: 01/20/1998
• Status: Expired due to Term

• Current Assignee: Terumo BCT, Inc. (Terumo Corporation)
• Original Assignee: COBE Laboratories Incorporated (Terumo Corporation)
• Inventors: Goodrich, Raymond P., Green, Todd Curtis
• Primary Examiner(s): Kim, John
• Application Number:
  US09/009,378
• Time in Patent Office: 819 Days
• Field of Search: 210/782, 210/787, 210/789, 210/786, 210/806, 494/37, 435/2, 604/4, 604/5, 604/6
• US Class Current: 210/782
• CPC Class Codes: A61M 1/0227 and means for securing the filter against damage, e.g. during centrifugation
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: A61M 1/3696 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: A61M 2205/12 with interchangeable cassettes forming partially or totally the fluid circuit
• CPC Class Codes: B04B 2005/0471 with additional elutriation separation of different particles
• CPC Class Codes: B04B 5/0442 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation

Abstract

A system and method are disclosed for separating particles having different sedimentation velocities. The system includes a fluid chamber for separating the particles form one another. Conduits are provided for flowing liquid carrying particles to the fluid chamber and for mixing the liquid with a diluting liquid. Structure is also provided for separating particles from the liquids after the particles are separated in the fluid chamber. The disclosed methods include forming a saturated fluidized bed of first particles to retain second particles in the fluid chamber. A diluting liquid having a density less than that of the particles is mixed with the liquid carrying particles to reduce overall density of substances in the fluid chamber and thereby reduce Coriolis jetting of liquids in the fluid chamber.

Claims

1. A method of separating first particles and second particles having different sedimentation velocities, comprising:

rotating a fluid chamber about an axis of rotation;

passing, into the fluid chamber, a first liquid carrying the first and second particles;

forming, in the fluid chamber, a saturated fluidized bed including the first particles, to thereby retain the second particles in the fluid chamber;

flowing into the fluid chamber a second liquid, the second liquid having a density less than that of the first and second particles so that the second liquid reduces an overall density of substances separated in the fluid chamber and thereby limits Coriolis jetting of at least one of the first and second liquid in the fluid chamber, wherein the flowing of the second liquid includes flowing the second liquid into the fluid chamber when the saturated fluidized bed is in the fluid chamber; and

allowing the first liquid, the second liquid, and at least a portion of the first particles to flow from the fluid chamber.

2. The method of claim 1, further comprising mixing the second liquid and the first liquid carrying the first and second particles.

3. The method of claim 1, further comprising flowing the first liquid, the second liquid, and the portion of first particles from the fluid chamber into a separation vessel, separating the first particles from the first and second liquid in the separation vessel, and collecting at least the first particles in a container.

4. The method of claim 1, wherein the first liquid includes plasma, the first particles include at least one of red blood cells, T cells, and stem cells, and the second particles include tumor cells, and wherein the first particles forming the saturated fluidized bed include at least the red blood cells.

5. The method of claim 4, wherein the second liquid is an electrolyte solution having a pH of from about 7.0 to about 7.8.

6. The method of claim 5, wherein the second liquid is saline solution having a pH of about 7.4.

7. The method of claim 4, further comprising increasing a flow rate of the second liquid into the fluid chamber so that stem cells are flushed from the fluid chamber, the reduced overall density of the substances in the fluid chamber limiting Coriolis jetting of the second liquid.

8. The method of claim 1, further comprising adding, to at least one of the first liquid and the second liquid, a polymer having the chemical formula HO--(CH₂ CH₂ O)_A --(CHCH₃ CH₂ O)_B --(CH₂ CH₂ O)_A --H, wherein A satisfies the expression 0≦

A≦

150 and B satisfies the expression 0≦

B≦

100.

9. The method of claim 8, wherein A satisfies the expression 75≦

A≦

76 and B is 30.

23. The method of claim 1, wherein the liquid carrying first and second particles is a cell suspension and wherein the second particles include tumor cells.

24. The method of claim 1, wherein at least portions of the passing and the flowing are performed simultaneously.

25. The method of claim 2, wherein at least portions of the passing and the flowing are performed simultaneously.

26. The method of claim 1, wherein the flowing further includes flowing the second liquid into the fluid chamber after passing the first liquid into the fluid chamber.

27. The method of claim 1, wherein the flowing further includes flowing the second liquid into the fluid chamber before the saturated fluidized bed is formed in the fluid chamber.

28. The method of claim 1, wherein the flowing further includes flowing the second liquid into the fluid chamber during formation of the saturated fluidized bed in the fluid chamber.

10. A method of separating tumor cells from a cell suspension, comprising:

rotating a fluid chamber about an axis of rotation;

flowing a cell suspension including at least a first liquid, first cells, and tumor cells into the fluid chamber;

forming, in the fluid chamber, a saturated fluidized bed including at least the first cells, the saturated fluidized bed retaining tumor cells in the fluid chamber;

mixing a second liquid and the first liquid, the second liquid having a density less than that of the cell suspension so that the second liquid reduces overall density of substances separated in the fluid chamber and thereby limits Coriolis jetting of at least one of the first liquid and the second liquid in the fluid chamber, wherein the mixing includes mixing the first and second liquids when the saturated fluidized bed is in the fluid chamber; and

allowing at least the first liquid, the second liquid, and at least a portion of the first cells to flow from the fluid chamber.

11. The method of claim 10, wherein the mixing precedes the flowing.

12. The method of claim 10, wherein the cell suspension includes at least plasma and stem cells, and wherein the method further comprises flowing the second liquid, plasma, and stem cells from the fluid chamber into a separation vessel, separating at least the stem cells from second liquid and plasma in the separation vessel, and collecting at least the stem cells in a container.

13. The method of claim 10, wherein the first cells include at least one of red blood cells and T cells, and wherein the forming includes establishing a saturated fluidized bed with at least one of the red blood cells and the T cells.

14. The method of claim 10, wherein the second liquid is an electrolyte solution having a pH of from about 7.0 to about 7.8.

15. The method of claim 14, wherein the second liquid is saline solution having a pH of about 7.4.

16. The method of claim 10, wherein the cell suspension includes at least stem cells, and wherein the method further comprises flushing stem cells from the fluid chamber with the second liquid, the reduced overall density of the substances in the fluid chamber limiting Coriolis jetting of the second liquid.

17. The method of claim 10, wherein the cell suspension comprises blood components, and wherein the method further comprises separating platelet rich plasma and red blood cells from peripheral blood to obtain the blood components.

18. The method of claim 10, wherein the cell suspension comprises blood components, and wherein the method further comprises separating platelet rich plasma and red blood cells from bone marrow blood to obtain the blood components.

29. The method of claim 10, wherein the mixing further includes mixing the first and second liquids before the saturated fluidized bed is formed in the fluid chamber.

30. The method of claim 10, wherein the mixing further includes mixing the first and second liquids during formation of the saturated fluidized bed in the fluid chamber.

19. A method of reducing particle adhesion in a particle separation procedure, comprising:

rotating a fluid chamber about an axis of rotation;

flowing liquid carrying particles into the fluid chamber;

separating particles in the fluid chamber according to differences in sedimentation velocity;

adding, to the liquid, a polymer having the chemical formula HO--(CH₂ CH₂ O)_A --(CHCH₃ CH₂ O)_B --(CH₂ CH₂ O)_A --H, wherein A satisfies the expression 0≦

A≦

150 and B satisfies the expression 0≦

B≦

100, the polymer reducing adherence of at least some of the particles to improve separation of the particles; and

allowing at least some of the particles to flow from the fluid chamber.

20. The method of claim 19, wherein A satisfies the expression 75≦

A≦

76 and B is 30.

21. The method of claim 19, wherein the separating includes forming a saturated fluidized bed of particles in the fluid chamber.

22. The method of claim 19, wherein the separating includes filtering the particles by elutriation.