Methods of harvesting rare cells from blood products

• US 6,544,751 B1
• Filed: 09/30/1999
• Issued: 04/08/2003
• Est. Priority Date: 04/08/1997
• Status: Expired due to Term

• Current Assignee: Pall Corporation (Danaher Corporation)
• Original Assignee: Pall Corporation (Danaher Corporation)
• Inventors: Brandwein, Harvey, Coker, Samuel
• Primary Examiner(s): Smith, Lynette R. F.
• Assistant Examiner(s): PORTNER, VIRGINIA ALLEN
• Application Number:
  US09/402,158
• Time in Patent Office: 1,286 Days
• Field of Search: 204/183.2, 435/7.1, 435/455, 435/325, 435/2, 435/378, 436/519, 436/520, 436/536, 436/537, 210/806, 210/793, 210/678, 424/93.7
• US Class Current: 435/7.1
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system

Abstract

The present invention provides methods of harvesting rare cells from blood products and/or obtaining products of the rare cells. The method includes contacting a blood product containing rare cells with a porous medium, and selectively retaining rare cells of interest on the porous medium. The porous medium can be contacted with an elution fluid wherein a population of the rare cells is eluted from the porous medium. Rare cells selectively retained on the porous medium can be cultured on the porous medium, and products of the rare cells can be obtained.

Claims

1. A method of harvesting rare cells from a blood product comprising:

contacting a blood product containing rare cells with a porous medium comprising a fibrous web;

selectively retaining said rare cells on said porous medium;

contacting said porous medium with a nutrient-rich elution fluid including a viscosity increasing polymer comprising dextran; and

selectively eluting said rare cells from said porous medium.

2. The method of claim 1, wherein said elution fluid further comprises Dulbecco'"'"'s Modified Eagles Medium supplemented with bovine serum albumin and acid citrate-dextrose.

3. The method of claim 1, wherein contacting the porous medium with the elution fluid comprises backflushing the porous medium with the elution fluid.

4. The method of claim 1, wherein said blood product includes at least first and second populations of rare cells, wherein the first population of rare cells is selectively eluted and the second population of rare cells remains selectively retained on the porous medium after contact with said elution fluid.

5. The method of claim 4, wherein said second population consists essentially of dendritic cells, granulocytes, monocytes, macrophages, or stem cells.

6. The method of claim 4, wherein said elution fluid promotes the selective adhesion of said second population of rare cells to said porous medium.

7. The method of claim 4, wherein said elution fluid comprises a divalent cation.

8. The method of The method of claim 4, wherein said elution fluid comprises a cytokine.

9. The method of claim 4, wherein said elution fluid comprises at least one complement complex product.

10. The method of claim 9, wherein said complement complex product is selected from the group consisting of C3a, C4a, and C5a.

11. The method of claim 1, wherein said rare cells comprise lymphocytes.

12. The method of claim 11, further comprising culturing the lymphocytes.

13. The method of claim 1, wherein said rare cells comprise stem cells.

14. The method of claim 1, wherein the rare cells comprise leukocytes.

15. The method of claim 1, further comprising culturing the eluted rare cells.

16. The method of claim 11, further comprising culturing the lymphocytes.

17. The method of claim 14, further comprising culturing the leukocytes.

18. The method of claim 13, further comprising culturing the stem cells.

19. The method of claim 14, wherein the elution fluid further comprises albumin and at least one additional sugar.

20. The method of claim 1, wherein the rare cells are dendritic cells and the method further comprises culturing the dentritic cells.

21. The method of claim 1, wherein the rare cells are mononuclear cells.

22. A method of harvesting rare cells from a blood product comprising:

contacting a blood product containing said rare cells with a porous medium comprising a fibrous web;

selectively retaining said rare cells on said porous medium;

contacting said porous medium with an elution fluid at a temperature of between about 5-37°

C., wherein said elution fluid includes a viscosity increasing polymer comprising dextran; and

selectively eluting said rare cells from said porous medium.

23. The method of claim 22, wherein said elution fluid comprises at least one additional sugar.

24. The method of claim 22, wherein said elution fluid further comprises buffered saline.

25. The method of claim 22, wherein said elution fluid is at a temperature of between about 15-35°

C.

26. The method of claim 25, wherein said elution fluid is at a temperature of between about 20-30°

C.

27. The method of claim 22, wherein the blood product includes at least first and second populations of rare cells, wherein the first population of rare cells is selectively eluted and wherein the second population of rare cells remains selectively retained on the porous medium after contact with said elution fluid.

28. The method of claim 27, wherein said first population consists essentially of lymphocytes.

29. The method of claim 22, wherein said rare cells comprise lymphocytes.

30. The method of claim 22, wherein said rare cells comprise stem cells.

31. The method of claim 22, further comprising culturing the eluted rare cells.

32. The method of claim 29, further comprising culturing the lymphocytes.

33. The method of claim 22, wherein the rare cells comprise leukocytes.

34. The method of claim 33, further comprising culturing the leukocytes.

35. The method of claim 30, further comprising culturing the stem cells.

36. The method of claim 33, wherein the elution fluid further comprises albumin and at least one additional sugar.

37. The method of claim 22, wherein the rare cells are dendritic cells and the method further comprises culturing the dendritic cells.

38. The method of claim 22, wherein the rare cells are mononuclear cells.

39. The method of claim 22, wherein contacting the porous medium with the elution fluid comprises backflushing the porous medium with the elution fluid.

40. A method of harvesting rare cells comprising stem cells from a blood product comprising:

contacting a blood product containing said rare cells comprising stem cells with a porous medium;

selectively retaining said stem cells on said porous medium;

backflushing said porous medium with a nutrient-rich elution fluid at a temperature of between about 5-37°

C., wherein said elution fluid includes a viscosity increasing polymer; and

selectively eluting said stem cells from said porous medium.

41. The method of claim 40, wherein the porous medium comprises a fibrous web.

42. The method of claim 40, further comprising culturing the eluted stem cells.

43. The method of claim 40, further comprising culturing the stem cells.

44. The method of claim 40, wherein the elution fluid further comprises albumin and at least one additional sugar.

45. The method of claim 40, wherein the viscosity increasing polymer comprises dextran.

46. A method of harvesting rare cells from a blood product comprising:

contacting a blood product containing rare cells comprising dendritic cells with a porous medium comprising a fibrous web;

selectively retaining said dendritic cells on said porous medium;

contacting said porous medium with an elution fluid at a temperature of between about 5-37°

C., wherein said elution fluid includes a viscosity increasing polymer;

selectively eluting said dendritic cells from said porous medium; and

culturing the dendritic cells.

47. The method of claim 46, wherein contacting the porous medium with the elution fluid comprises backflushing the porous medium with the elution fluid.

48. A method of harvesting rare cells from a blood product comprising:

contacting a blood product containing rare cells comprising dendritic cells with a porous medium;

selectively retaining said dendritic cells on said porous medium;

contacting said porous medium with a nutrient-rich elution fluid including a viscosity increasing polymer;

selectively eluting said dendritic cells from said porous medium; and

,

culturing the dendritic cells.

49. A method of harvesting rare cells from a blood product comprising:

contacting a blood product including at least first and second populations of rare cells with a porous medium;

selectively retaining said second population of rare cells on said porous medium;

contacting said porous medium with a nutrient-rich elution fluid comprising a viscosity increasing polymer and a cytokine; and

selectively eluting said first population of rare cells from said porous medium.

50. A method of harvesting rare cells from a blood product comprising:

contacting a blood product including at least first and second populations of rare cells with a porous medium;

selectively retaining said second population of rare cells on said porous medium;

contacting said porous medium with a nutrient-rich elution-fluid comprising a viscosity increasing polymer and at least one complement complex product; and

selectively eluting said first population of rare cells from said porous medium.

51. The method of claim 50, wherein said complement complex product is selected from the group consisting of C3a, C4a, and C5a.

52. A method of harvesting rare cells from a blood product comprising:

contacting a blood product containing rare cells comprising leukocytes with a porous medium comprising a fibrous web;

selectively retaining said rare cells on said porous medium;

backflushing said porous medium with a nutrient-rich elution fluid including a viscosity increasing polymer comprising dextran; and

selectively eluting said rare cells from said porous medium.