Somatic Cells for Use in Cell Therapy

• US 20100105133A1
• Filed: 08/03/2006
• Published: 04/29/2010
• Est. Priority Date: 08/03/2005
• Status: Active Grant

• Current Assignee: Core Dynamics Limited
• Original Assignee: Core Dynamics Limited
• Inventors: Kanias, Tamir, Natan, Yehudit
• Granted Patent: US 8,198,085 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0221 Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/545 Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells

Abstract

The present invention generally concerns cell therapy and products for use in such therapy. Particularly, the invention provides a preserved cell preparation essentially free of one or more members of a group of cryoprotecting agents consisting of polyalcohols, DMSO and cryoprotecting proteins, the preserved cell preparation comprising somatic cells and at least one polyphenol, wherein upon reconstitution of cells in the cell preparation, at least a portion of said stem cells are viable, said portion being sufficient for use of the cell preparation in stem cell therapy. The invention also provides cells reconstituted from preserved somatic cells, and the use of the reconstituted cells in cell therapy. A preferred cell preparation in accordance with the invention comprises stem cells, preferably human stem cells.

Claims

1. A preserved cell preparation essentially free of one or more members of a group of cryoprotecting agents consisting of polyalcohols, DMSO and cryoprotecting proteins, the preserved cell preparation comprising somatic cells and at least one polyphenol, wherein upon reconstitution of cells in the cell preparation, at least a portion of said stem cells are viable, said portion being sufficient for use of the cell preparation in stem cell therapy.

2. The preserved cell preparation of claim 1, wherein preservation comprises hypothermic conditions, freezing or freeze-drying and combinations of same.

3. The preserved cell preparation of claim 2, wherein after thawing of the frozen cell preparation at least 90% of the somatic cells are viable as compared to the total number of cells in the cell preparation prior to freezing.

4. The preserved cell preparation of claim 2, wherein after re-hydrating the freeze-dried cell preparation at least 60% of the somatic cells are viable as compared to the total number of cells in the cell preparation prior to freeze-drying.

5. The preserved cell preparation of any one of claims 1 to 4, wherein said polyphenol is a synthetic or naturally derived catechin selected from epigallocatechin gallate (EGCG), epicatechin gallate (ECG) epigallocatechin (EGC) epicatechin (EC), DL-Catechin (DL-C) gallocatechin gallate (GCG), and mixtures of same.

6. The preserved cell preparation of claim 5, wherein said polyphenol is EGCG.

7. The preserved cell preparation of claim 6, comprising upon reconstitution or before freezing between 50 μ

M to 4 mM EGCG.

8. The preserved cell preparation of any one of claims 1 to 7, comprising a cryoprotecting agent.

9. The preserved cell preparation of claim 8, wherein said cryoprotecting agent is trehalose.

10. The preserved cell preparation of any one of claims 1 to 9 comprising EGCG and trehalose.

11. The preserved cell preparation of claim 10 comprising upon reconstitution or before freezing EGCG at a concentration between about 0.5 mg/ml to about 1 mg/ml.

12. The preserved cell preparation of claim 11 comprising upon reconstitution or before freezing EGCG at a concentration of about 0.945 mg/ml.

13. The preserved cell preparation of any one of claims 1 to 12, obtainable by cooling the cells at a cooling rate of 5.1°

C./min.

14. The preserved cell preparation of claim 12, obtainable by freezing said cells followed by freeze drying of the frozen cells to obtain a dry preserved cell preparation.

15. The preserved cell preparation of any one of claims 1 to 12, comprising stem cells.

16. The preserved cell preparation of claim 14, wherein said stem cells are selected from hematopoietic stem cells, embryonic stem cells, embryonic germ cells and mesenchymal stem cells.

17. The preserved cell preparation of claim 15, wherein said cells are human stem cells.

21. The use of any one of claims 18 to 20, wherein said preserved cell preparation is as defined in any one of claims 2 to 17.

41. A somatic cell transplantation kit comprising a preserved cell preparation according to any one of claims 1 to 16 and instructions for reconstituting the preserved cell preparation.

18. Use of a preserved cell preparation for the production of a therapeutic composition for cell therapy, the preserved cell preparation being essentially free of one or more members of a group of cryoprotecting agents consisting of polyalcohols, DMSO and cryoprotecting proteins, the preserved cell preparation comprising somatic cells, wherein upon reconstitution of cells in the cell preparation, at least a portion of said cells are viable, said portion being sufficient for use of the therapeutic composition in cell therapy.

19. The use of claim 18, wherein said somatic cells comprise stem cells.

20. The use of claim 18 or 19, in combination with at least one physiologically acceptable additive suitable for transplantation of said cell preparation in a mammalian subject.

22. A reconstituted cell preparation being essentially free of one or more members of a group of cryoprotecting agents comprising polyalcohol, DMSO and cryoprotecting proteins, the reconstituted cell preparation comprising post-preservation somatic cells and at least one polyphenol, wherein at least a portion of said post-preservation somatic cells are viable, said portion being sufficient for use of the reconstituted cell preparation in cell therapy.

23. The reconstituted cell preparation of claim 22, wherein said somatic cells comprise stem cells.

24. The reconstituted cell preparation of claim 23, wherein reconstitution of said cells comprises warming, thawing and/or re-hydrating said cells.

25. The reconstituted cell preparation of claim 24, wherein at least 20% of the recovered cells are viable.

26. The reconstituted cell preparation of claim 25, wherein at least 60% of the recovered cells are viable.

27. The reconstituted cell preparation of claim 23, wherein at least 90% of the cells are viable as compared to the total number of cells in the cell preparation prior to freezing.

28. The reconstituted cell preparation of claim 24, wherein at least 60% of the cells are viable as compared to the total number of cells in the cell preparation prior to freeze-drying.

29. The reconstituted cell preparation of any one of claims 22 to 28, wherein said polyphenol is a synthetic or naturally derived catechin selected from epigallocatechin gallate (EGCG), epicatechin gallate (ECG) epigallocatechin (EGC) epicatechin (EC), DL-Catechin (DL-C) gallocatechin gallate (GCG), and mixtures of same.

30. The reconstituted cell preparation of claim 29, wherein said polyphenol is EGCG.

31. The reconstituted cell preparation of claim 30, wherein said EGCG is present in an amount of between 50 μ

M to 4 mM EGCG.

32. The reconstituted cell preparation of claim 31, wherein said EGCG is present at a concentration of between about 0.5 mg/ml to about 1.0 mg/ml.

33. The reconstituted cell preparation of claim 32, wherein said EGCG is present at a concentration of about 0.945 mg/ml.

34. The reconstituted cell preparation of any one of claims 22 to 33, comprising stem cells.

35. The reconstituted cell preparation of claim 34, wherein the stem cells are selected from hematopoietic stem cells, embryonic stem cells, mesenchymal stem cells, and embryonic germ cells.

36. The reconstituted cell preparation of any one of claims 22 to 35, wherein said cells are human cells.

40. The method of claim 35 wherein the cell preparation comprises human cells.

37. A method of treating a subject in need of somatic cell transplantation, the method comprises administering to said subject a cell preparation reconstituted from preservation conditions, the cell preparation being essentially free of one or more members of a group of cryoprotecting agents consisting of polyalcohols, DMSO and cryoprotecting proteins, the cell preparation comprising a effective amount of post-preservation and viable somatic cells the amount of the viable cells being sufficient for establishing said somatic cells in the subjects body.

38. The method of claim 37, wherein the cell preparation comprises stem cells.

39. The method of claim 38, wherein said stem cells are selected from hematopoietic stem cells, embryonic stem cells, mesenchymal stem cells or embryonic germ cells.