Stem Cells Suitable for Transplantation, their Preparation and Pharmaceutical Compositions Comprising Them

• US 20080025957A1
• Filed: 11/08/2004
• Published: 01/31/2008
• Est. Priority Date: 11/13/2003
• Status: Abandoned Application

• Current Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Original Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Inventors: Kahn, Joy, Lapidot, Tsvee
• Application Number:
  US10/578,291
• Publication Number:
  US 20080025957A1
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/7158 for chemokines
• CPC Class Codes: C12N 2501/21 Chemokines, e.g. MIP-1, MIP-2, RANTES, MCP, PF-4
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The present invention relates to stem cells suitable for transplantation and to methods for their preparation.

Claims

1. A method for preparing a population of cells comprising stem cells with a high amount of immature primitive progenitors, the method comprising collecting stem cells and introducing into the cells a DNA fragment comprising the sequence of CXCR4.

2. A method according to claim 1, wherein the population of cells exhibits improved CXCR4 signaling in response to low concentration of SDF-1.

3. A method according to claim 1, wherein the population of cells exhibits improved CXCR4 signaling in response to high concentration of SDF-1.

4. The method according to claim 1, wherein the stem cells are hematopoietic stem cells.

5. The method according to claim 4, wherein the hematopoietic stem cells are CD34⁺ enriched.

6. The method according to claim 1, wherein the immature primitive progenitors are of the CD34⁺/CD38 ⁻

/low lineage.

7. The method according to claim 1, wherein collecting the stem cells is effected after inducing stem cell mobilization procedure;

and/or comprises a surgical procedure.

8. The method according to claim 1, further comprising isolating by FACS stem cells having CXCR4 levels above a predetermined threshold.

9. The method according to claim 1, wherein the stem cells are capable of differentiating towards the myeloid and erythroid lineages.

10. The method according to claim 6, wherein the amount of immature primitive progenitors of the CD34⁺/CD38 ⁻

/low lineage are about 1-5% of the population.

11. The method according to claim 6, wherein the immature primitive progenitors of the CD34⁺/CD38 ⁻

/low lineage are in an amount equal to or higher than about 3% of the population.

12. The method according to claim 2, wherein the low concentration of SDF-1 is equal to or lower than about 50 ng/ml.

13. The method according to claims 2 or 12, wherein improved signaling is manifested as the enhancement of cell migration mediated by low concentrations of SDF-1.

14. The method according to claims 2 or 12, wherein improved signaling is manifested as the enhancement of cell proliferation mediated by low concentrations of SDF-1.

15. The method according to claim 3, wherein the high concentration of SDF-1 is equal to or higher than about 1 microgram/ml.

16. The method according to claims 3 or 15, wherein improved signaling signaling is manifested by a reduction in desensitization by SDF-1.

17. A population comprising stem cells expressing a high amount of immature primitive progenitors, exhibiting improved CXCR4 signaling capability in response to low and/or high concentrations of SDF-1, prepared by introducing into the stem cells a DNA fragment comprising the sequence of CXCR4.

18. The population of stem cells according to claim 17, wherein the stem cells are hematopoietic stem cells.

19. The population of cells according to claim 17 or 18, being capable of differentiating towards the myeloid and erythroid lineages.

20. The population of cells according to claim 19, wherein the hematopoietic stem cells are CD34⁺ hematopoietic stem cells.

21. The population of cells according to claim 17, wherein the immature primitive progenitors are of the CD34⁺/CD38 ⁻

/low lineage.

22. The population of cells according to claim 21, wherein high amount of CD34⁺/CD38 ⁻

/low is about 1-5% of the population.

23. The population of cells according to claim 21, wherein the high amount of CD34⁺/CD³⁸⁻

/low is about and above 3% of the population.

24. The population of cells according to claim 17, wherein the low concentration of SDF-1 is about and below 50 ng/ml.

25. The population of cells according to claim 17, wherein the high concentration of SDF-1 is equal to or higher than about 1 microgram/ml.

35. A method of treating a disorder in a subject requiring cell or tissue replacement, the method comprising providing to a subject in need thereof a therapeutically effective amount of a population of cells according to anyone of claims 17 to 25.

26. A method for increasing homing of stem cells to a target tissue in a subject in need thereof, comprising administering to said subject a population of cells comprising stem cells expressing a high amount of immature primitive progenitors, exhibiting improved CXCR4 signaling capability in response to low and/or high concentrations of SDF-1, prepared by introducing into the stem cells a DNA fragment comprising the sequence of CXCR4.

28. The method according to claims 26 or 27, wherein said target tissue is selected from the group consisting of bone marrow, blood vessel, heart, lung, liver, pancreas, kidney, nervous system, skin, bone and skeletal muscle.

29. The method according to claims 26 or 27, to facilitate transplantation.

30. The method according to claim 29, wherein transplantation follows chemotherapy protocols.

31. The use method according to claim 29, wherein transplantation is autologous.

32. The method according to claim 31, wherein the transplantation involves mobilization of autologous cells.

33. The method according to claim 29, wherein transplantation is heterologous.

34. The method according to claim 29, wherein the transplantation is carried with mobilized stem cells.

27. A method for increasing repopulation of a target tissue in a subject in need thereof, comprising administering to said subject a population of cells comprising stem cells expressing a high amount of immature primitive progenitors, exhibiting improved CXCR4 signaling capability in response to low and/or high concentrations of SDF-1, prepared by introducing into the stem cells a DNA fragment comprising the sequence of-CXCR4.

36. A method of preparing a population of cells comprising stem cells exhibiting CXCR4 with intact 6H8 epitope, the method comprising collecting stem cells and introducing to the cells a DNA fragment comprising the sequence of CXCR4.

37. A population of cells comprising stem cells comprising intact CXCR4 6H8 epitope prepared by introducing a DNA fragment comprising the sequence of CXCR4.

38. A method for for transplantation in a subject in need thereof, comprising contacting said subject with the population of cells of claim 37.

39. A method of treating a disorder requiring cell or tissue replacement, the method comprising providing to a subject in need thereof a therapeutically effective amount of a population of cells according to claim 37.

40. A pharmaceutical composition comprising a population of stem cells comprising stem cells exhibiting intact CXCR4 6H8 epitope prepared by introducing to the cells a DNA fragment comprising the sequence of CXCR4.