Wharton's jelly mesenchymal stem cells and uses thereof

Patent metadata

Field	Value
Publication number	US-9315776-B2
Application number	US-201213667370-A
Country	US
Kind code	B2
Filing date	Nov 2, 2012
Priority date	Nov 9, 2011
Publication date	Apr 19, 2016
Grant date	Apr 19, 2016

Abstract

In one aspect, the invention is directed to methods of expanding hematopoietic stem cells (HSCs) comprising culturing the HSCs with Wharton's Jelly mesenchymal stem cells (WJSCs), a cell culture medium that has been conditioned with WJSCs, or a combination thereof, thereby producing a HSC culture; and maintaining the HSC culture under conditions in which the HSCs expand in the culture, thereby expanding the HSCs. In another aspect, the invention is directed to a method of transplanting the expanded HSCs in an individual in need thereof. In yet another aspect, the invention is directed to compositions comprising HSCs and Wharton's Jelly mesenchymal stem cells (WJSCs). The composition can further comprise a cell culture medium that has been conditioned with WJSCs.

First claim

What is claimed is:

1. A method of expanding hematopoietic stem cells (HSCs), comprising:
(a) culturing an isolated population of HSCs with a composition comprising a cell culture medium that has been conditioned with Wharton's Jelly mesenchymal stem cells (WJSCs), wherein the cell culture medium does not include Wharton's Jelly stem cells (WJSCs), thereby producing a HSC culture; and
(b) maintaining the HSC culture under conditions in which the HSCs expand in the culture, thereby expanding the HSCs.

2. The method of claim 1 wherein the cell culture medium is a basal medium.

3. The method of claim 1 wherein the cell culture medium is an enriched medium.

4. The method of claim 3 wherein the enriched medium comprises one or more cytokines that support growth of HSCs.

5. The method of claim 1 wherein the cell culture medium conditioned with WJSCs is obtained by culturing WJSCs in the medium for about 12 hours to about 48 hours.

6. The method of claim 1 wherein the conditions in which the HSCs expand in culture comprise maintaining the HSC culture at 37° C. in 5% CO 2 .

7. The method of claim 1 wherein the HSCs are cultured for about 1 day, 5 days, 10 days or about 14 days.

8. The method of claim 1 wherein the HSCs are obtained from bone marrow, cord blood, peripheral blood, or a combination thereof.

9. The method of claim 1 wherein the HSCs and the WJSCs are obtained from different individuals.

10. The method of claim 1 wherein the HSCs and WJSCs are obtained from the same individual.

11. The method of claim 1 wherein the HSCs are human HSCs.

12. The method of claim 1 wherein the WJSCs are human WJSCs.

13. The method of claim 1 wherein the HSCs are expanded to about 10-fold or to about 25-fold compared to a suitable control.

14. The method of claim 1 further comprising separating the expanded HSCs from the culture medium that has been conditioned with the WJSCs, thereby obtaining isolated HSCs.

15. The method of claim 14 further comprising storing the isolated HSCs.

16. The method of claim 1 further comprising transplanting an effective amount of the expanded hematopoietic stem cells into an individual in need thereof.

17. The method of claim 16 wherein the individual has a malignant hematopoietic disease.

18. The method of claim 16 wherein the individual has a non-malignant hematopoietic disease.

19. The method of claim 1 , wherein the HSCs are fetal liver HSCs.

20. A method of transplanting hematopoietic stem cells (HSCs) in an individual in need thereof, comprising:
expanding an isolated population of HSCs ex vivo comprising culturing the HSCs with a composition comprising a cell culture medium that has been conditioned with Wharton's Jelly mesenchymal stem cells (WJSCs), wherein the cell culture medium does not include Wharton's Jelly stem cells (WJSCs), thereby producing a HSC culture;
maintaining the HSC culture under conditions in which the HSCs expand in the culture, thereby expanding the HSCs and producing expanded HSCs; and
introducing the expanded HSCs into the individual in need thereof, thereby transplanting HSCs in the individual.

21. The method of claim 20 wherein the expanded HSCs are the individual's own HSCs.

22. The method of claim 20 wherein the individual has a malignant hematopoietic disease.

23. The method of claim 20 wherein the individual has a non-malignant hematopoietic disease.

24. The method of claim 20 wherein the cell culture medium is a basal medium.

25. The method of claim 20 wherein the cell culture medium is an enriched medium.

26. The method of claim 25 wherein the enriched medium comprises one or more cytokines that support growth of HSCs.

27. The method of claim 20 wherein the cell culture medium conditioned with WJSCs is obtained by culturing WJSCs in the medium for about 12 hours to about 48 hours.

28. The method of claim 20 wherein the conditions in which the HSCs expand in culture comprise maintaining the HSC culture at 37° C. in 5% CO 2 .

29. The method of claim 20 wherein the HSCs are cultured for about 1 day, 5 days, 10 days or about 14 days.

30. The method of claim 20 wherein the HSCs are obtained from bone marrow, cord blood, peripheral blood, or a combination thereof.

31. The method of claim 20 wherein the HSCs and the WJSCs are obtained from different individuals.

32. The method of claim 20 wherein the HSCs and WJSCs are obtained from the same individual.

33. The method of claim 20 wherein the HSCs are human HSCs.

34. The method of claim 20 wherein the WJSCs are human WJSCs.

35. The method of claim 20 wherein the HSCs are expanded to about 10-fold or to about 25-fold compared to a suitable control.

36. The method of claim 20 further comprising separating the expanded HSCs from the culture medium that has been conditioned with the WJSCs, thereby obtaining isolated HSCs.

37. The method of claim 20 wherein the individual is a human.

38. The method of claim 20 , wherein the HSCs are fetal liver HSCs.

39. A method of transplanting hematopoietic stem cells (HSCs) in an individual in need thereof, comprising:
providing an expanded population of HSCs, wherein the HSCs were expanded by culturing an isolated population of HSCs with a composition comprising a cell culture medium that has been conditioned with Wharton's Jelly mesenchymal stem cells (WJSCs), and wherein the cell culture medium does not include Wharton's Jelly stem cells (WJSCs); and
introducing the expanded HSCs into the individual in need thereof, thereby transplanting HSCs in the individual.

Assignees

• Univ Singapore

Inventors

• Fong Chui Yee
• Bongso Tuan Ariffeen
• Biswas Arijit

Classifications

• C12N2502/1388

  Mesenchymal stem cells from other natural sources · CPC title

• C12N5/0668

  Mesenchymal stem cells from other natural sources · CPC title

• A61K35/28

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title

• C12N5/0647 Primary

  Haematopoietic stem cells; Uncommitted or multipotent progenitors · CPC title

• C12N2502/025

  extra-embryonic cells, e.g. amniotic epithelium, placental cells, Wharton's jelly · CPC title