Lineage committed stem cells selected for telomerase promoter activity

• US 20050079608A1
• Filed: 08/04/2004
• Published: 04/14/2005
• Est. Priority Date: 02/05/2002
• Status: Abandoned Application

• Current Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD. (Technion-Israel Institute Of Technology)
• Original Assignee: TECHNION RESEARCH AND DEVELOPMENT FOUNDATION, LTD. (Technion-Israel Institute Of Technology)
• Inventors: Tzukerman, Maty, Skorecki, Karl L.
• Application Number:
  US10/911,682
• Publication Number:
  US 20050079608A1
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2510/02 Cells for production
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The present invention discloses transfected embryonic stem cells capable of providing telomerase positive progenitor cells that are capable of proliferation and maintain telomerase gene promoter activity, and to the isolation and propagation of such populations useful for cell replacement therapy. Improvements in cell replacement therapy utilizing such cells form another embodiment of the invention.

Claims

1. An enriched population of adult stem cells derived from stem cells comprising a plurality of committed progenitor cells stably transfected with a polynucleotide construct comprising a telomerase promoter element operably linked to a sequence encoding a selectable marker, wherein the progenitor cells express telomerase promoter activity.

2. The population of adult stem cells according to claim 1, wherein the polynucleotide construct further encodes a reporter gene.

3. The population of adult stem cells according to claim 2, wherein the reporter gene encodes an optically detectable gene product.

4. The population of adult stem cells according to claim 1, wherein the stem cells are of human origin.

5. The population of adult stem cells according to claim 4, wherein the stem cells are derived from human embryonic stem cells.

6. The population of adult stem cells according to claim 1, wherein the telomerase promoter is of human origin.

7. The population according to claim 1, wherein the selectable marker is an antibiotic resistance gene.

8. The population of adult stem cells according to claim 1 further transfected with a lineage specific promoter operably linked to a gene encoding a selectable marker.

9. The population of adult stem cells according to claim 8, wherein the selectable marker is an antibiotic resistance gene.

10. The population of adult stem cells according to claim 1, wherein the cells are committed to a cell lineage selected from the group consisting of cardiomyocytes, beta islet cells, neuronal cells, hepatic cells, chondrocytes, dermal and epidermal cells, connective tissue cells, dendritic cells, hematopoietic cells and any other differentiated cell type which is potentially useful in human cell replacement therapy.

11. The population of adult stem cells according to claim 8, wherein the cells are committed to a cell lineage selected from the group consisting of cardiomyocytes, beta islet cells, neuronal cells, hepatic cells, chondrocytes, dermal and epidermal cells, connective tissue cells, dendritic cells, hematopoietic cells and any other differentiated cell type which is potentially useful in human cell replacement therapy.

38. In cell replacement therapy, the improvement which comprises utilizing the enriched population of adult stem cells according to claim 1 for cell replacement.

12. An isolated subpopulation of adult stem cells derived from stem cells comprising a plurality of committed progenitor cells stably transfected with a polynucleotide construct comprising a telomerase promoter element operably linked to a sequence encoding a selectable marker, wherein the progenitor cells express telomerase promoter activity.

13. The subpopulation of adult stem cells according to claim 12, wherein the polynucleotide construct further encodes a reporter gene.

14. The subpopulation of adult stem cells according to claim 13, wherein the reporter gene encodes an optically detectable gene product.

15. The subpopulation of adult stem cells according to claim 12 wherein the stem cells are of human origin.

16. The subpopulation of adult stem cells according to claim 15, wherein the stem cells are derived from human embryonic stem cells.

17. The subpopulation of adult stem cells according to claim 12, wherein the telomerase promoter is of human origin.

18. The subpopulation of adult stem cells according to claim 12, wherein the selectable marker is an antibiotic resistance gene.

19. The subpopulation of adult stem cells according to claim 12, further transfected with a lineage specific promoter operably linked to a gene encoding a selectable marker.

20. The subpopulation of adult stem cells according to claim 19, wherein the selectable marker is an antibiotic resistance gene.

21. The cell subpopulation of adult stem cells according to claim 12, wherein the committed progenitors are specific to a cell lineage selected from the group consisting of cardiomyocytes, beta islet cells, neuronal cells, hepatic cells, chondrocytes, dermal and epidermal cells, connective tissue cells, dendritic cells, hematopoietic cells and any other differentiated cell type which is potentially useful in human cell replacement therapy.

22. The cell subpopulation of adult stem cells according to claim 19, wherein the committed progenitors are specific to a cell lineage selected from the group consisting of cardiomyocytes, beta islet cells, neuronal cells, hepatic cells, chondrocytes, dermal and epidermal cells, connective tissue cells, dendritic cells, hematopoietic cells and any other differentiated cell type which is potentially useful in human cell replacement therapy.

39. In cell replacement therapy, the improvement which comprises utilizing the isolated subpopulation of cells according to claim 12 for cell replacement.

23. A cloned population of adult stem cells derived from stem cells comprising a plurality of committed progenitor cells stably transfected with a polynucleotide construct comprising a telomerase promoter element operably linked to a sequence encoding a selectable marker, wherein the progenitor cells express telomerase promoter activity.

24. The population of adult stem cells according to claim 23, wherein the polynucleotide construct further encodes a reporter gene.

25. The population of adult stem cells according to claim 24, wherein the reporter gene encodes an optically detectable gene product.

26. The population of adult stem cells according to claim 23, wherein the stem cells are of human origin.

27. The population of adult stem cells according to claim 25, wherein the adult stem cells are derived from human embryonic stem cells.

28. The population of adult stem cells according to claim 23, wherein the telomerase promoter is of human origin.

29. The population of adult stem cells according to claim 23, wherein the selectable marker is an antibiotic resistance gene.

30. The population of adult stem cells according to claim 23, wherein the cells are further transfected with a lineage specific promoter operably linked to a gene encoding a selectable marker.

31. The population of adult stem cells according to claim 30, wherein the selectable marker is an antibiotic resistance gene.

32. The population of adult stem cells according to claim 23, wherein the committed progenitors are specific to a cell lineage selected from the group consisting of cardiomyocytes, beta islet cells, neuronal cells, hepatic cells, chondrocytes, dermal and epidermal cells, connective tissue cells, dendritic cells, hematopoietic cells and any other differentiated cell type which is potentially useful in human cell replacement therapy.

33. The population of adult stem cells according to claim 30, wherein the committed progenitors are specific to a cell lineage selected from the group consisting of cardiomyocytes, beta islet cells, neuronal cells, hepatic cells, chondrocytes, dermal and epidermal cells, connective tissue cells, dendritic cells, hematopoietic cells and any other differentiated cell type which is potentially useful in human cell replacement therapy.

40. In cell replacement therapy, the improvement which comprises utilizing the cloned population of cells according to claim 23 for cell replacement.

34. A population of stem cells stably transfected with a polynucleotide construct comprising ectopic telomerase encoding sequences, the cells over-expressing ectopic telomerase, wherein said cells can differentiate while maintaining higher telomerase activity than the telomerase activity of differentiated cells derived from non-transfected stem cells.

35. The population of cells according to claim 34, wherein the stem cells are of human origin.

36. The population of cells according to claim 35, wherein the stem cells are derived from human embryonic stem cells.

37. The population of cells according to claim 34, wherein the telomerase is of human origin.