Method of preparing an undifferentiated cell

• US 20030166272A1
• Filed: 05/16/2002
• Published: 09/04/2003
• Est. Priority Date: 02/02/1995
• Status: Active Grant

• Current Assignee: Ilham Saleh Abuljadayel
• Original Assignee: Ilham Saleh Abuljadayel
• Inventors: Abuljadayel, Ilham Saleh
• Granted Patent: US 7,410,773 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C12N 2500/14 Calcium; Ca chelators; Calcitonin
• CPC Class Codes: C12N 2500/24 Iron; Fe chelators; Transferrin
• CPC Class Codes: C12N 2501/06 Anti-neoplasic drugs, anti-retroviral drugs, e.g. azacytidine, cyclophosphamide
• CPC Class Codes: C12N 2501/14 Erythropoietin [EPO]
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/50 Cell markers; Cell surface determinants
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

A method of increasing the relative number of cells expressing one or more stem cell markers in a cell population including committed cells is described. The method comprises: i. contacting the cell population with an agent that operably engages said committed cells; and ii. incubating committed cells that are engaged by said agent such that the relative number of cells expressing one or more stem cell markers increases as a result of said engaging.

Claims

1. A method of increasing the relative number of cells expressing one or more stem cell markers in a cell population including committed cells, which method comprises:

i. contacting the cell population with an agent that operably engages said committed cells; and

ii. incubating committed cells that are engaged by said agent such that the relative number of cells expressing one or more stem cell markers increases as a result of said engaging.

2. The method according to claim 1 wherein the committed cells are non-cancer cells.

3. The method according to claim 1 wherein the committed cells are differentiated cells.

4. The method according to claim 3 wherein the committed cells are selected from CFC-T cells, CFC-B cells, CFC-Eosin cells, CFC-Bas cells, CFC-GM cells, CFC-MEG cells, BFC-E cells, CFC-E cells, T cells and B cells.

5. The method according to claim 1 wherein the cells expressing one or more stem cell markers are pluripotent stem cells.

6. The method according to claim 1 wherein the cells expressing one or more stem cell markers are stem cells selected from the group consisting of haemopoietic stem cells, neuronal stem cells, epithelial stem cell, mesenchymal stem cells and embryonic stem cells.

7. The method according to claim 1 wherein the cells expressing one or more stem cell markers are MHC class I⁺ and/or MHC class II⁺ cells.

8. The method according to claim 1 wherein the agent engages a receptor that mediates capture, recognition or presentation of an antigen at the surface of the committed cells.

9. The method according to claim 8 wherein the receptor is an MHC class I antigen or an MHC class II antigen.

10. A method according to claim 9 wherein said class I antigen is an HLA-A receptor, an HLA-B receptor, an HLA-C receptor, an HLA-E receptor, an HLA-F receptor or an HLA-G receptor and said class II antigen is an HLA-DM receptor, an HLA-DP receptor, an HLA-DQ receptor or an HLA-DR receptor.

11. The method according to claim 10 wherein the receptor is an HLA-DR receptor.

12. The method according to claim 8 wherein the receptor comprises a β

-chain having homologous regions.

13. The method according to claim 12 wherein the receptor comprises at least the homologous regions of the β

-chain of HLA-DR.

14. The method according to claim 8 wherein the agent is an antibody to the receptor.

15. A method according to claim 14 wherein the agent is a monoclonal antibody to the receptor.

16. A method according to claim 15 wherein the antibody is selected from the group consisting of monoclonal antibody CR3/43 and monoclonal antibody TAL 1B5.

17. A method according to claim 1 wherein the agent modulates MHC gene expression.

18. A method according to claim 17 wherein the agent modulates MHC Class I⁺ and/or MHC Class II⁺ expression.

19. A method according to claim 1 wherein the agent is used in conjunction with a biological response modifier.

20. A method according to claim 19 wherein the biological response modifier is selected from the group consisting of an alkylating agent, an immunomodulator, a growth factor, a cytokine, a cell surface receptor, a hormone, a nucleic acid, a nucleotide sequence, an antigen and a peptide.

21. A method according to claim 20 wherein the alkylating agent is or comprises cyclophosphoamide.

22. A method according to claim 1 wherein the method is an in vitro method.

23. A method according to claim 1 further comprising recommitting the cells expressing one or more stem cell markers into more differentiated cells.

24. A method according to claim 23 wherein the more differentiated cells are of the same lineage as the committed cells.

25. A method according to claim 23 wherein the more differentiated cells are of a different lineage than the committed cells.

26. Cells expressing one or more stem cell markers produced according to the method of claim 1.

27. A recommitted cell produced according to the method of claim 23.

28. The method according to claim 1 further comprising enriching said cells expressing one or more stem cell markers or recovering said cells expressing one or more stem cell markers from the cell population by using a cell surface marker.

29. The method according to claim 28 wherein said cells expressing one or more stem cell markers are recovered from the cell population after about 2 hours.

50. The method of claim 1 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

63. The method of claim 1 wherein said stem cell marker is selected from the group consisting of CD34, CD19, CD3, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, Nestein, SH2, SH3, CD29, CD44, CD71, CD90, CD106, CD120a, CD124, OCT-4, HLA-DR⁻

, CD38⁻

, CD45⁻

, CD10, CD45low, Rag1, Rag2, high molecular weight glycoprotein TRA-1-60, high molecular weight glycoprotein TRA-1-81 and alkaline phosphatase.

30. A method of producing an altered cell population comprising cells expressing one or more stem cell markers capable of being recommitted into more differentiated cells, which method comprises:

(i) contacting an initial cell population comprising committed cells with an agent that operably engages said committed cells; and

(ii) culturing committed cells within the cell population that are engaged by said agent to become cells expressing one or more stem cell markers as a result of said engaging, thereby resulting in an altered cell population comprising said cells expressing one or more stem cell markers; and

optionally

(iii) enriching said cells expressing one or more stem cell markers or recovering said cells expressing one or more stem cell markers from the altered cell population.

51. The method of claim 30 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

31. A method of producing cells expressing one or more stem cell markers having one or more stem cell markers from committed cells, said cells expressing one or more stem cell markers being capable of being recommitted into more differentiated cells, which method comprises:

(i) contacting an initial cell population comprising the committed cells with an agent that operably engages the surface of said differentiated cells; and

(ii) culturing committed cells that are engaged by said agent to become cells expressing one or more stem cell markers as a result of said engaging, thereby producing cells expressing one or more stem cell markers from committed cells and resulting in an altered cell population comprising sa id cells expressing one or more stem cell markers; and

optionally

(iii) enriching said cells expressing one or more stem cell markers or recovering said cells expressing one or more stem cell markers from the altered cell population.

32. The method according to claim 31 wherein step (iii) comprises enriching said cells expressing one or more stem cell markers or recovering said cells expressing one or more stem cell markers from the altered cell population by using a cell surface marker.

33. The method according to claim 31 wherein said cells expressing one or more stem cell markers are recovered from the altered cell population after about 2 hours.

52. The method of claim 31 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

34. A method of inducing in a cell population committed cells of a hemopoietic lineage to become cells of another lineage which method comprises:

(i) contacting the cell population with an agent that operably engages said committed cells; and

(ii) culturing committed cells that are engaged by said agent to become cells of another lineage as a result of said engaging.

35. A method according to claim 34 wherein said committed cells are of a B cell lineage and become cells of another hemopoietic lineage selected from a T cell lineage and a myeloid lineage.

53. The method of claim 34 wherein said cell of another lineage is optionally isolated and/or purified.

36. A method of producing stem cells from B cells, which method comprises:

(i) contacting an initial cell population comprising B cells with an agent that engages the β

chain of an MHC class II antigen present on the surface of said B cells; and

(ii) culturing B cells that are engaged by said agent to become stem cells as a result of said engaging, thereby resulting in stem cells from B cells and an altered cell population comprising said stem cells; and

optionally

(iii) enriching said stem cells or recovering said stem cells from the altered cell population.

54. The method of claim 36 wherein said T cell is optionally isolated and/or purified.

37. A method of inducing in a cell population a loss of appearance of one or more cell surface markers of committed cells followed by appearance of one or more cell surface markers of stem cells, which method comprises:

(i) contacting the cell population with an agent that operably engages said committed cells; and

(ii) culturing committed cells that are engaged by said agent to have appearance of one or more cell surface markers of stem cells as a result of said engaging.

38. The method of claim 37 wherein the committed cells are hemopoietic.

39. The method of claim 37 wherein the agent engages a receptor that mediates capture, recognition or presentation of an antigen at the surface of the committed cells.

40. The method according to claim 37 further comprising enriching said stem cells or recovering said stem cells from the cell population by using a cell surface marker.

55. The method of claim 37 wherein said myeloid cell is optionally isolated and/or purified.

41. A method of producing an altered cell population comprising cells having an appearance of one or more cell surface markers of stem cells, which method comprises:

(i) contacting an initial cell population comprising committed cells with an agent that operably engages said committed cells; and

(ii) culturing committed cells within the cell population that are engaged by said agent to become cells having an appearance of one or more cell surface markers of stem cells as a result of said engaging, thereby resulting in an altered cell population comprising said stem cells; and

optionally

(iii) enriching said stem cells or recovering said stem cells from the altered cell population.

56. The method of claim 41 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

42. A method of producing cells having an appearance of one or more cell surface markers of stem cells capable of being recommitted into more differentiated cells, which method comprises:

(i) contacting an initial cell population comprising committed cells with an agent that operably engages the surface of said differentiated cells; and

(ii) culturing committed cells that are engaged by said agent to become cells having an appearance of one or more cell surface markers of stem cells as a result of said engaging, thereby producing cells having an appearance of one or more cell surface markers of stem cells capable of being recommitted into more differentiated cells and resulting in an altered cell population comprising said stem cells; and

optionally

(iii) enriching said stem cells or recovering said stem cells from the altered cell population.

43. A method according to claim 42 further comprising recommitting the cell having the appearance of one or more cell surface markers of a stem cell into more cells having the appearance of one or more cell surface markers of differentiated cells.

44. A cell having the appearance of one or more cell surface markers of a stem cell produced according to the method of claim 42.

45. A cell having the appearance of one or more surface markers of differentiated cells produced according to the method of claim 43.

57. The method of claim 42 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

59. The method of claim 44 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

46. A method for identifying a substance capable of retrodifferentiating committed/differentiated cells to cells expressing one or more stem cell markers, which method comprises contacting a population of cells comprising committed cells with a candidate substance and determining whether there is an increase in the relative numbers of cells expressing one or more stem cell markers in said cell population.

47. The method of claim 46 wherein the committed cell is a hemopoietic cell.

48. The method of claim 47 wherein the hemopoietic cell is selected from CFC-T cells, CFC-B cells, CFC-Eosin cells, CFC-Bas cells, CFC-GM cells, CFC-MEG cells, BFC-E cells, CFC-E cells, T cells and B cells.

58. The method of claim 46 wherein said cells expressing one or more stem cell markers are optionally isolated and/or purified.

49. The method of claim 49 wherein the hemopoietic cell is a B cell.

60. A method of increasing the relative number of cells expressing one or more stem cell markers in a cell population including committed cells, which method comprises:

i. contacting the cell population with an antibody that operably engages said committed cells; and

ii. incubating committed cells that are engaged by said antibody such that the relative number of cells expressing one or more stem cell markers increases as a result of said engaging.

61. A method according to claim 60 wherein the antibody is selected from the group consisting of monoclonal antibody CR3/43 and monoclonal antibody TAL 1B5.

62. The method of claim 60 wherein said cell with stem cell surface marker is optionally isolated and/or purified.

64. The method of claim 60 wherein said stem cell marker is selected from the group consisting of one or more of CD34, CD19, CD3, SSEA-3, SSEA4, TRA-1-60, TRA-1-81, Nestein, SH2, SH3, CD29, CD44, CD71, CD90, CD106, CD120a, CD124, OCT-4, HLA-DR⁻

, CD38⁻

, CD45⁻

, CD10, CD45low, Rag1, Rag2, high molecular weight glycoprotein TRA-1-60, high molecular weight glycoprotein TRA-1-81 and alkaline phosphatase.