Nuclear reprogramming factor and induced pluripotent stem cells

• US 20090227032A1
• Filed: 11/06/2008
• Published: 09/10/2009
• Est. Priority Date: 12/13/2005
• Status: Abandoned Application

• Current Assignee: Kyoto University
• Original Assignee: Kyoto University
• Inventors: Yamanaka, Shinya, Takahashi, Kazutoshi, Nakagawa, Masato
• Application Number:
  US12/289,873
• Publication Number:
  US 20090227032A1
• Time in Patent Office: Days
• US Class Current: 435/456
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 2501/604 Klf-4
• CPC Class Codes: C12N 2501/606 c-Myc
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2740/15043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The present invention relates to a nuclear reprogramming factor having an action of reprogramming a differentiated somatic cell to derive an induced pluripotent stem (iPS) cell. The present invention also relates to the aforementioned iPS cells, methods of generating and maintaining iPS cells, and methods of using iPS cells, including screening and testing methods as well as methods of stem cell therapy. The present invention also relates to somatic cells derived by inducing differentiation of the aforementioned iPS cells.

Claims

1. A pluripotent stem cell induced by reprogramming a somatic cell, wherein the reprogramming is performed in the absence of eggs, embryos, or embryonic stem (ES) cells and comprises contacting the somatic cell with a nuclear reprogramming factor comprising one or more gene products of a Sall1 gene and/or a Sall4 gene.

2. The pluripotent stem cell of claim 1, wherein the somatic cell is a human cell.

3. The pluripotent stem cell of claim 1, wherein the somatic cell is a mouse cell.

4. The pluripotent stem cell of claim 1, wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell.

5. The pluripotent stem cell of claim 1, wherein the nuclear reprogramming factor comprises one or more gene products of a human or mouse Sall4 gene.

6. The pluripotent stem cell of claim 1, wherein the nuclear reprogramming factor comprises one or more gene products of each of:

an Oct family gene, a Klf family gene, and a Sox family gene.

7. The pluripotent stem cell of claim 6, wherein the nuclear reprogramming factor comprises one or more gene products of a Myc family gene.

8. The pluripotent stem cell of claim 6, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and Sox2, and at least one of Sall1 and Sall4.

9. The pluripotent stem cell of claim 7, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and c-Myc, and at least one of Sall1 and Sall4.

10. The pluripotent stem cell of claim 1, wherein the nuclear reprogramming factor comprises one or more gene products of each of:

an Oct family gene, a Sox family gene, and a Nanog family gene.

11. The pluripotent stem cell of claim 10, wherein the nuclear reprogramming factor includes one or more gene products of a Lin28 family gene.

12. The pluripotent stem cell of claim 10, wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, and Nanog, and at least one of Sall1 and Sall4.

13. The pluripotent stem cell of claim 11, wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, Nanog, and Lin28, and at least one of Sall1 and Sall4.

67. A somatic cell derived by inducing differentiation of the pluripotent stem cell according to claim 1.

14. A pluripotent stem cell induced by reprogramming a somatic cell, wherein the reprogramming is performed in the absence of eggs, embryos, or embryonic stem (ES) cells and comprises contacting the somatic cell with a nuclear reprogramming factor comprising one or more gene products of:

an L-Myc gene, an N-Myc gene, and/or a c-Myc gene.

15. The pluripotent stem cell of claim 14, wherein the somatic cell is a human cell.

16. The pluripotent stem cell of claim 14, wherein the somatic cell is a mouse cell.

17. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell.

18. The pluripotent stem cell of claim 14, wherein the L-Myc gene, the N-Myc gene, and/or the c-Myc gene is a chimera or mutant gene.

19. The pluripotent stem cell of claim 18, wherein the nuclear reprogramming factor comprises Ms-cL-Myc or its human homolog.

20. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor comprises amino acids 1-144 of c-Myc or its human homolog.

21. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor comprises amino acids 105-277 of L-Myc or its human homolog.

22. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor comprises amino acids 347-439 of c-Myc, amino acids 278-368 of L-Myc, or their human homolog.

23. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor comprises one or more gene products of:

an Oct family gene, and a Klf family gene.

24. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor comprises one or more gene products of a Sox family gene.

25. The pluripotent stem cell of claim 14, wherein the nuclear reprogramming factor comprises one or more gene products of a Lin28 family gene.

26. The pluripotent stem cell of claim 23, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and L-Myc.

27. The pluripotent stem cell of claim 24, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and L-Myc.

28. The pluripotent stem cell of claim 25, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, L-Myc, and Lin28.

68. A somatic cell derived by inducing differentiation of the pluripotent stem cell according to claim 14.

29. A pluripotent stem cell induced by reprogramming a somatic cell, wherein the reprogramming is performed in the absence of eggs, embryos, or embryonic stem (ES) cells and comprises contacting the somatic cell with a nuclear reprogramming factor comprising one or more gene products of a Lin28 gene.

30. The pluripotent stem cell of claim 29, wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell.

69. A somatic cell derived by inducing differentiation of the pluripotent stem cell according to claim 29.

31. A method for preparing an induced pluripotent stem cell by nuclear reprogramming of a somatic cell, which comprises contacting a nuclear reprogramming factor comprising one or more gene products of a Sall1 gene and/or a Sall4 gene with the somatic cell to obtain an induced pluipotent stem cell.

32. The method of claim 31, wherein the somatic cell is a human cell.

33. The method of claim 31, wherein the somatic cell is a mouse cell.

34. The method of claim 31, wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell.

35. The method of claim 31, wherein the nuclear reprogramming factor comprises one or more gene products of a human or mouse Sall4 gene.

36. The method of claim 31, wherein the nuclear reprogramming factor comprises one or more gene products of each of:

an Oct family gene, a Klf family gene, and a Sox family gene.

37. The method of claim 36, wherein the nuclear reprogramming factor comprises one or more gene products of a Myc family gene.

38. The method of claim 36, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and Sox2, and at least one of Sall1 and Sall4.

39. The method of claim 37, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and c-Myc, and at least one of Sall1 and Sall4.

40. The method of claim 31, wherein the nuclear reprogramming factor comprises one or more gene products of each of:

an Oct family gene, a Sox family gene, and a Nanog family gene.

41. The method of claim 40, the efficiency of the iPS cell preparation is increased by the addition to the nuclear reprogramming factor of one or more gene products of a Lin28 gene.

42. The method of claim 40, wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, and Nanog, and at least one of Sall1 and Sall4.

43. The method of claim 41, wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, Nanog, and Lin28, and at least one of Sall1 and Sall4.

61. An induced pluripotent stem cell obtained by the method of claim 31.

64. A somatic cell derived by inducing differentiation of the pluripotent stem cell according to claim 61.

44. A method for preparing an induced pluripotent stem cell by nuclear reprogramming of a somatic cell, which comprises contacting a nuclear reprogramming factor comprising one or more gene products of:

an L-Myc gene, an N-Myc gene, and/or a c-Myc gene with the somatic cell to obtain an induced pluipotent stem cell.

45. The method of claim 44, wherein the somatic cell is a human cell.

46. The method of claim 44, wherein the somatic cell is a mouse cell.

47. The method of claim 44, wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell.

48. The method of claim 44, wherein the L-Myc gene, the N-Myc gene, and/or the c-Myc gene is a chimera or mutant gene.

49. The method of claim 48, wherein the nuclear reprogramming factor comprises Ms-cL-Myc or its human homolog.

50. The method of claim 44, wherein the nuclear reprogramming factor comprises amino acids 1-144 of c-Myc or its human homolog.

51. The method of claim 44, wherein the nuclear reprogramming factor comprises amino acids 105-277 of L-Myc or its human homolog.

52. The method of claim 44, wherein the nuclear reprogramming factor comprises amino acids 347-439 of c-Myc, amino acids 278-368 of L-Myc, or their human homolog.

53. The method of claim 44, wherein the nuclear reprogramming factor comprises one or more gene products of:

an Oct family gene, and a Klf family gene.

54. The method of claim 44, wherein the nuclear reprogramming factor comprises one or more gene products of a Sox family gene.

55. The method of claim 44, wherein the nuclear reprogramming factor comprises one or more gene products of a Lin28 family gene.

56. The method of claim 53, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and L-Myc.

57. The method of claim 54, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and L-Myc.

58. The method of claim 55, wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, L-Myc, and Lin28.

62. An induced pluripotent stem cell obtained by the method of claim 44.

65. A somatic cell derived by inducing differentiation of the pluripotent stem cell according to claim 62.

59. A method for preparing an induced pluripotent stem cell by nuclear reprogramming of a somatic cell, which comprises contacting a nuclear reprogramming factor comprising one or more gene products of a Lin28 gene with the somatic cell to obtain an induced pluipotent stem cell.

60. The method of claim 59, wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell.

63. An induced pluripotent stem cell obtained by the method of claim 59.

66. A somatic cell derived by inducing differentiation of the pluripotent stem cell according to claim 63.