PLURIPOTENT STEM CELLS CHARACTERIZED BY EXPRESSION OF GERMLINE SPECIFIC GENES

• US 20100297087A1
• Filed: 04/11/2007
• Published: 11/25/2010
• Est. Priority Date: 04/11/2006
• Status: Abandoned Application

• Current Assignee: Nanodiagnostics Israel, Ltd.
• Original Assignee: Nanodiagnostics Israel, Ltd.
• Inventors: Seligman, Judith
• Application Number:
  US12/295,580
• Publication Number:
  US 20100297087A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0611 Primordial germ cells, e.g. embryonic germ cells [EG]

Abstract

The present invention relates to pluripotent or multipotent somatic stem cells characterized by the expression of the germline specific gene, DAZL. The somatic stem cells expressing the DAZL marker are further characterized by expression of additional markers and absence of expression of certain blood markers. In particular, the present invention discloses therapeutic and diagnostic uses, other than the germ cell potential use, of the DAZL pluripotent or multipotent stem cells isolated from adult or peripheral somatic sources such as peripheral blood, bone marrow or umbilical cord blood.

Claims

1-37. -37. (canceled)

38. An isolated somatic multipotent or pluripotent stem cell characterized by expression of the germline specific gene DAZL and at least one gene selected from the group consisting of SOX-2, OCT-4, c-Kit and Stellar.

39. The isolated stem cell of claim 38, expressing the genes SOX-2, OCT-4, c-Kit and Stellar.

40. The isolated stem cell of claim 38, which does not express at least one of the blood markers selected from the group consisting of CD3, CD38, CD14, CD34 and CD133.

41. The isolated cell of claim 38, wherein the cell is a human cell.

42. The isolated cell of claim 41, wherein the cell is isolated from a cell population selected from the group consisting of peripheral blood, umbilical cord blood, bone marrow, stem cell factor mobilized cells, and colony stimulating factor mobilized cells.

43. The isolated cell of claim 41, wherein the cell is isolated from amniotic fluid.

44. The isolated cell of claim 41, wherein the cell is of fetal origin within a maternal cell population.

45. A method of diagnosis of a genetic disorder or chromosomal abnormality in a fetus which comprises:

selecting at least one stem cell according to claim 38 from a fetus using the DAZL specific marker;

producing a display of the chromosomes of the embryonic stem cell; and

analyzing the displayed chromosomes.

46. The method of claim 45 wherein the at least one stem cell is derived from amniotic fluid.

47. A method of treating a tissue disorder or disease comprising administering to a patient in need thereof a somatic stem cell according to claim 38 and providing conditions for differentiation of said cells into cells characterizing the tissue, thereby treating the individual suffering from the tissue disorder or disease.

48. The method of claim 47 wherein the disorder or disease is selected from the group consisting of hematopoietic disease or disorder, neuronal disease or disorder, endothelial disease or disorder, cartilage or bone disease or disorder, liver disease or disorder and heart disease or disorder.

49. The method of claim 47 which further comprises subjecting the somatic pluripotent or multipotent stem cell to culturing conditions suitable for inducing cell proliferation, thereby obtaining an expanded stem cell population;

and introducing the expanded stem cell population into a tissue of the individual associated with the disorder, thereby treating the individual suffering from the disorder requiring cell or tissue replacement.

50. A method of studying stem cell differentiation comprising monitoring the expansion of a somatic stem cell according to claim 38.

51. A method of isolating pluripotent or multipotent somatic cells from an adult or perinatal source which comprises:

separating a mononuclear cell fraction from a cell population;

introducing a molecular probe targeting the specific marker DAZL and optionally at least one additional marker selecting from the group consisting of Sox-2, Stellar, Oct-4 and c-kit into the mononuclear cell fraction thus obtained;

sorting the cells by means of sorting methodology; and

isolating the stem cells expressing the specific marker DAZL and optionally the at least one of the genes selected from group consisting of Sox-2, Stellar, Oct-4 and c-kit.

52. The method of claim 51 which further comprises removing cells expressing at least one of the blood markers selected from the group consisting of CD3, CD38, CD14, CD34 and CD133.

53. The method of claim 51, wherein the molecular probe is a molecular beacon probe.

54. The method of claim 51, wherein the cell population is selected from the group consisting of:

peripheral blood, umbilical cord blood, bone marrow, stem cell factor mobilized cells, colony stimulating factor mobilized cells, a body fluid, a tissue sample, a tissue culture, an organ sample, an organ culture, a cell line and a cell culture.

55. A kit for isolation, enrichment and detection of multipotent or pluripotent stem cells within a specimen, comprising:

at least one reagent to detect DAZL protein or DAZL RNA;

optionally reagents to detect at least one of the genes or gene products selected from the group consisting of;

Sox-2, Stellar, C-kit, and Oct-4;

instructions for labeling, sorting and enrichment of the cells; and

optionally

means for performing stem cell labeling, sorting and enrichment.

56. The kit of claim 55, further comprising reagents and means for negative selection of cells which express at least one of the blood markers CD3, CD38, CD14, CD34 and CD133.

57. The kit of claim 55 further comprising reagents for genetic analysis of fetal and maternal cells.