STEM CELLS CHARACTERIZED BY EXPRESSION OF GERMLINE SPECIFIC GENES

• US 20110158966A1
• Filed: 02/17/2011
• Published: 06/30/2011
• Est. Priority Date: 07/23/2002
• Status: Abandoned Application

• Current Assignee: Nanodiagnostics Israel, Ltd.
• Original Assignee: Nanodiagnostics Israel, Ltd.
• Inventors: Seligman, Judith
• Application Number:
  US13/029,975
• Publication Number:
  US 20110158966A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 16/18 against material from animals or humans
• 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
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism

Abstract

The present invention relates to adult stem cells derived from somatic sources 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 multipotent stem cells isolated from somatic sources such as peripheral blood, bone marrow or umbilical cord blood.

Claims

1. An isolated stem cell derived from a somatic source, 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.

2. The isolated stem cell of claim 1 expressing the genes SOX-2, OCT-4, c-Kit and Stellar.

3. The isolated stem cell of claim 1 which does not express at least one of the blood markers selected from the group consisting of:

CD3, CD38, CD14, CD34 and CD133.

4. The isolated stem cell of claim 1 wherein the cell is multipotent.

5. The isolated cell of claim 1, wherein the cell is a human cell.

6. The isolated cell of claim 5, wherein the cell is isolated from a cell population selected from the group consisting of:

peripheral blood, umbilical cord blood, placenta, bone marrow, stem cell factor mobilized cells, and colony stimulating factor mobilized cells.

7. The isolated cell of claim 5, wherein the cell is isolated from amniotic fluid.

8. The isolated cell of claim 5, wherein the cell is of fetal origin within a maternal cell population.

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

i. selecting at least one stem cell according to claim 1, from a fetus using the DAZL specific marker;

ii. producing a display of the chromosomes of the embryo; and

iii. analyzing the displayed chromosomes.

10. The method of claim 9 wherein the at least one stem cell is derived from amniotic fluid.

11. A method of treating a disorder or a disease comprising administering to a patient in need thereof a stem cell according to claim 1 and providing conditions for differentiation of said cells into cells characterizing the tissue, thereby treating the individual suffering from the tissue disorder or disease.

12. The method of claim 11 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, fertility disease or disorder and heart disease or disorder.

13. The method of claim 11 which further comprises subjecting the somatic-derived stem cell to culturing conditions suitable for inducing cell proliferation, thereby obtaining an expanded stem cell population;

and introducing said 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.

14. The method of claim 11 which comprises direct administration of the stem cell into testis or ovaries thereby restoring fertility.

15. A method of isolating stem cells from a somatic source within an organism 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.

16. The method of claim 15 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.

17. The method of claim 15, wherein the molecular probe is a molecular beacon probe.

18. The method of claim 15, 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.

19. A kit for isolation, enrichment and detection of somatic stem cells within a specimen, said kit 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.

20. The kit of claim 19 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.

21. The kit of claim 19 further comprising reagents for genetic analysis of fetal and maternal cells.