METHODS FOR ISOLATING VERY SMALL EMBRYONIC-LIKE (VSEL) STEM CELLS

• US 20100267107A1
• Filed: 09/30/2009
• Published: 10/21/2010
• Est. Priority Date: 10/30/2007
• Status: Abandoned Application

• Current Assignee: University of Louisville
• Original Assignee: University of Louisville
• Inventors: Zuba-Surma, Ewa K., Kucia, Magdalena, Ratajczak, Mariusz, Ratajczak, Janina
• Application Number:
  US12/570,181
• Publication Number:
  US 20100267107A1
• Time in Patent Office: Days
• US Class Current: 435/173.900
• 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: A61K 38/193 Colony stimulating factors [CSF]
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696

Abstract

The presently disclosed subject matter provides methods of isolating populations of stem cells that from bone marrow, peripheral blood, and/or other sources. Also provided are methods of using the stem cells for treating tissue and/or organ damage in a subject.

Claims

1. An enriched population of very small embryonic like stem cells (VSELs) derived from adult organ or tissue cells of a human, wherein the population is enriched by selecting cells for CD133⁺ CXCR4⁺ CD34⁺ Lin⁻

CD45⁻

cells to obtain an enriched population of target VSELs.

2. The enriched population of VSELs of claim 1, wherein the target VSELs are Oct-4⁺.

3. The enriched population of VSELs of claim 1, wherein the target VSELs are SSEA⁺.

4. The enriched population of VSELs of claim 1, wherein the target VSELs express Oct-4 protein in nuclei and SSEA antigens on the surface.

5. The enriched population of VSELs of claim 1, wherein the target VSELs are Nanog⁺.

6. The enriched population of VSELs of claim 1, wherein the target VSELs express markers of primordial germ cells (PGCs) selected from the group consisting of fetal-type alkaline phosphatase, OctA, SSEA-1, CXCR4, Mvh, Stella, Fragilis, Nobox and Hdac6.

7. The enriched population of VSELs of claim 1, wherein the population is enriched by selecting for cells that are 2 to 6 μ

m in size.

8. The enriched population of VSELs of claim 1, wherein the population is enriched by selecting for cells that are 2 to 4 μ

m in size.

9. The enriched population of VSELs of claim 1, wherein the population is enriched by selecting for cells that contain primitive unorganized euchromatin.

10. The enriched population of VSELs of claim 1, wherein at least 30% of the cells in the population are target VSELs.

11. An enriched population of very small embryonic like stem cells (VSELs) derived from the blood a human, wherein the population is enriched by selecting cells for CD133⁺ CXCR4⁺ CD34⁺ Lin⁻

CD45⁻

cells to obtain an enriched population of target VSELs.

12. The enriched population of VSELs of claim 11, wherein the blood is cord blood.

13. The enriched population of VSELs of claim 11, wherein the blood is peripheral blood.

14. The enriched population of VSELs of claim 11, wherein the target VSELs are Oct-4⁺.

15. The enriched population of VSELs of claim 11, wherein the target VSELs are SSEA⁺.

16. The enriched population of VSELs of claim 11, wherein the target VSELs express Oct-4 protein in nuclei and SSEA antigens on the surface.

17. The enriched population of VSELs of claim 11, wherein the target VSELs are Nanog⁺.

18. The enriched population of VSELs of claim 11, wherein the target VSELs express markers of primordial germ cells (PGCs) selected from the group consisting of fetal-type alkaline phosphatase, OctA, SSEA-1, CXCR4, Mvh, Stella, Fragilis, Nobox and Hdac6.

19. The enriched population of VSELs of claim 11, wherein the population is enriched by selecting for cells that are 2 to 6 μ

m in size.

20. The enriched population of VSELs of claim 11, wherein the population is enriched by selecting for cells that are 2 to 4 μ

m in size.

21. The enriched population of VSELs of claim 11, wherein the population is enriched by selecting for cells that contain primitive unorganized euchromatin.

22. The enriched population of VSELs of claim 11, wherein at least 30% of the cells in the population are target VSELs.

23. A method of producing population of cells enriched for target very small embryonic like stem cells (VSELs) from the blood of a human, comprising lysing the blood to deplete erythrocytes and preparing an enriched population of Lin⁻

/CD45⁻

/CD133⁺ cells.

24. The method of claim 23, wherein the blood is cord blood.

25. The method of claim 23, wherein the blood is peripheral blood.

26. A method of producing population of cells enriched for target very small embryonic like stem cells (VSELs) from the blood of a human, comprising i) lysing the blood to deplete erythrocytes;

ii) preparing an enriched population of CD133⁺ cells; and

iii) preparing an enriched population of Lin⁻

/CD45⁻

/CD133⁺.

27. The method of claim 26, wherein the blood is cord blood.

28. The method of claim 26, wherein the blood is peripheral blood.

29. The method of claim 26 wherein the enriched population of Lin⁻

/CD45⁻

/CD133⁺ are enriched by fluorescence-activated cell sorting.

30. The method of claim 26 wherein the blood is lysed in a hypotonic ammonium chloride solution to deplete erythrocytes.

31. The method of claim 26 wherein the CD133⁺ cells are enriched by employing immunomagnetic beads.