USE OF MARKERS OF UNDIFFERENTIATED PLURIPOTENT STEM CELLS

• US 20130115623A1
• Filed: 03/10/2011
• Published: 05/09/2013
• Est. Priority Date: 03/10/2010
• Status: Active Grant

• Current Assignee: Agency For Science Technology and Research
• Original Assignee: Agency For Science Technology and Research
• Inventors: Choo, Boon Hwa Andre, Fong, Wey Jia
• Granted Patent: US 8,906,678 B2
• Time in Patent Office: Days
• US Class Current: 435/7.21
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07D 409/14 containing three or more hetero rings
• 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

Abstract

The disclosure relates to methods of binding and identifying undifferentiated pluripotent stem cells and particularly, although not exclusively, to use of binding moieties which bind to PHB on the surface of undifferentiated pluripotent stem cells, such as PHB-binding peptides, and to methods for depleting undifferentiated stem cells from a sample.

Claims

1. A method of identifying an undifferentiated pluripotent stem cell or cells that expresses PHB on their surface in a sample containing such cells, the method comprising identifying a cell or cells within the sample that express prohibitin (PHB) on their surface.

2. A method according to claim 1, wherein the method further comprises isolating an undifferentiated pluripotent stein cell or cells from a sample containing such cells, the method comprising isolating a cell or cells within the sample that express PHB on their surface.

3. A method according to claim 1, wherein the method further comprises removing an undifferentiated pluripotent stem cell or cells from a sample containing such cells, the method comprising removing from the sample a cell or cells that express PHB on their surface.

4. The method of claim 3, wherein the method comprises destroying an undifferentiated pluripotent stem cell or cells in a sample containing such cells, the method comprising destroying the cell or cells in the sample that express PHB on their surface.

5. A method according to claim 1 wherein the sample is contacted with a binding moiety which binds to PHB and the said undifferentiated pluripotent stem cell or cells are identified in the sample by virtue of being bound by the binding moiety.

6. A method according to claim 4 wherein the sample is contacted with a binding moiety which binds to PHB and the said undifferentiated pluripotent stem cell or cells are destroyed by virtue of being bound by the binding moiety.

7. A method according to claim 5 wherein the binding moiety is a peptide which has an amino acid sequence sharing at least 70% sequence identity with SEQ ID NO:

1.

8. A method according to claim 6 wherein the binding moiety is a peptide which has an amino acid sequence sharing at least 70% sequence identity with SEQ ID NO:

3.

9. A method of binding an undifferentiated pluripotent stem cell or cells in a sample containing one or a plurality of undifferentiated pluripotent stem cells, the method comprising:

providing a peptide which binds prohibitin (PHB); and

contacting the sample with the peptide under conditions permitting binding of the peptide to PHB on the surface of an undifferentiated pluripotent stein cell.

10. A method according to claim 9 further comprising:

identifying the undifferentiated pluripotent stem cell or cells by virtue of being bound to the peptide.

11. The method of claim 9 further comprising:

isolating the undifferentiated pluripotent stem cell or cells from the sample by separating cells bound to the peptide from cells not bound to the peptide.

20. The method of claim 10 further comprising:

isolating the undifferentiated pluripotent stem cell or cells from the sample by separating cells bound to the peptide from cells not bound to the peptide.

12. A method of enriching pluripotent stem cells that have undergone or are undergoing differentiation from a sample comprising undifferentiated pluripotent stem cells and pluripotent stem cells that have undergone or are undergoing differentiation, the method comprising:

(i) contacting the sample with a peptide which binds prohibitin (PHB) under conditions permitting the binding of the peptide to undifferentiated pluripotent stem cells; and

(ii) separating cells not bound to the peptide from cells bound to the peptide so as to generate a sample that is enriched in pluripotent stem cells that have undergone or are undergoing differentiation.

14. A method according to claim 12 wherein the cells bound to the peptide are destroyed by virtue of being bound by the peptide.

15. A method according to claim 14 wherein the peptide is a pro-apoptotic peptide.

13. A method of preparing a composition containing cells differentiated from undifferentiated pluripotent stem cells, which contains substantially no undifferentiated pluripotent stein cells, the method comprising:

(i) providing a population of cells comprising undifferentiated pluripotent stem cells and cells differentiated from undifferentiated pluripotent stem cells;

(ii) contacting the population with a peptide which binds prohibitin (PHB) under conditions permitting the binding of the peptide to undifferentiated pluripotent stein cells; and

(iii) separating cells not bound to the peptide from cells bound to the peptide.

21. A method according to claim 13 wherein the cells bound to the peptide are destroyed by virtue of being bound by the peptide.

22. A method according to claim 21 wherein the peptide is a pro-apoptotic peptide.

16. (canceled)

17. A method of destroying an undifferentiated pluripotent stem cell or cells in a sample containing such cells, the method comprising:

(i) contacting the sample with a pro-apoptotic peptide which binds prohibitin (PHB) under conditions permitting the binding of the peptide to undifferentiated pluripotent stein cells; and

(ii) allowing the peptide to destroy the said cell.

18. A method according to claim 17 wherein the peptide has an amino acid sequence sharing at least 70% sequence identity with SEQ ID NO:

1.

19. A method according to claim 17 wherein the peptide has an amino acid sequence sharing at least 70% sequence identity with SEQ ID NO 3.