Method for acceleration of stem cell differentiation

• US 20070048865A1
• Filed: 03/15/2006
• Published: 03/01/2007
• Est. Priority Date: 03/15/2005
• Status: Abandoned Application

• Current Assignee: The Central Institute for Experimental Animals
• Original Assignee: The Central Institute for Experimental Animals
• Inventors: Kurita, Ryo, Tani, Kenzaburo
• Application Number:
  US11/377,847
• Publication Number:
  US 20070048865A1
• Time in Patent Office: Days
• US Class Current: 435/456
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

The present invention relates to methods for accelerating the differentiation of embryonic stem cells by introducing nucleic acid encoding the Tal1/Scl transcription factor into the cells, and culturing the transduced cells in a differentiation medium.

Claims

1. A method for accelerating differentiation of embryonic stem cells, comprising introducing nucleic acid encoding the Tal1/Scl transcription factor into embryoid bodies formed from undifferentiated embryonic stem cells, and culturing the embryoid bodies carrying the Tal1/Scl encoding nucleic acid in a differentiation medium until differentiation is confirmed.

2. The method of claim 1 wherein said differentiation is hematopoietic differentiation.

3. The method of claim 2 wherein said nucleic acid is introduced into said embryoid bodies by vector-mediated gene transfer.

9. The method of claim 2 wherein differentiation is confirmed by confirming the presence of multilineage blood cells.

10. The method of claim 9 wherein the presence of multilineage blood cells is confirmed by an immunochemical test.

11. The method of claim 9 wherein the presence of multilineage blood cells is confirmed by morphological analysis.

12. The method of claim 2 wherein differentiation is confirmed by detecting regional expression of at least one embryonic marker specific for a cellular lineage.

13. The method of claim 12 wherein said embryonic marker is selected from the group consisting of zeta-globin, neurofilament 68 kD, and alpha-fetoprotein.

14. The method of claim 2 wherein said differentiation medium comprises one or more cytokines and/or growth factors.

15. The method of claim 2 wherein said differentiation medium comprises one or more ingredients selected from the group consisting of IL-3, IL-6, GM-CSF, G-CSF, SCF, and erythropoietin.

16. The method of claim 15 wherein said differentiation medium comprises all of said ingredients.

17. The method of claim 1 wherein said embryonic stem cell is a primate cell.

18. The method of claim 17 wherein said primate is a non-human primate.

19. The method of claim 18 wherein said non-human primate is a marmoset.

20. The method of claim 17 wherein said primate is a human.

4. The method of clam 3 wherein said vector is a viral vector.

5. The method of claim 4 wherein said vector is a lentiviral vector.

6. The method of claim 5 wherein said lentiviral vector is VSV-G pseudotyped.

7. The method of claim 6 wherein said lentiviral vector comprises the Tal1/Scl gene under control of a promoter selected from the group consisting of EF-1a, CAG, PGK and CMV promoters.

8. The method of claim 7 wherein said promoter is an EF-1a promoter.