ARTIFICIAL THREE-DIMENSIONAL MICROENVIRONMENT NICHE CULTURE AND METHODS OF USING THE SAME

• US 20170306297A1
• Filed: 06/28/2017
• Published: 10/26/2017
• Est. Priority Date: 12/13/2008
• Status: Active Grant

• Current Assignee: DNAmicroarray, Inc.
• Original Assignee: DNAmicroarray, Inc.
• Inventors: Esmaeli-Azad, Babak
• Granted Patent: US 9,988,607 B2
• Time in Patent Office: Days
• CPC Class Codes: C12N 2501/02 Compounds of the arachidonic acid pathway, e.g. prostaglandins, leukotrienes
• CPC Class Codes: C12N 2501/065 Modulators of histone acetylation
• CPC Class Codes: C12N 2501/415 Wnt; Frizzeled
• CPC Class Codes: C12N 2503/00 Use of cells in diagnostics
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2539/00 Supports and/or coatings for cell culture characterised by properties
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS
• CPC Class Codes: G01N 2500/10 involving cells
• CPC Class Codes: G01N 33/5073 Stem cells

Abstract

Described here are three-dimensional microenvironment niches prepared from biomaterial compositions that support growth and self renewal of stem cells. The invention also provides methods for inducing pluripotency in a somatic cell using chemical compounds, as well as methods for screening for compounds that can induce pluripotency in a somatic cell.

Claims

1. An artificial three-dimensional microenvironment niche culture comprising:

a) a synthetic polymer hydrogel;

b) a culture medium;

c) one or more chemicals which target or affect one or more iPS signal transduction pathways;

d) at least one growth factor;

e) a cell; and

f) valproic acid.

2. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include Wnt pathways.

3. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include Hedgehog pathways.

4. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include Cyclooxygenase pathways.

5. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include p53 pathways.

6. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include NF-KB pathways.

7. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include PKC pathways.

8. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more iPS signal transduction pathways include PGE-2 pathways.

9. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more chemicals includes 6-bromoindirubin-3′

-oxime (BIO).

10. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more chemicals includes indirubin-5-nitro-3′

-oxime (INO).

11. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more chemicals includes 2-(3-(6-Methylpyridin-2-yl)-1H-pyrazol-4-yl)-1,5-napthyridine.

12. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more chemicals includes 1-4(-Methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl)ethanone HBr (Pifithrin-α

).

13. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said one or more chemicals includes prostaglandin J2.

14. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises laminin.

15. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises fibronectin.

16. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises vitronectin.

17. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises epidermal growth factor.

18. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises fibroblast growth factor.

19. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises Noggin.

20. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises SIS.

21. The artificial three-dimensional microenvironment niche culture of claim 1, wherein said synthetic polymer hydrogel further comprises EHS basement membrane.