Artificial three-dimensional microenvironment niche culture and methods of using the same

• US 9,988,607 B2
• Filed: 06/28/2017
• Issued: 06/05/2018
• Est. Priority Date: 12/13/2008
• Status: Active Grant

• Current Assignee: Babak Esmaeli-Azad
• Original Assignee: Babak Esmaeli-Azad
• Inventors: Esmaeli-Azad, Babak
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US15/636,548
• Publication Number:
  US 20170306297A1
• Time in Patent Office: 342 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 signal transduction pathways, wherein the one or more signal transduction pathways is selected from the group consisting of Wnt, Hedgehog, Cyclooxygenase, p53, NF-KB, PKC, PGE-2 and combinations thereof;

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 chemicals comprises 6-bromoindirubin-3′

-oxime (BIO).

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

-oxime (INO).

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

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

).

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

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

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

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

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

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

12. The artificial three-dimensional microenvironment niche culture of claim 1, wherein the valproic acid concentration is about 0.02 mM to 10 mM.

13. The artificial three-dimensional microenvironment niche culture of claim 1, wherein the synthetic polymer hydrogel comprises hyaluronan.

14. The artificial three-dimensional microenvironment niche culture of claim 1, wherein the at least one growth factor is selected from the group consisting of epidermal growth factor, fibroblast growth factor, noggin, and combinations thereof.