Growth factor sequestering and presenting hydrogels

Patent metadata

Field	Value
Publication number	US-9827272-B2
Application number	US-201414760141-A
Country	US
Kind code	B2
Filing date	Jan 16, 2014
Priority date	Jan 17, 2013
Publication date	Nov 28, 2017
Grant date	Nov 28, 2017

Abstract

Provided herein are hydrogel cell matrices, hydrogel cell matrix systems for the support, growth, and differentiation of a stem cell or progenitor cell and methods for making such hydrogel cell matrices.

First claim

What is claimed is:

1. A hydrogel cell matrix comprising:
a) a plurality of hydrogel polymers;
b) a cell adhesion peptide conjugated to one or more hydrogel polymers;
c) a factor release molecule conjugated to one or more hydrogel polymers, wherein the factor release molecule is present in the matrix at a density of 0.01 weight % to 0.03 weight %; and
d) a proteolytically cleavable cross-linker peptide that links one or more hydrogel polymers of the plurality of hydrogel polymers to another hydrogel polymer of the plurality, wherein the cross-linker peptide is CQPQGLAKC (SEQ ID NO:46), and
wherein the matrix has an elastic modulus of from about 15 Pa to 850 Pa.

2. The hydrogel cell matrix of claim 1 , wherein the hydrogel polymer is a hyaluronic acid (HyA) polymer or an acrylated hyaluronic acid (HyA) polymer.

3. The hydrogel cell matrix of claim 1 , wherein the cell adhesion peptide and the factor release molecule are conjugated to the same hydrogel polymer.

4. The hydrogel cell matrix of claim 1 , wherein the cell adhesion peptide and the factor release molecule are conjugated to different hydrogel polymers.

5. The hydrogel cell matrix of claim 1 , wherein the cell adhesion peptide comprises the amino acid sequence RGD.

6. The hydrogel cell matrix of claim 5 , wherein the cell adhesion peptide comprises the amino acid sequence CGGNGEPRGDTYRAY (SEQ ID NO:1).

7. The hydrogel cell matrix of claim 1 , wherein the factor release molecule is heparin.

8. The hydrogel cell matrix of claim 1 , further comprising a growth factor non-covalently bound to the factor release molecule, wherein the factor release molecule allows for the release, presentation, or release and presentation of the growth factor.

9. The hydrogel cell matrix of claim 8 , wherein the growth factor is transforming growth factor-β (TGF-β).

10. The hydrogel cell matrix of claim 8 , wherein the growth factor was secreted by a cell cultured on the hydrogel cell matrix.

11. The hydrogel cell matrix of claim 1 , wherein the cross-linker is a thiolated cross linker peptide.

12. A hydrogel cell matrix system comprising the hydrogel cell matrix of claim 1 and a stem cell or progenitor cell encapsulated in the matrix.

13. The hydrogel cell matrix system of claim 12 , wherein the system comprises a progenitor cell.

14. The hydrogel cell matrix system of claim 13 , wherein the progenitor cell is a cardiac progenitor cell.

15. A method of making a hydrogel cell matrix comprising the steps of:
(a) conjugating cell adhesion peptides to a first population of hydrogel polymers;
(b) conjugating factor release molecules to a second population of hydrogel polymers; and
(c) conjugating the first population of hydrogel polymers to the second population of hydrogel polymers using proteolytically cleavable cross-linker peptides,
thereby producing a hydrogel cell matrix.

16. The method of claim 15 , further comprising, after step (b), a step of contacting the factor release molecules with a growth factor, wherein the growth factor non-covalently binds to the factor release molecules, thereby producing a growth factor contacted hydrogel cell matrix.

17. The method of claim 16 , further comprising culturing a cell on the growth factor contacted hydrogel cell matrix, wherein factors secreted by the cell non-covalently bind to the factor release molecules.

18. The method of claim 15 , further comprising, after step (b), culturing a cell in the presence of the factor release molecules, wherein factors secreted by the cell non-covalently bind to the factor release molecules.

19. The method of claim 15 , further comprising, after step (c), culturing a cell on the hydrogel cell matrix, wherein factors secreted by the cell non-covalently bind to the factor release molecules.

20. A method of introducing a hydrogel cell matrix into a mammalian subject, comprising, prior to gelation, injecting the hydrogel cell matrix of claim 1 into the mammalian subject.

21. The method of claim 20 , wherein the hydrogel cell matrix comprises a growth factor non-covalently bound to the factor release molecule, wherein the factor release molecule allows for the release, presentation, or release and presentation of the growth factor.

22. The method of claim 21 , wherein the growth factor was secreted by a cell cultured on the hydrogel cell matrix.

23. The method of claim 20 , wherein the hydrogel cell matrix comprises a stem cell or progenitor cell.

24. The hydrogel cell matrix of claim 1 , wherein the hydrogel polymer is present in the matrix at a density of from 1 weight % to 3 weight %.

25. The hydrogel cell matrix of claim 1 , wherein the crosslinking density is from about 25% to 100%.

Assignees

• Univ California

Inventors

• Jha Amit Kumar
• Healy Kevin E

Classifications

• C12N2533/70

  Polysaccharides · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• A61K47/48784

  Human Necessities · mapped topic

• C12N2501/91

  Heparin · CPC title

• A61K47/48338

  Human Necessities · mapped topic