Hydrogel-based vascular lineage cell growth media and uses thereof

Patent metadata

Field	Value
Publication number	US-9447381-B2
Application number	US-201414553442-A
Country	US
Kind code	B2
Filing date	Nov 25, 2014
Priority date	Nov 10, 2009
Publication date	Sep 20, 2016
Grant date	Sep 20, 2016

Abstract

A medium for growing vascular lineage cells is described. The vascular lineage cell growth medium includes an oligosaccharide-based hydrogel and a growth factor that promotes vascularization by vascular lineage cells.

First claim

We claim:

1. A method of preparing a vascular lineage cell growth medium comprising
chemically bonding an adhesion promoter to an oligosaccharide,
combining the chemically bonded adhesion promoter and oligosaccharide with at least one growth factor,
adding a crosslinking peptide moiety,
curing the combination to form a hydrogel, wherein the hydrogel has a Young's modulus between about 10 Pa and about 500 Pa; and
wherein the growth factor is an amount sufficient to promote vasculogenesis and tube formation, and sufficient to stimulate MMP production in vascular lineage cells.

2. The method of claim 1 , wherein the oligosaccharide is hyaluronic acid that is from about 20% to about 90% acrylate modified and is present in an amount of about 1 wt % to about 4 wt %.

3. The method of claim 1 , wherein the adhesion promoter is present at a concentration of from about 0.1 mM to about 20 mM.

4. The method of claim 1 , wherein the crosslinking peptide moiety is present at a concentration of from about 1 mM to about 8 mM.

5. The method of claim 1 , wherein the growth factor is present at a concentration of from about 25 ng/ml to about 200 ng/ml.

6. The method of claim 1 , wherein the hydrogel allows formation of capillary-like structures (CLSs) with extended vacuoles and open lumens from vascular lineage cells.

7. The method of claim 1 , wherein the hydrogel and crosslinking peptide moiety are cleavable by an enzyme.

8. The method of claim 7 , wherein the enzyme is a matrix metalloproteinase.

9. The method of claim 8 , wherein the matrix metalloproteinase is at least one of MMP-1, MMP-2 or MMP-10.

10. The method of claim 1 , wherein the crosslinking moiety peptide comprises the sequence SEQ ID NO: 1.

11. The method of claim 1 , wherein the adhesion promoter promotes vascular lineage cell adhesion to the hydrogel and promotes vacuole and lumen formation in vascular lineage cells.

12. The method of claim 1 , wherein the hydrogel has a Young's modulus between about 10 Pa and about 250 Pa.

13. The method of claim 1 , wherein the oligosaccharide is modified with an acryl group.

14. The method of claim 1 , wherein the oligosaccharide is acrylated hyaluronic acid.

15. The method of claim 1 , wherein the growth factor is selected from the group consisting of VEGF, TNFα, SDF1-α, bFGF, angiopoetin-1, PDGF, TGF-β, PIGF or combinations thereof.

16. The method of claim 1 , wherein the adhesion promoter is a peptide that includes an RGD sequence and a reactive functional group that is capable of bonding the adhesion promoter to the oligosaccharide.

17. A vascular lineage cell growth medium prepared by the method of claim 16 , wherein the oligosaccharide is acrylated hyaluronic acid.

18. A method of inducing vascular cell morphogenesis in vivo comprising contacting a subject with a vascular construct comprising the medium of claim 17 and vascular lineage cells.

19. The method of claim 18 , wherein the crosslinking moiety peptide comprises the sequence SEQ ID NO: 1.

20. The method of claim 18 , wherein the growth factor is selected from the group consisting of VEGF, TNFα, SDF1-α, bFGF, angiopoetin-1, PDGF, TGF-β, PIGF or combinations thereof.

21. The method of claim 1 , wherein the oligosaccharide is modified with a thiol group.

Assignees

• Univ Johns Hopkins

Inventors

• Gerecht Sharon
• Hanjaya-Putra Donny

Classifications

• C12N2501/148

  Transforming growth factor alpha [TGF-a] · CPC title

• C12N2501/165

  Vascular endothelial growth factor [VEGF] · CPC title

• C12N2533/50

  Proteins · CPC title

• A61K35/44

  Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells · CPC title

• A61L2300/414

  Growth factors · CPC title