IP Library Granted Patent US 9,683,213
Granted Patent B2
US 9,683,213 · App. 14/684,120 · Granted Jun 20, 2017

Hydrogel compositions for use in cell expansion and differentiation

Inventors: William L. Murphy (Waunakee, WI); Matthew Brian Parlato (Madison, WI); James A Molenda (Madison, WI); Ngoc Nhi Le (Norcross, GA)
Assignee: Wisconsin Alumni Research Foundation
C12N5/0606G01N33/4833B01J19/0046B01J2219/00596C12N2533/20C12N2533/30C12N2537/10G01N21/6452G01N2610/00
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Quick Facts
Patent No.
US 9,683,213
App. No.
14/684,120
Granted
Jun 20, 2017
Kind
B2
Abstract

Hydrogel compositions and methods of using hydrogel compositions are disclosed. Advantageously, the hydrogel compositions offer the ability to promote cellular expansion and/or cellular differentiation of various cells.

Claims (28)

1. A method of promoting cellular expansion, the method comprising:

preparing a hydrogel composition, wherein the hydrogel composition comprises an 8-arm, 20 kDa polyethylene glycol functionalized with norbornene, a crosslinking peptide, and a cell adhesion peptide;

contacting a cell with the hydrogel composition; and

culturing the cell.

2. The method of claim 1 wherein the cell is a circulating angiogenic cell.

3. The method of claim 2 wherein the hydrogel composition comprises at least 1 mM cell adhesion peptide selected from the group consisting of CRGDS (SEQ ID NO:2), Acetylated-GCYGRGDSPG (SEQ ID NO:31), cyclic RGD (SEQ ID NO:35), CRGD-(G) 13 -PHSRN (SEQ ID NO:29), CPHSRN-(SG) 5 -RGD (SEQ ID NO:30) and IKVAV (SEQ ID NO:38).

4. The method of claim 2 wherein the hydrogel composition comprises a shear modulus range of from about 2 kPa to about 12 kPa.

5. The method of claim 1 wherein the cell is a human mesenchymal stem cell.

6. The method of claim 5 wherein the hydrogel composition comprises a shear modulus of from about 1.8 kPa to about 33 kPa.

7. The method of claim 5 wherein the hydrogel composition comprises at least 0.25 mM of cell adhesion peptide selected from the group consisting of CRGDS (SEQ ID NO:2), Acetylated-GCYGRGDSPG (SEQ ID NO:31), cyclic RGD (SEQ ID NO:35), CRGD-(G) 13 -PHSRN (SEQ ID NO:29), CPHSRN-(SG) 5 -RGD (SEQ ID NO:30) and IKVAV (SEQ ID NO:38).

8. The method of claim 5 wherein the cell is a human pluripotent stem cell selected from the group consisting of a human embryonic stem cell and human induced pluripotent stem cell.

9. The method of claim 8 wherein the hydrogel composition comprises at least 0.25 mM of cell adhesion peptide selected from the group consisting of CRGDS (SEQ ID NO:2), Acetylated-GCYGRGDSPG (SEQ ID NO:31), cyclic RGD (SEQ ID NO:35), CRGD-(G) 13 -PHSRN (SEQ ID NO:29), CPHSRN-(SG) 5 -RGD (SEQ ID NO:30) and IKVAV (SEQ ID NO:38).

10. The method of claim 8 wherein the hydrogel composition comprises a shear modulus of from about 3 kPa to about 16 kPa.

11. The method of claim 8 wherein the hydrogel composition further comprises immobilized low molecular weight heparin.

12. A method of promoting cellular differentiation, the method comprising:

preparing a hydrogel composition, wherein the hydrogel composition comprises an 8-arm, 20 kDa polyethylene glycol functionalized with norbornene, a crosslinking peptide, and a cell adhesion peptide;

contacting a cell with the hydrogel composition; and

culturing the cell.

13. The method of claim 12 wherein the cell is a human mesenchymal stem cell.

14. The method of claim 13 wherein the hydrogel composition comprises a shear modulus of from about 1.8 kPa to about 33 kPa.

15. The method of claim 13 wherein the hydrogel composition comprises at least 0.25 mM of cell adhesion peptide selected from the group consisting of CRGDS (SEQ ID NO:2), Acetylated-GCYGRGDSPG (SEQ ID NO:31), cyclic RGD (SEQ ID NO:35), CRGD-(G) 13 -PHSRN (SEQ ID NO:29), CPHSRN-(SG) 5 -RGD (SEQ ID NO:30) and IKVAV (SEQ ID NO:38).

16. The method of claim 12 wherein the cell is a human pluripotent stem cell selected from the group consisting of a human embryonic stem cell and human induced pluripotent stem cell.

17. The method of claim 16 wherein the hydrogel composition comprises at least 0.25 mM of cell adhesion peptide selected from the group consisting of CRGDS (SEQ ID NO:2), Acetylated-GCYGRGDSPG (SEQ ID NO:31), cyclic RGD (SEQ ID NO:35), CRGD-(G) 13 -PHSRN (SEQ ID NO:29), CPHSRN-(SG) 5 -RGD (SEQ ID NO:30) and IKVAV (SEQ ID NO:38).

18. The method of claim 16 wherein the hydrogel composition comprises a shear modulus of from about 3 kPa to about 16 kPa.

19. A method of promoting cellular expansion, the method comprising:

preparing a hydrogel composition, wherein the hydrogel composition comprises a polyethylene glycol functionalized with norbornene, a crosslinking peptide, a cell adhesion peptide, and immobilized low molecular weight heparin;

contacting a cell with the hydrogel composition; and

culturing the cell.

Assignments (2)
CONFIRMATORY LICENSE Recorded May 17, 2018
From: WISCONSIN ALUMNI RESEARCH FOUNDATION
To: NATIONAL INSTITUTES OF HEALTH (NIH), U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS), U.S. GOVERNMENT
Reel/Frame 046179/0744 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 7, 2015
From: MURPHY, WILLIAM; PARLATO, MATTHEW; LE, NGOC NHI; MOLENDA, JAMES
To: WISCONSIN ALUMNI RESEARCH FOUNDATION
Reel/Frame 036275/0749 →
Continuity (2)
Provisional Application 61978032 · Apr 10, 2014
Related Publication 20150291930A1 · Oct 15, 2015