Three-dimensional cell adhesion matrix

Patent metadata

Field	Value
Publication number	US-9296988-B2
Application number	US-201013378540-A
Country	US
Kind code	B2
Filing date	Jun 18, 2010
Priority date	Jun 19, 2009
Publication date	Mar 29, 2016
Grant date	Mar 29, 2016

Abstract

The present invention provides a cell adhesion matrix having poly(vinyl alcohol) chains crosslinked via carboxy phenyl boronic acid crosslinkers. The cell adhesion matrix can also include a molecular recognition element bound to the poly(vinyl alcohol) chains via a carboxy phenyl boronic acid group, as well as including cells. The present invention also provides a method for making and de-gelling the cell adhesion matrix.

First claim

What is claimed is:

1. A cell adhesion matrix prepared by the process comprising:
contacting a plurality of poly(vinyl alcohol) chains and at least one boronic acid crosslinker having at least two boronic acids in a mixture, under conditions such that each boronic acid becomes linked to two adjacent hydroxy groups of one of the plurality of poly(vinyl alcohol) chains,
wherein
the boronic acid crosslinker is of formula I:

wherein
Linker comprises a biocompatible polymer selected from the group consisting of a poly(propylene glycol) polymer and a poly(ethylene glycol) polymer of the following formula:

wherein
X 1 and X 2 are each independently selected from the group consisting of a bond, a branching moiety, lysine and a lysine derivative; and
subscript p is from about 10 to about 500;

subscript m is from about 1 to about 100; and

each poly(vinyl alcohol) shain is of formula II:

wherein subscript n is from about 10 to about 5000.

2. The cell adhesion matrix of claim 1 , wherein the mixture further comprises a plurality of cells.

3. The cell adhesion matrix of claim 1 , wherein the conditions include a pH of from about 7.0 to about 8.0.

4. The cell adhesion matrix of claim 1 , wherein X 1 and X 2 are each a bond.

5. The cell adhesion matrix of claim 1 , wherein X 1 and X 2 are each lysine.

6. The cell adhesion matrix of claim 1 , wherein subscript p is from about 40 to about 50.

7. The cell adhesion matrix of claim 1 , wherein the boronic acid crosslinker is selected from the group consisting of:

wherein subscript p is from about 40 to about 50, subscripts x, y and z are each independently from 1 to about 75 such that x+y+z is from about 80 to about 90, and subscripts a, b and c are each independently from about 10 to about 100, such that a+b+c is from about 100 to about 200, at a pH of about 7.4 such that each boronic acid becomes linked to two adjacent hydroxy groups of one of the plurality of modified poly(vinyl alcohol) chains.

8. The cell adhesion matrix of claim 1 , the process further comprising:
contacting a molecular recognition element conjugate and the plurality of poly(vinyl alcohol) chains in a mixture, wherein the molecular recognition element conjugate comprises a molecular recognition element and at least one binding group, under conditions such that each binding group of the molecular recognition element conjugate becomes linked to one of the poly(vinyl alcohol) chains.

9. The cell adhesion matrix of claim 8 , wherein the molecular recognition element conjugate has the following formula III:

10. The cell adhesion matrix of claim 8 , wherein the molecular recognition element is selected from the group consisting of LXY3, MSE, HYD1, LLP-2A, and RGD ligand.

11. The cell adhesion matrix of claim 1 , wherein the poly(vinyl alcohol) is linked to at least one molecular recognition element.

12. A cell adhesion matrix comprising
at least one boronic acid crosslinker of formula I:

wherein
Linker comprises a biocompatible polymer selected from the group consisting of a poly(propylene glycol) polymer and a poly(ethylene glycol) polymer of the following formula:

wherein
X 1 and X 2 are each independently selected from the group consisting of a bond, a branching moiety, lysine and a lysine derivative; and
subscript p is from about 10 to about 500;

subscript m is from 1 to about 100; and

a plurality of poly(vinyl alcohol) chains each of formula II:

wherein subscript n is from about 10 to about 5000,

wherein each boronic acid is linked to two adjacent hydroxy groups of one of the plurality of poly(vinyl alcohol) chains.

13. The cell adhesion matrix of claim 12 , wherein the cell adhesion matrix further comprises a molecular recognition element conjugate.

14. The cell adhesion matrix of claim 12 , wherein the cell adhesion matrix further comprises a cell.

15. A cell adhesion matrix prepared by the process comprising:
contacting a molecular recognition element conjugate of formula (III):

wherein the molecular recognition element is selected from the group consisting of LXY3, MSE, HYD1, LLP-2A, and RGD ligand, and a plurality of poly(vinyl alcohol) chains of formula (II):

wherein subscript n is from about 10 to about 5000,

at a pH of about 7.4 such that each boronic acid of the molecular recognition element conjugate becomes linked to two adjacent hydroxy groups of one of the poly(vinyl alcohol) chains to form a plurality of modified poly(vinyl alcohol) chains; and

contacting the plurality of modified poly(vinyl alcohol) chains and at least one boronic acid crosslinker selected from the group consisting of:

wherein subscript p is from about 40 to about 50, subscripts x, y and z are each independently from 1 to about 75 such that x+y+z is from about 80 to about 90, and subscripts a, b and c are each independently from about 10 to about 100, such that a+b+c is from about 100 to about 200, at a pH of about 7.4 such that each boronic acid becomes linked to two adjacent hydroxy groups of one of the plurality of modified poly(vinyl alcohol) chains.

Assignees

• Onofiok Ekama
• Lam Kit S
• Luo Juntao
• Univ California

Inventors

• Onofiok Ekama
• Lam Kit S
• Luo Juntao

Classifications

• C09J129/04 Primary

  Polyvinyl alcohol; Partially hydrolysed homopolymers or copolymers of esters of unsaturated alcohols with saturated carboxylic acids · CPC title

• C12N5/0062 Primary

  General methods for three-dimensional culture · CPC title

• C08K5/55

  Boron-containing compounds {(C08K5/0091 takes precedence)} · CPC title

• C12N2533/30

  Synthetic polymers · CPC title