Three dimensional matrix for cancer stem cells
Synthetic inert 3D gel culture systems are described that can be finely tuned to exhibit desired and predetermined physical, chemical, mechanical, and biochemical properties. The culture system can be utilized to study the effect of microenvironmental factors on cancer cell response, and in particular on cancer stem cell (CSC) response. Cancer cells can be encapsulated in a crosslinked gel system having a narrow range of predetermined gel stiffness. One or more biochemical factors including peptides that can affect the growth, development, and/or proliferation of CSCs can be incorporated in the system to examine the effects of the factor(s) on the encapsulated cells with regard to growth, proliferation, size, etc.
1. A method of forming a three dimensional hydrogel matrix for supporting a cancer cell, the method comprising:
combining an inert polyethylene glycol diacrylate homopolymer with a crosslinking agent to form a precursor solution;
crosslinking the inert polyethylene glycol diacrylate homopolymer via reaction between the diacrylate of the homopolymer and the crosslinking agent to form an inert three dimensional hydrogel matrix that is absent of ligands that can interact with cell surface receptors, wherein the concentration of the crosslinking agent and/or the concentration of the inert polyethylene glycol diacrylate homopolymer is predetermined in the precursor solution such that the inert three dimensional hydrogel matrix has a predetermined elastic modulus; and
following formation of the inert three dimensional hydrogel matrix, conjugating a peptide to the matrix, the peptide affecting the growth, development, and/or proliferation of a cancer stem cell.
2. The method of claim 1 , further comprising encapsulating a population of cells in the three dimensional hydrogel matrix, the population of cells comprising cancer stem cells.
3. The method of claim 2 , the population of cells further comprising differentiated cancer cells.
4. The method of claim 3 , wherein the differentiated cancer cells comprise breast cancer cells.
5. The method of claim 2 , the population of cells further comprising tumor stroma cells.
6. The method of claim 1 , further comprising combining the inert polyethylene glycol diacrylate homopolymer and the crosslinking agent with an initiator.
7. The method of claim 6 , further comprising subjecting the inert polyethylene glycol diacrylate homopolymer to radiation to initiate the crosslinking.
8. The method of claim 1 , wherein the precursor solution includes the inert polyethylene glycol diacrylate homopolymer in a concentration of about 10% by weight of the precursor solution or less and the predetermined elastic modulus is about 10 kilopascals or less.
9. The method of claim 1 , wherein the precursor solution includes the inert polyethylene glycol diacrylate homopolymer in a concentration of from about 10% by weight of the precursor solution to about 20% by weight of the precursor solution and the predetermined elastic modulus is from about 10 kilopascals to about 30 kilopascals.
10. The method of claim 1 , wherein the precursor solution includes the inert polyethylene glycol diacrylate homopolymer in a concentration of about 20% by weight of the precursor solution or greater and the predetermined elastic modulus is about 30 kilopascals or greater.
11. The method of claim 1 , wherein the peptide is a CD44 binding peptide or a mutant thereof, an integrin binding peptide or a mutant thereof, or a heparain binding peptide or a mutant thereof.
12. The method of claim 1 , wherein the peptide is RLVSYNGIIFFLK (SEQ ID NO.: 17), VLFGFLKIYSRIN (SEQ ID NO.: 18), GRGDS (SEQ ID NO.: 19), GRDGS (SEQ ID NO.: 20), WQPPRARI (SEQ ID NO.: 21), or RPQIPWAR (SEQ ID NO.: 22).
13. The method of claim 1 further comprising incorporating a biochemical factor in the hydrogel matrix.
14. A method of forming a three dimensional hydrogel matrix for supporting a cancer cell, the method comprising:
combining an inert polyethylene glycol diacrylate homopolymer with a crosslinking agent to form a precursor solution;
crosslinking the inert polyethylene glycol diacrylate homopolymer via reaction between the diacrylate of the homopolymer and the crosslinking agent to form an inert three dimensional hydrogel matrix that is absent of ligands that can interact with cell surface receptors, wherein the concentration of the crosslinking agent and/or the concentration of the inert polyethylene glycol diacrylate homopolymer is predetermined in the precursor solution such that the inert three dimensional hydrogel matrix has a predetermined elastic modulus; and
encapsulating a population of cells in the inert three dimensional hydrogel matrix, the population of cells comprising cancer stem cells.
15. The method of claim 14 , the population of cells further comprising differentiated cancer cells.
16. The method of claim 15 , the differentiated cancer cells comprising breast cancer cells.
17. The method of claim 14 , the population of cells further comprising tumor stroma cells.
18. The method of claim 14 , further comprising combining the inert polyethylene glycol diacrylate homopolymer and the crosslinking agent with an initiator.
19. The method of claim 18 , further comprising subjecting the inert polyethylene glycol diacrylate homopolymer to radiation to initiate the crosslinking.
20. The method of claim 14 , wherein the precursor solution includes the inert polyethylene glycol diacrylate homopolymer in a concentration of about 10% by weight of the precursor solution or less and the predetermined elastic modulus is about 10 kilopascals or less.
21. The method of claim 14 , wherein the precursor solution includes the inert polyethylene glycol diacrylate homopolymer in a concentration of from about 10% by weight of the precursor solution to about 20% by weight of the precursor solution and the predetermined elastic modulus is from about 10 kilopascals to about 30 kilopascals.
22. The method of claim 14 , wherein the precursor solution includes the inert polyethylene glycol diacrylate homopolymer in a concentration of about 20% by weight of the precursor solution or greater and the predetermined elastic modulus is about 30 kilopascals or greater.
23. The method of claim 14 , further comprising following formation of the inert three dimensional hydrogel matrix, conjugating a peptide to the inert matrix, wherein the peptide is a CD44 binding peptide or a mutant thereof, an integrin binding peptide or a mutant thereof, or a heparain binding peptide or a mutant thereof.
24. The method of claim 14 , further comprising following formation of the inert three dimensional hydrogel matrix, conjugating a peptide to the inert matrix, wherein the peptide is RLVSYNGIIFFLK (SEQ ID NO.: 17), VLFGFLKIYSRIN (SEQ ID NO.: 18), GRGDS (SEQ ID NO.: 19), GRDGS (SEQ ID NO.: 20), WQPPRARI (SEQ ID NO.: 21), or RPQIPWAR (SEQ ID NO.: 22).
25. The method of claim 14 , further comprising incorporating a biochemical factor in the hydrogel matrix.