Modified reconstituted basement membrane composition for assay system
View Patent ↗A modified reconstituted extracellular matrix composition is provided herein. The composition includes an extracellular matrix and at least one exogenous component selected from heparin, fibronectin and laminin. The composition may have a basic pH. Additionally, provided herein is a cell culturing system including a substrate and a coating thereon of the composition to assess potential stimulators and/or inhibitors for their effects on various cell cultures while increasing the signal dynamic range.
1. A method of preparing a cell-culturing system comprising:
(a) providing an extracellular matrix composition comprising a growth factor reduced extracellular matrix extracted from Engelbreth-Holm-Swarm mouse sarcoma tumor; and at least one exogenous component selected from the group consisting of heparin, fibronectin and laminin;
(b) applying the composition to a substrate; and
(c) incubating the applied composition to allow polymerization thereof.
2. The method of claim 1 , wherein the at least one exogenous component comprises heparin.
3. The method of claim 2 , further comprising laminin.
4. The method of claim 2 , further comprising fibronectin.
5. The method of claim 4 , further comprising laminin.
6. The method of claim 1 , wherein the at least one exogenous component comprises fibronectin.
7. The method of claim 6 , further comprising laminin.
8. The method of claim 1 , wherein the at least one exogenous component comprises laminin.
9. The method of claim 1 , wherein the composition has a basic pH.
10. The method of claim 1 , wherein the incubating is performed at a temperature of about 22° C. to about 37° C.
11. The method of claim 1 , wherein the composition further comprises a Tris buffer at a concentration sufficient to buffer in the basic pH range.
12. The method of claim 1 , wherein the composition further comprises sodium chloride.
13. The method of claim 1 , wherein the substrate is selected from the group consisting of cellulose membranes, porous polycarbonate, porous polytetrafluoroethylene, nylon membranes, glass filters, porous polyethyleneterephthalate, polymethylpentane, polypropylene, polyethylene and combinations thereof.
14. A method of preparing a cell-culturing system comprising:
(a) providing an extracellular matrix composition comprising an extracellular matrix; and at least one exogenous component selected from the group consisting of heparin, fibronectin and laminin;
(b) applying the composition to a substrate; and
(c) incubating the applied composition to allow polymerization thereof,
wherein the at least one exogenous component comprises heparin and fibronectin.
15. The method of claim 14 , further comprising laminin.