Novel peptides for supporting endothelial progenitor cell rolling and capture and endothelialization of biomaterials

Abstract

The present invention relates to the production of endothelialized matrices and materials from immature endothelial cells using substrates to which particular peptides have been grafted. The resultant substrates can be used to capture and support immature endothelial cells. Further, the methods and compositions of the present invention provide viable cell delivery platforms that allow for production and provision of endothelialized medical devices and implants, including vascular grafts, stents, shunts, and valves, endothelialized surfaces and channels for in vitro testing devices, including microfluidic chips, and materials that support vascularization such as for use in engineered tissues. The present invention includes novel methods required for the successful production of cellularized substrates, systems and components used for the same, and methods of using the resultant cell and tissue compositions.

1 - 19 . (canceled) 
     
     
         20 . A method of capturing endothelial cells from a flowing fluid, the method comprising:
 providing a substrate that is operably linked to at least one first peptide comprising SEQ ID NO: 1 and at least one second peptide comprising SEQ ID NO: 2; and   exposing the substrate to a source of endothelial cells in a flowing fluid.   
     
     
         21 . The method of  claim 20 , wherein the endothelial cells are immature endothelial cells, endothelial progenitor cells, or stem cell-derived endothelial cells. 
     
     
         22 . The method of  claim 20 , wherein the substrate is further operably linked to at least one additional peptide selected from the group consisting of SEQ ID NOs: 8, 11 and 18 and combinations thereof. 
     
     
         23 . The method of  claim 20 , wherein providing a substrate comprises providing a medical device or implant. 
     
     
         24 . The method of  claim 23 , wherein providing a medical device or implant comprises providing a vascular graft, stent, shunt, or valve. 
     
     
         25 . The method of  claim 20 , wherein providing a substrate comprises providing an in vitro testing device. 
     
     
         26 . The method of  claim 20 , wherein the substrate comprises a hydrogel. 
     
     
         27 . The method of  claim 20 , wherein the substrate comprises a polymer. 
     
     
         28 . The method of  claim 20 , wherein the substrate comprises biomimetic material. 
     
     
         29 . The method of  claim 28 , wherein the substrate comprises decellularized extracellular matrix (dECM). 
     
     
         30 . The method of  claim 20 , wherein the molar ratio of the second peptide to the first peptide is between 3:1 and 1:3. 
     
     
         31 . The method of  claim 20 , wherein the flowing fluid is blood. 
     
     
         32 . A composition for capturing endothelial cells from a flowing fluid, the composition comprising:
 a substrate;   at least one first peptide comprising SEQ ID NO: 1 operatively linked to the surface of the substrate; and   at least one second peptide comprising SEQ ID NO: 2 operatively linked to the surface of the substrate;   wherein the composition facilitates the capture of endothelial cells from a flowing fluid.   
     
     
         33 . The composition of  claim 32 , further comprising at least one additional peptide selected from the group consisting of SEQ ID NOs: 8, 11 and 18 and combinations thereof. 
     
     
         34 . The composition of  claim 32 , wherein the endothelial cells are immature endothelial cells, endothelial progenitor cells, or pluripotent stem cell-derived endothelial cells. 
     
     
         35 . The composition of  claim 32 , wherein the substrate comprises a polymer. 
     
     
         36 . The composition of  claim 32 , wherein said substrate comprises a hydrogel. 
     
     
         37 . The composition of  claim 32 , wherein said substrate comprises a biomimetic material. 
     
     
         38 . The composition of  claim 37 , wherein the substrate comprises decellularized extracellular matrix (dECM). 
     
     
         39 . The composition of  claim 32 , wherein the molar ratio of the second peptide to the first peptide is between 3:1 and 1:3. 
     
     
         40 . A composition for capturing endothelial cells from a flowing fluid, the composition comprising:
 a substrate;   at least one first peptide comprising SEQ ID NO: 8 operatively linked to the surface of the substrate; and   at least one second peptide comprising SEQ ID NO: 2 operatively linked to the surface of the substrate;   wherein the composition facilitates the capture of endothelial cells from a flowing fluid.   
     
     
         41 . A method of capturing endothelial cells from a flowing fluid, the method comprising:
 providing the composition of  claim 40 ; and   exposing the substrate to a source of endothelial cells in a flowing fluid.