Functionalization of plant tissues for human cell expansion

Abstract

Decellularized plant tissues and the use of these plant tissues as scaffolds are disclosed herein. Particularly, decellularized plant tissues are functionalized such to allow for human cell adhesion, thereby allowing for their use as scaffolds for human cells. These scaffolds can then be used in a number of applications/markets, including as research tools for tissue engineering, regenerative medicine, and basic cellular biology.

What is claimed is: 
     
         1 . A plant scaffold comprising a decellularized plant tissue comprising a plant adhesion molecule conjugated to a cell adhesion peptide. 
     
     
         2 . (canceled) 
     
     
         3 . The plant scaffold as set forth in  claim 1 , wherein the plant adhesion molecule is selected from the group consisting of a dopamine-containing compound, a polyphenol and combinations thereof. 
     
     
         4 . (canceled) 
     
     
         5 . The plant scaffold as set forth in  claim 1 , wherein the cell adhesion peptide is selected from the group consisting of RGD, RGDS (SEQ ID NO:7), CRGDS (SEQ ID NO:8), CRGDSP (SEQ ID NO:9), PHSRN (SEQ ID NO:10), GWGGRGDSP (SEQ ID NO:11), SIDQVEPYSSTAQ (SEQ ID NO:12), GRNIAEIIKDI (SEQ ID NO:13), DITYVRLKF (SEQ ID NO:14), DITVTLNRL (SEQ ID NO:15), GRYVVLPR (SEQ ID NO:16), GNRWHSIYITRFG (SEQ ID NO:17), GASIKVAVSADR (SEQ ID NO:18), GTTVKYIFR (SEQ ID NO:19), GSIKIRGTYS (SEQ ID NO:20), GSINNNR (SEQ ID NO:21), SDPGYIGSR (SEQ ID NO:22), YIGSR (SEQ ID NO:23), GTPGPQGIAGQGVV (SEQ ID NO:24), GTPGPQGIAGQRVV (SEQ ID NO:25), MNYYSNS (SEQ ID NO:26), KKQRFRHRNRKG (SEQ ID NO:27), CRGDGGGGGGGGGGGGGPHSRN (SEQ ID NO:28), CPHSRNSGSGSGSGSGRGD (SEQ ID NO:29), Acetylated-GCYGRGDSPG (SEQ ID NO:30), CRDGS (SEQ ID NO:31), cyclic RGD{Fd}C (SEQ ID NO:32), RKRLQVQLSIRT (SEQ ID NO:33), IKVAV (SEQ ID NO:34), YIGSR (SEQ ID NO:35), KRTGQYKL (SEQ ID NO:36), TYRSRKY (SEQ ID NO:37), KRTGQYKLGSKTGPGQK (SEQ ID NO:38), QAKHKQRKRLKSSC (SEQ ID NO:39), SPKHHSQRARKKKNKNC (SEQ ID NO:40), XBBXBX, wherein B=basic residue and X=hydropathic residue (SEQ ID NO:41), XBBBXXBX, wherein B=basic residue and X=hydropathic residue (SEQ ID NO:42), and RGDSP (SEQ ID NO:43). 
     
     
         6 . A pant scaffold comprising a decellularized plant tissue and a mineral layer. 
     
     
         7 . (canceled) 
     
     
         8 . The plant scaffold as set forth in  claim 6 , wherein the mineral layer comprises a calcium to phosphate ratio of from about 2.5:1 to about 1:1. 
     
     
         9 . The plant scaffold as set forth in  claim 6 , wherein the mineral layer further comprises a biomolecule selected from the group consisting of nucleic acids, proteins, peptides, growth factors, proteoglycans, and combinations thereof. 
     
     
         10 . (canceled) 
     
     
         11 . The plant scaffold as set forth in  claim 9 , wherein the biomolecule is a growth factor selected from the group consisting of 
     
     
         12 . A method for preparing a plant scaffold for tissue engineering, the method comprising:
 decellularizing a plant tissue; and   contacting the decellularized plant tissue with a plant adhesion molecule conjugated to a cell adhesion peptide.   
     
     
         13 . The method as set forth in  claim 12 , wherein the decellularizing the plant tissue comprises perfusing the plant tissue with at least one detergent. 
     
     
         14 . (canceled) 
     
     
         15 . The method as set forth in  claim 12 , wherein the plant adhesion molecule is selected from the group consisting of a dopamine-containing compound, a polyphenol and combinations thereof. 
     
     
         16 . (canceled) 
     
     
         17 . The method as set forth in  claim 12 , wherein the cell adhesion peptide is selected from the group consisting of RGD, RGDS (SEQ ID NO:7), CRGDS (SEQ ID NO:8), CRGDSP (SEQ ID NO:9), PHSRN (SEQ ID NO:10), GWGGRGDSP (SEQ ID NO:11), SIDQVEPYSSTAQ (SEQ ID NO:12), GRNIAEIIKDI (SEQ ID NO:13), DITYVRLKF (SEQ ID NO:14), DITVTLNRL (SEQ ID NO:15), GRYVVLPR (SEQ ID NO:16), GNRWHSIYITRFG (SEQ ID NO:17), GASIKVAVSADR (SEQ ID NO:18), GTTVKYIFR (SEQ ID NO:19), GSIKIRGTYS (SEQ ID NO:20), GSINNNR (SEQ ID NO:21), SDPGYIGSR (SEQ ID NO:22), YIGSR (SEQ ID NO:23), GTPGPQGIAGQGVV (SEQ ID NO:24), GTPGPQGIAGQRVV (SEQ ID NO:25), MNYYSNS (SEQ ID NO:26), KKQRFRHRNRKG (SEQ ID NO:27), CRGDGGGGGGGGGGGGGPHSRN (SEQ ID NO:28), CPHSRNSGSGSGSGSGRGD (SEQ ID NO:29), Acetylated-GCYGRGDSPG (SEQ ID NO:30), CRDGS (SEQ ID NO:31), cyclic RGD{Fd}C (SEQ ID NO:32), RKRLQVQLSIRT (SEQ ID NO:33), IKVAV (SEQ ID NO:34), YIGSR (SEQ ID NO:35), KRTGQYKL (SEQ ID NO:36), TYRSRKY (SEQ ID NO:37), KRTGQYKLGSKTGPGQK (SEQ ID NO:38), QAKHKQRKRLKSSC (SEQ ID NO:39), SPKHHSQRARKKKNKNC (SEQ ID NO:40), XBBXBX, wherein B=basic residue and X=hydropathic residue (SEQ ID NO:41), XBBBXXBX, wherein B=basic residue and X=hydropathic residue (SEQ ID NO:42), and RGDSP (SEQ ID NO:43). 
     
     
         18 . A method for preparing a plant scaffold for tissue engineering, the method comprising:
 decellularizing a plant tissue;   contacting the decellularized plant tissue with a modified simulated body fluid to form a mineral layer on the decellularized plant tissue, the modified simulated body fluid comprising at least one of calcium, phosphate, and carbonate.   
     
     
         19 . The method as set forth in  claim 18 , wherein the decellularizing the plant tissue comprises perfusing the plant tissue with at least one of a detergent and an enzyme. 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . The method as set forth in  claim 18 , wherein the contacting the decellularized plant tissue with a modified simulated body fluid comprises incubating the decellularized plant tissue in the modified simulated body fluid, wherein the modified simulated body fluid comprises from about 5 mM to about 12.5 mM calcium ions, from about 2 mM to about 12.5 mM phosphate ions, from about 4 mM to about 150 mM carbonate ions, and a pH of from about 4 to about 7.5. 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . The method as set forth in  claim 18 , wherein upon incubating the decellularized plant tissue with the modified simulated body fluid, a mineral layer is formed comprising a calcium to phosphate ratio of from about 2.5:1 to about 1:1. 
     
     
         26 . The method as set forth in  claim 18 , wherein the mineral layer further comprises a biomolecule selected from the group consisting of growth factors, proteoglycans, and combinations thereof. 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . A method of engineering mammalian tissue, the method comprising:
 decellularizing a plant tissue to provide a plant scaffold having a perfusable structure; and   culturing a cell within the plant scaffold.   
     
     
         30 . The method as set forth in  claim 29 , wherein the plant scaffold supports vasculature above about 200 μm diffusion limit. 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . A cell culture method comprising contacting a cell with the plant scaffold as set forth in  claim 1 . 
     
     
         34 . (canceled) 
     
     
         35 . A cell culture method comprising contacting a cell with the plant scaffold as set forth in  claim 6 . 
     
     
         36 . (canceled)