Decellularization of Plant Cell Culture Materials for Tissue Engineering and Drug Delivery

Abstract

Provided herein are enzymatically decellularized cells, and methods of producing said cells, that can be used in a scaffold. The scaffolds featured herein are biocompatible and can comprise decellularized cells that have been modified to express a bioactive agent or molecule.

1 . A method for decellularizing cells, the method comprising:
 contacting a plurality of cells with a composition comprising a nuclease, thereby decellularizing the plurality of cells, wherein the plurality of cells are cellulose producing cells.   
     
     
         2 . The method of  claim 1 , wherein the nuclease is DNasel. 
     
     
         3 . The method of  claim 1 , wherein the plurality of cells are plant cells. 
     
     
         4 . The method of  claim 1  further comprising culturing the plurality of cells prior to contacting the cells with the nuclease. 
     
     
         5 . The method of  claim 1 , further comprising isolating cellular material from the cultured plurality of cells. 
     
     
         6 . A scaffold comprising decellularized cells derived from a cellulose producing organism and a bioactive molecule. 
     
     
         7 . The scaffold of  claim 6 , wherein the bioactive molecule is attached to the decellularized cells. 
     
     
         8 . The scaffold of  claim 6 , wherein the decellularized cells are enzymatically decellularized plant-derived cells. 
     
     
         9 . The scaffold of  claim 6 , wherein the bioactive molecule is VEGF, bFGF, IL-2, or a molecule that directs mammalian cell expansion, differentiation, or a cellular response. 
     
     
         10 . The scaffold of  claim 6 , wherein the decellularized cells are cultured cells. 
     
     
         11 .- 13 . (canceled) 
     
     
         14 . A method of producing a biocompatible scaffold, the method comprising:
 contacting the cellulose producing cells with a composition comprising a nuclease, thereby producing decellularized cells; and   creating a scaffold from the decellularized modified cells.   
     
     
         15 . The method of  claim 14  further comprising modifying the cellulose producing cells to express a bioactive agent. 
     
     
         16 . The method of  claim 14 , wherein the bioactive agent is a molecule that directs mammalian cell expansion, differentiation, or a cellular response. 
     
     
         17 . The method of  claim 14 , wherein the bioactive agent is imbedded, attached, or associated with the cell walls of the cellulose producing cells. 
     
     
         18 . The method of  claim 14 , wherein the plurality of cellulose producing cells are cultured cells. 
     
     
         19 . The method of  claim 14 , wherein the nuclease is DNasel. 
     
     
         20 . The method of  claim 14 , wherein the composition further comprises trypsin, collagenase, lipase, dispase, thermolysin, and α-galactosidase.