Decellularization and recellularization of whole organs

Abstract

The present invention provides systems and methods for the decellularization and recellularization of tissue segments. In certain instances the invention comprises coating or sealing decellularized tissue segments with a cross-linked alginate hydrogel. The present invention also provides a decellularization kit, which may be used to generate decellularized tissue segments for high throughput studies. Also included are compositions and methods of tissue sealants comprising methacrylated alginate.

1 .- 64 . (canceled) 
     
     
         65 . A method for sealing a wound in a mammal comprising applying a sealing agent comprising methacrylated alginate and a crosslinking agent to the wound, wherein the crosslinking agent is selected from the group consisting of eosin Y, triethanolamine, calcium chloride; ammonium persulfate (APS) and tetramethylethylenediamine (TEMED), glutaraldehyde, an epoxide, oxidized dextran, p-azidobenzoyl hydrazide, N-[α.-maleimidoacetoxy]succinimide ester, p-azidophenyl glyoxal monohydrate, bis-[β-(4-azidosalicylamido)ethyl]disulfide, bis[sulfosuccinimidyl]suberate, dithiobis[succinimidyl proprionate, disuccinimidyl suberate, 1-ethyl-3[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), riboflavin, visible light irradiation, blue light irradiation, ultraviolet irradiation, and any combination thereof. 
     
     
         66 . The method of  claim 65 , wherein the methacrylated alginate is formulated in a liquid or a solid. 
     
     
         67 . The method of  claim 66 , wherein the solid is hydrated after application to the wound. 
     
     
         68 . The method of  claim 66 , wherein the application further comprises crosslinking the methacrylated alginate to form a hydrogel. 
     
     
         69 . The method of  claim 65 , wherein the application of methacrylated alginate comprises injecting the methacrylated alginate through a catheter. 
     
     
         70 . The method of  claim 69 , wherein the catheter is a fiber optic catheter capable of pulsing the methacrylated alginate with visible light. 
     
     
         71 . The method of  claim 65 , wherein the methacrylated alginate sealing agent further comprises at least one therapeutic agent or biological agent. 
     
     
         72 . A tissue sealant or coating for a biological tissue scaffold comprising methacrylated alginate and a crosslinking agent, wherein the crosslinking agent is selected from the group consisting of eosin Y, triethanolamine, calcium chloride; ammonium persulfate (APS) and tetramethylethylenediamine (TEMED), glutaraldehyde, an epoxide, oxidized dextran, p-azidobenzoyl hydrazide, N-[α.-maleimidoacetoxy]succinimide ester, p-azidophenyl glyoxal monohydrate, bis-[β-(4-azidosalicylamido)ethyl]disulfide, bis[sulfosuccinimidyl]suberate, dithiobis[succinimidyl proprionate, disuccinimidyl suberate, 1-ethyl-3[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), riboflavin, visible light irradiation, blue light irradiation, ultraviolet irradiation, and any combination thereof. 
     
     
         73 . The tissue sealant or coating of  claim 72 , wherein the methacrylated alginate is formulated in a liquid or a solid. 
     
     
         74 . The tissue sealant or coating of  claim 73 , wherein the solid is hydrated after application. 
     
     
         75 . The tissue sealant or coating of  claim 72 , wherein the methacrylated alginate sealing agent further comprises at least one therapeutic agent or biological agent. 
     
     
         76 . The tissue sealant or coating of  claim 75 , wherein the therapeutic agent or biological agent is selected from the list consisting of a nucleic acid, an antibiotic, an anti-inflammatory agent, a growth factor, a cytokine, an enzyme, a protein, a peptide, at least one amino acid, a carbohydrate, a lipid, a hormone, a microsome, one or more cells, a derivative or a variation thereof, and any combination thereof. 
     
     
         77 . A kit for recellularizing a decellularized biological tissue, comprising:
 at least one reagent for decellularizing a biological tissue;   the tissue sealant of  claim 72 ; and   instructions for decellularizing a biological tissue, applying the at least one sealing agent to at least a portion of the decellularized biological tissue, and contacting the decellularized biological tissue with at least one cell type.   
     
     
         78 . The kit of  claim 77 , further comprising an assay for detecting the presence of the at least one reagent on the decellularized tissue. 
     
     
         79 . A system for coating a tissue scaffold, comprising:
 a tissue scaffold;   the coating of claim  27 ; and   instructions for coating the tissue scaffold, applying the coating to at least a portion of the tissue scaffold, and contacting the tissue scaffold with at least one cell type.   
     
     
         80 . The system of  claim 79 , wherein the tissue scaffold is a decellularized biological tissue.