Method for processing and preserving collagen-based tissues for transplantation

Abstract

A method for processing and preserving an acellular collagen-based tissue matrix for transplantation is disclosed. The method includes the steps of processing biological tissues with a stabilizing solution to reduce procurement damage, treatment with a processing solution to remove cells, treatment with a cryoprotectant solution followed by freezing, drying, storage and rehydration under conditions that preclude functionally significant damage and reconstitution with viable cells.

1 . A method of making a tissue matrix, the method comprising: 
 (a) procuring a collagen-based tissue; and    (b) incubating the collagen-based tissue in a processing solution to produce processed tissue, wherein the processing solution extracts viable cells from the structural protein and collagen matrix of the collagen-based tissue,    wherein the processed tissue comprises a basement membrane to which viable endothelial cells or viable epithelial cells securely attach.    
     
     
         2 . The method of  claim 1 , further comprising the steps of: 
 (c) cryopreparing the processed tissue by incubation in a cryoprotective solution; and    (d) freezing the processed tissue to produce a frozen, processed tissue,    wherein the frozen, processed tissue comprises a basement membrane to which, after thawing of the frozen, processed tissue, viable endothelial cells or viable epithelial cells securely attach.    
     
     
         3 . The method of  claim 2 , further comprising the step of: 
 (e) drying the frozen, processed tissue to produce a dried, processed tissue,    wherein the dried, processed tissue comprises a basement membrane to which, after rehydration of the dried, processed tissue, viable endothelial cells or viable epithelial cells securely attach.    
     
     
         4 . The method of  claim 3 , further comprising the step of: 
 (f) incubating the dried, processed tissue in a rehydration solution to give a rehydrated tissue,    wherein the rehydrated tissue comprises a basement membrane to which viable endothelial cells or viable epithelial cells securely attach.    
     
     
         5 . The method of  claim 4 , further comprising the step of: 
 (g) inoculating the rehydrated tissue with autogeneic cells, allogeneic cells, or a combination of autogeneic cells and allogeneic cells,    wherein the rehydrated tissue, after the inoculation, comprises a basement membrane to which viable endothelial cells or viable epithelial cells securely attach.    
     
     
         6 . The method of  claim 1 , wherein the collagen-based tissue comprises dermis.  
     
     
         7 . The method of  claim 1 , wherein the collagen-based tissue comprises a tissue of venous or arterial origin.  
     
     
         8 . The method of  claim 7 , wherein the tissue of venous origin is saphenous vein tissue.  
     
     
         9 . The method of  claim 1 , wherein the collagen-based tissue comprises a heart valve.  
     
     
         10 . The method of  claim 1 , wherein the collagen-based tissue comprises heart valve leaflet tissue.  
     
     
         11 . The method of  claim 1 , wherein the collagen-based tissue is a human collagen-based tissue.  
     
     
         12 . The method of  claim 1 , wherein the collagen-based tissue is a non-human animal collagen-based tissue.  
     
     
         13 . The method of  claim 12 , wherein the non-human animal is a pig.  
     
     
         14 . The method of  claim 12 , wherein the non-human animal is a dog.  
     
     
         15 . The method of  claim 1 , wherein the collagen-based tissue comprises a tissue selected from the group consisting of ligament tissue, tendon tissue, bone tissue, cartilage tissue, dura mater tissue, and nerve tissue.  
     
     
         16 . A tissue matrix made by the method of  claim 1 .  
     
     
         17 . A method of treatment comprising grafting the tissue matrix of  claim 16  to or into a patient.  
     
     
         18 . The method of  claim 17 , wherein the patient is selected from the group consisting of a burn patient, a diabetic patient, a patient with pressure ulcers, a patient requiring reconstructive surgery, and a patient requiring skin replacement following excision of a skin lesion.