Tissue scaffolds derived from forestomach extracellular matrix

Abstract

The present invention pertains to the development of Extracellular Matrix (ECM) scaffolds derived from the forestomach of a ruminant. Such scaffolds are useful in many clinical and therapeutic applications, including wound repair, tissue regeneration, and breast reconstruction. In addition, the present invention features methods of isolating ECM scaffolds from mammalian organs, including but not limited to the ruminant forestomach. The invention further features laminated ECM scaffolds containing a polymer positioned between individual ECM sheets. The polymer may optionally contain bioactive molecules to enhance the functionality of the scaffold.

What is claimed: 
     
         1 . A method of separating or decellularising the layers within all or a portion of a tissue, comprising creating a transmural osmotic flow between two sides of the tissue, such that the layers within all or a portion of the tissue are separated or decellularised. 
     
     
         2 . The method of  claim 1 , wherein transmural osmotic flow is created from the luminal to the abluminal side of the tissue. 
     
     
         3 . The method of  claim 1 , comprising removing all or part of a tissue layer selected from the group consisting of epithelium, basement membrane, tunica muscularis, and combinations thereof. 
     
     
         4 . The method of  claim 1 , wherein the tissue comprises a keratinized stratified squamous epithelium. 
     
     
         5 . The method of  claim 1 , further comprising the step of encapsulating a first solution within the tissue or portion thereof, and immersing the tissue or portion thereof in a second solution which is hypertonic to the first solution. 
     
     
         6 . The method of  claim 5 , further comprising the step of removing the tissue or portion thereof from the second solution, and immersing the tissue or portion thereof in a third solution which is also hypertonic to the first solution. 
     
     
         7 . The method of  claim 1 , further comprising the step of encapsulating a first solution within the tissue or portion thereof, and immersing the tissue or portion thereof in a second solution which is hypotonic to the first solution. 
     
     
         8 . The method of  claim 7 , further comprising the step of removing the tissue or portion thereof from the second solution, and immersing the tissue or portion thereof in a third solution which is also hypotonic to the first solution. 
     
     
         9 . The method of  claim 7 , wherein the second solution comprises water, optionally including at least one buffer, detergent or salt. 
     
     
         10 . The method of  claim 7 , wherein the first solution comprises 4 M NaCl. 
     
     
         11 . The method of  claim 8 , wherein the first solution comprises 4 M NaCl, the second solution comprises 0.028% Triton X-200 and 0.1% EDTA, and the third solution comprises 0.028% SDS. 
     
     
         12 . The method of  claim 8 , wherein the first solution comprises 4 M NaCl, the second solution comprises 0.1% SDS, and the third solution comprises 0.028% Triton X-200 and 0.1% EDTA. 
     
     
         13 . The method of  claim 1 , wherein the method is performed at a temperature between 2° C. and 4° C. in less than 36 hours. 
     
     
         14 . The method of  claim 1 , wherein the method is performed at 4° C. 
     
     
         15 . The method of  claim 1 , wherein the method is performed in less than 24 hours. 
     
     
         16 . The method of  claim 10 , wherein the second solution comprises 0.028% Triton X-200, 0.1% EDTA, and 0.1% SDS. 
     
     
         17 . The method of  claim 16 , wherein the method is performed at a temperature between 18° C. and 24° C. 
     
     
         18 . The method of  claim 16 , wherein the method is performed in less than 6 hours. 
     
     
         19 . The method of  claim 1 , wherein the tissue or portion thereof is distended to increase the transmural osmotic flow. 
     
     
         20 . The method of  claim 1 , wherein the tissue is a whole organ, or a portion thereof. 
     
     
         21 . The method of  claim 1 , wherein the tissue is derived from the forestomach of a ruminant. 
     
     
         22 . The method of  claim 21 , wherein the tissue is the rumen. 
     
     
         23 . The method of  claim 21 , wherein the ruminant belongs to a genus selected from the group consisting of  Capra, Bos, Cervus  and  Ovis.   
     
     
         24 . The method of  claim 23 , wherein the ruminant is  Ovis aries.   
     
     
         25 . A tissue scaffold produced according to the method of  claim 1 . 
     
     
         26 . A method of forming a tissue scaffold, comprising performing the method of  claim 1 .