Method for processing tissues

Abstract

Methods for processing tissue are provided. In some embodiments, the methods comprise methods for decellularizing tissue samples by applying high hydrostatic pressure to the tissues samples. In some embodiments, the methods comprise methods for thawing tissue samples and/or reducing the bioburden in a sample by applying high hydrostatic pressure to the tissue samples.

1 . A method for decellularizing a tissue sample, comprising:
 providing a soft tissue sample comprising a mammalian tissue in a liquid; and   applying a pressure to the liquid of at least 200 MPa for a time sufficient to destroy substantially all of the native tissue cells within the soft tissue sample, wherein destroying substantially all of the cells includes disrupting the cell membrane of the cells such that washing the tissue sample in a saline solution allows removal of at least 95% of the native tissue cells.   
     
     
         2 . The method of  claim 1 , wherein the pressure applied to the liquid is at least about 500 MPa. 
     
     
         3 . The method of  claim 1 , wherein the pressure applied to the liquid is at least 300 MPa for at least 30 minutes. 
     
     
         4 . The method of  claim 1 , wherein the pressure applied to the liquid is at least 400 MPa for at least 10 minutes. 
     
     
         5 . The method of  claim 1 , wherein the pressure applied to the liquid is at least 400 MPa for at least 30 minutes. 
     
     
         6 . The method of  claim 1 , wherein the liquid comprises an aqueous salt solution. 
     
     
         7 . The method of  claim 6 , wherein the liquid comprises phosphate buffered saline. 
     
     
         8 . The method of  claim 1 , further comprising performing a sterilization process on the tissue. 
     
     
         9 . The method of  claim 8 , wherein the sterilization process comprises a gamma irradiation process. 
     
     
         10 . The method of  claim 8 , wherein the sterilization process comprises an e-beam irradiation process. 
     
     
         11 . The method of  claim 8 , wherein the sterilization process comprises a supercritical carbondioxide sterilization process. 
     
     
         12 . The method of  claim 8 , wherein the sterilization process comprises a peracetic acid treatment process. 
     
     
         13 . (canceled) 
     
     
         14 . The method of  claim 1 , wherein the mammalian tissue comprises skin. 
     
     
         15 . The method of  claim 1 , wherein the mammalian tissue comprises porcine dermis. 
     
     
         16 . The method of  claim 14 , further comprising removing an epidermal layer from the skin prior to applying the pressure. 
     
     
         17 . The method of  claim 1 , wherein the mammalian tissue comprises at least one of a tendon or ligament. 
     
     
         18 . The method of  claim 1 , wherein the mammalian tissue comprises small intestine submucosa. 
     
     
         19 . The method of  claim 1 , further comprising washing the soft tissue sample to remove destroyed cells; and
 performing at least one additional decellularization process on the soft tissue sample.   
     
     
         20 - 74 . (canceled) 
     
     
         75 . The method of  claim 1 , wherein the temperature of the tissue sample does not exceed 30° C. 
     
     
         76 . The method of  claim 75 , wherein the temperature of the tissue sample does not exceed 25° C.