Method for processing tissues
View Patent ↗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 reducing the bioburden in a soft tissue sample, comprising:
providing a soft tissue sample comprising a mammalian skin containing intact dermal cells and having at least a portion of non-dermal components removed in a container comprising an aqueous solution; and
applying a pressure to the aqueous solution and time sufficient to cause at least a 5 log reduction in a bacterial load of the skin sample and to destroy substantially all of the cells within the skin sample, wherein destroying substantially all of the cells includes disrupting the cell membrane of the cells such that washing the soft tissue sample in a saline solution allows removal of at least 95% of the native skin cells, and wherein applying the pressure includes increasing the pressure at a rate to control a temperature of the skin sample such that the temperature of the skin sample does not exceed 30° C.
2. The method of claim 1 , wherein the pressure applied to the aqueous solution is at least 500 MPa.
3. The method of claim 1 , wherein the pressure applied to the aqueous solution is at least 300 MPa for at least 30 minutes.
4. The method of claim 1 , wherein the pressure applied to the aqueous solution is at least 400 MPa for at least 10 minutes.
5. The method of claim 1 , wherein the pressure applied to the aqueous solution is at least 400 MPa for at least 40 minutes.
6. The method of claim 1 , wherein the aqueous solution comprises an aqueous salt solution.
7. The method of claim 6 , wherein the aqueous solution comprises a 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 comprise a supercritical carbon dioxide sterilization process.
12. The method of claim 8 , wherein the sterilization process comprises a peracetic acid treatment process.
13. The method of claim 1 , wherein the mammalian tissue comprises porcine dermis.
14. The method of claim 13 , further comprising removing an epidermal layer from the skin prior to applying the pressure.