Dried and irradiated skin equivalents for ready use
View Patent ↗The present invention relates generally to systems and methods for preparing, storing, shipping and using skin equivalents made by organotypic culture. In particular, the present invention relates to systems and methods for producing, transporting, storing and using skin equivalents produced by organotypic culture at reduced temperatures, preferably from 2-8 degrees Celsius to ambient temperature. The methods include sterile packaging of the grafts so that the sterility and integrity of the package is maintained until the time of use for grafting purposes.
1. A method of preserving an organotypically cultured skin equivalent for use as a wound dressing comprising:
providing said organotypically cultured skin equivalent and a package,
wherein the organotypically cultured skin equivalent comprises keratinocytes that have stratified into squamous epithelia;
treating said skin equivalent to eliminate cell viability in the skin equivalent; and
packaging said skin equivalent to provide a packaged skin equivalent.
2. The method of claim 1 , wherein said treating step comprises irradiating said skin equivalent so that said skin equivalent is rendered sterile and non-viable.
3. The method of claim 2 , wherein said irradiating is performed with gamma irradiation.
4. The method of claim 2 , wherein said irradiating occurs after packaging.
5. The method of claim 1 , wherein said treating step comprises drying said skin equivalent under conditions such that cells in said skin equivalent are rendered non-viable.
6. The method of claim 5 , wherein said drying is performed by a method selected from the group consisting of vacuum drying and freeze drying.
7. The method of claim 5 , wherein said skin equivalent is dried to a final mass of less than 50% of a wet skin equivalent.
8. The method of claim 5 , wherein said skin equivalent, after rehydration, has an initial DPM value of from 20 to 300, and a DPM change value of from 5 to 400.
9. The composition of claim 5 , wherein said skin equivalent, after rehydration, has a tensile strength of from 0.1 to 5.0 MPa.
10. The method of claim 1 , wherein said treating occurs before packaging.
11. The method of claim 1 , wherein said treating comprises drying said skin equivalent under conditions such that cells making up said skin equivalent are rendered non-viable and irradiating said skin equivalent under conditions such said skin equivalent is rendered sterile.
12. The method of claim 11 , wherein said drying step occurs before said packaging and said irradiation step occurs after said packaging step.
13. The method of claim 1 , wherein said organotypically cultured skin equivalent comprises NIKS cells.
14. The method of claim 13 , wherein said NIKS cells comprise an exogenous nucleic acid sequence encoding an antimicrobial polypeptide.
15. The method of claim 14 , wherein said antimicrobial polypeptide is selected from the group consisting of human beta-defensin 1, human beta-defensin 2, human beta-defensin 3, and cathelicidin.
16. The method of claim 14 , wherein said antimicrobial polypeptide is provided in a quantity of from 10 to 500 ng of antimicrobial polypeptide per milliliter of a surface extraction solution.
17. The method of claim 1 , wherein said package is heat sealable.