Cryopreservation of viable human skin substitutes
AssigneeStratatech Corporation
InventorsB. Lynn Allen-Hoffmann, John C. Pirnstill, Kenneth R. Gratz, Allen R. Comer
Life of this patent
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Abstract
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 cryopreserving viable skin substitutes.
Claims29
A method of cryopreserving an organotypically cultured skin equivalent to maintain viable tissue comprising:
- treating an organotypically cultured skin equivalent in a cryoprotectant solution in a single step;
- packaging said organotypically cultured skin equivalent to provide a packaged skin equivalent;
- and
- freezing said packaged organotypically cultured skin equivalent to provide a cryopreserved skin equivalent.
A method of thawing a cryopreserved skin equivalent prior to application to a patient, comprising:
- warming said cryopreserved skin equivalent;
- and
- contacting said cryopreserved skin equivalent with a diffusion mediator comprising a tissue compatible solution to allow removal of said cryoprotectant solution by diffusion.
A cryopreserved skin equivalent equilibrated with a cryoprotectant, said skin equivalent being substantially free of excess cryoprotectant on the exterior surface of said skin equivalent.
A kit comprising a cryopreserved skin substitute, a diffusion mediator, and a tissue compatible solution.
A method comprising:
- providing a culture dish comprising a cell culture substrate movable between defined upper and lower positions in said culture dish,
- forming a skin equivalent on said cell culture substrate, wherein said cell culture substrate is at said upper position,
- lowering said cell culture substrate to said lower position for further processing.
A method of producing a cryopreserved skin equivalent comprising:
- providing a culture dish comprising an insert movable between upper and lower positions in said culture dish, said insert having a bottom planar surface formed from a porous membrane,
- forming a dermal equivalent comprising fibroblast cells on said porous membrane in said insert, wherein said insert is placed in said upper position said in culture dish,
- culturing said fibroblast cells to form a dermal equivalent,
- applying keratinocytes to said dermal equivalent,
- culturing said keratinocytes in a culture medium under conditions such that said keratinocytes form a skin equivalent comprising stratified epithelium,
- removing said culture medium,
- lowering said insert to said lower position,
- treating said skin equivalent with a cryoprotectant solution, and
- freezing said skin equivalent in said culture dish.
Dependencies are inferred from the claim text.