Keratinocyte culture and uses thereof
AssigneeEpitech Corporation, Ltd.
InventorsThomas Hunziker, Alain Limat
Life of this patent
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Abstract
The present invention relates to the treatment of skin defects by organotypically-cultured autologous keratinocytes isolated from the outer root sheath of anagenic or growing hair. Methods for primary, as well as subsequent organotypic cultures (i.e., epidermal equivalents) in fully-defined media supplemented by autologous human serum and substances isolated from blood components, with minimal allogeneic biological supplements, are disclosed herein. Techniques to prepare epidermal equivalents for transplantation by use of a biocompatible glue are also disclosed herein.
Claims22
A method for the treatment of a skin defect comprising
- (a) culturing an intact hair follicle of an anagenic hair to obtain outer root sheath cells;
- (b) culturing said outer root sheath cells in a medium containing human serum in a concentration of less than 5% to obtain keratinocyte precursor cells;
- (c) preparing an epidermnal or dermal equivalent comprising said keratinocyte precursor cells, wherein said keratinocyte precursor cells are seeded at a density of between 3×104 cells/cm2 and 1×105 cells/cm2;
- and
- (d) applying a portion of said epidermal or dermal equivalent to said defect.
A method for the treatment of a skin defect comprising
- (a) culturing an intact hair follicle of an anagenic hair to obtain outer root sheath cells;
- (b) culturing said outer root sheath cells to obtain keratinocyte precursor cells;
- (c) preparing an epidermal or dermal equivalent comprising said keratinocyte precursor cells wherein said keratinocyte precursor cells are seeded at a density of between 3×104 cells/cm2 and 1×105 cells/cm2;
- and
- (d) applying a portion of said epidermal or dermal equivalent to said defect wherein all culturing of cells is performed in a medium which utilizes autologous or homologous human serum in a concentration of preferably less than approximately 5%.
A method for the treatment of a skin defect comprising
- (a) culturing an intact hair follicle of an anagenic hair to obtain outer root sheath cells;
- (b) culturing said outer root sheath cells in a medium containing human serum in a concentration of less than 5% to obtain keratinocyte precursor cells;
- (c) preparing an epidermal or dermal equivalent comprising said keratinocyte precursor cells wherein said keratinocyte precursor cells are seeded at a density of between 3×104 cells/cm2 and 1×105 cells/cm2;
- and
- (d) applying a portion of said epidermal or dermal equivalent to said defect wherein said epidermal or dermal equivalent is coated on its top or cornified side with a fibrin glue.
Dependencies are inferred from the claim text.