Patent ID: 6730513
Filing Date: 2004-05-04
Classification: A61L,A61P

Abstract:
A method for the selection of keratinocyte precursor cells from the outer root sheath of hair for subsequent use in a composition for healing a skin defect, comprising the step of:(a) plucking of an anagen or growing hair; (b) primary-culturing the outer root sheath-derived keratinocyte precursor cells by adhering said anagen hair, in toto, to a microporous membrane, which possesses growth-arrested limited feeder cells on its undersurface so as to select for keratinocyte precursor cells from the outer root sheath of hair, wherein the primary culture medium contains human serum in a concentration less than 5%; (c) organotypically-culturing the outer root sheath cells harvested from said primary cultures by modulating a microporous membrane which also possesses growth-arrested limited feeder cells on its undersurface, wherein the organotypic culture medium contains human serum in a concentration less than 5% and the keratinocyte precursor cells are seeded at a density of between 3Ã—104 cells/cm2 and 1Ã—105 cells/cm2; (d) generating an epidermal or complex skin equivalent, for subsequent use as a graft insert, comprised of keratinocyte precursor cells by placing a carrier membrane on top of said organotypic-culture from step (c) and detaching said skin or epidermal equivalent, which is comprised of the keratinocyte precursor cells and carrier membrane, together as a single, laminat unit; (e) contacting said epidermal or skin equivalent with a skin defect present on an individual, and immobilizing said epidermal or skin equivalent at the site of contact.