Thermolysin treatment: a new method for dermo-epidermal separation

Abstract The epidermis of superficial human skin samples could easily be separated from the dermis following incubation at + 4°C for 1 h in a solution containing 250–500 μg/ml thermolysin, a proteolytic enzyme hitherto mostly used for protein analysis. Light and electron microscopy revealed that the dermo-epidermal separation occurred at the basement membrane between the sites of bullous pemphigoid antigen and laminin and that the hemidesmosomes were selectively disrupted. The cohesion and morphology of the separated epidermis as well as the immunologic parameters investigated were not altered by this procedure. The clear cut dermo-epidermal separation produced by thermolysin treatment differed from the separation obtained with trypsin, which predominantly occurred between basal and suprabasal cells by disruption of desmosomes.