Thermolysin Treatment: A New Method for Dermo-epidermal Separation

• Published in: Journal of investigative dermatology Vol. 92; no. 1; pp. 78 - 81
• Main Authors: Walzer, Claude, Benathan, Messod, Frenk, Edgar
• Format: Journal Article
• Language: English
• Published: Danvers, MA Elsevier Inc 01.01.1989; Nature Publishing
• Subjects: Adult; Aged; Basement Membrane - drug effects; Biological and medical sciences; Child; Child, Preschool; Dermatology; Epidermis - drug effects; Female; Histological Techniques; Humans; Investigative techniques, diagnostic techniques (general aspects); Medical sciences; Microscopy, Electron; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Skin - anatomy & histology; Skin - drug effects; Skin - ultrastructure; Thermolysin - pharmacology; Trypsin - pharmacology; Pathology; Epidermis; Skin; Separation; Technique; Derm
• ISSN: 0022-202X; 1523-1747
• DOI: 10.1111/1523-1747.ep13071253

The epidermis of superficial human skin samples could easily be separated from the dermis following incubation at +4°C for 1h 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.