Thermolysin in human cultured keratinocyte isolation

• Autor(a) principal: Gragnani, Alfredo [UNIFESP]
• Data de Publicação: 2007
• Outros Autores: Sobral, Christiane Steponavicius [UNIFESP], Ferreira, Lydia Masako [UNIFESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: http://dx.doi.org/10.1590/S1519-69842007000100014; http://repositorio.unifesp.br/handle/11600/3516
• topic: keratinocyte; thermolysin; trypsin; colony forming efficiency; queratinócitos; termolisina; tripsina; formação de colônias

BACKGROUND: When treating extensively burned patients using cultured epidermal sheets, the main problem is the time required for its production. Conventional keratinocyte isolation is usually done using Trypsin. We used a modification of the conventional isolation method in order to improve this process and increase the number of colonies from the isolated epidermal cell population. PURPOSE: To compare the action of trypsin and thermolysin in the keratinocyte isolation using newborn foreskin. METHODS: This method used thermolysin as it selectively digests the dermo-epidermal junction. After dermis separation, the epidermis was digested by trypsin in order to obtain a cell suspension. RESULTS: Compared to the conventional procedure, these experiments demonstrated that in the thermolysin group, the epidermis was easily detached from the dermis, there was no fibroblast contamination and there were a larger number of keratinocyte colonies which had a significant statistical difference. CONCLUSION: The number of colonies in the thermolysin group was significantly greater than in the trypsin group.