Thermolysin in human cultured keratinocyte isolation

• Autor(a) principal: Gragnani,A.
• Data de Publicação: 2007
• Outros Autores: Sobral,CS., Ferreira,LM.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Brazilian Journal of Biology
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1519-69842007000100014
• topic: keratinocyte; thermolysin; trypsin; colony forming efficiency

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.