Thermolysin in human cultured keratinocyte isolation

Brazilian Journal of Biology

A. Gragnani¹  Cs. Sobral¹  Lm. Ferreira¹ 

[1] ,Federal University of São Paulo Surgery Department Division of Plastic SurgerySão Paulo SP ,Brazil

关键词: keratinocyte;    thermolysin;    trypsin;    colony forming efficiency;    queratinócitos;    termolisina;    tripsina;    formação de colônias;   

DOI  :  10.1590/S1519-69842007000100014

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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