Thermolysin in human cultured keratinocyte isolation.
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.


OBJECTIVE
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.


CONCLUSIONS
The number of colonies in the thermolysin group was significantly greater than in the trypsin group.
Gragnani
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Division of Plastic Surgery, Surgery Department, Federal University of São Paulo - UNIFESP/EPM, São Paulo, SP, Brazil. alfredogf@ig.com.br
Sobral
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Ferreira
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D002469:Cell Separation / Q000379:methods*; D003114:Colony-Forming Units Assay; D006801:Humans; D007231:Infant, Newborn; D015603:Keratinocytes / Q000166:cytology* / Q000637:transplantation; D013820:Thermolysin / Q000494:pharmacology*; D023822:Tissue Engineering; D014357:Trypsin / Q000494:pharmacology*
D014357:Trypsin; D013820:Thermolysin