Thermolysin in human cultured keratinocyte isolation

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 inc...

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Bibliographic Details
Published in:Brazilian journal of biology 2007-02, Vol.67 (1), p.105-109
Main Authors: Gragnani, A, Sobral, C S, Ferreira, L M
Format: Article
Language:English
Subjects:
BIOLOGY
Cell Separation - methods
colony forming efficiency
Colony-Forming Units Assay
Humans
Infant, Newborn
keratinocyte
Keratinocytes - cytology
Keratinocytes - transplantation
thermolysin
Thermolysin - pharmacology
Tissue Engineering
trypsin
Trypsin - pharmacology
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Summary: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. To compare the action of trypsin and thermolysin in the keratinocyte isolation using newborn foreskin. 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. 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. The number of colonies in the thermolysin group was significantly greater than in the trypsin group.
ISSN:1519-6984
1678-4375
1519-6984
DOI:10.1590/S1519-69842007000100014