Abstract
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.
Highlights
In the last few decades, there have been major developments in skin cell culture (Prunieras et al, 1976) making its experimental (Rheinwald and Green, 1977; Rheinwald, 1980; Pasch et al, 1999) and clinical use (Gallico et al, 1984; Cuono et al, 1987; Boyce et al, 1993; Tompkins and Burke, 1996) possible
Skin from the same donor was used in each experiment, comparing the trypsin and thermolysin group
An easy separation at the dermal-epidermal junction was obtained; the whole epidermis with a complete basal cell layer was disaggregated with 2 sequential trypsin digestions of 30 minutes each
Summary
In the last few decades, there have been major developments in skin cell culture (Prunieras et al, 1976) making its experimental (Rheinwald and Green, 1977; Rheinwald, 1980; Pasch et al, 1999) and clinical use (Gallico et al, 1984; Cuono et al, 1987; Boyce et al, 1993; Tompkins and Burke, 1996) possible The disadvantages of this technique are still related to its high costs and time required for keratinocyte culture (Morgan and Yarmush, 1997). Conclusion: The number of colonies in the thermolysin group was significantly greater than in the trypsin group
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