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Separation of Rat Epidermis and Dermis with Thermolysin to Detect Site-Specific Inflammatory mRNA and Protein
Published on: September 29, 2021
Thermolysin in human cultured keratinocyte isolation
A Gragnani1, C S Sobral, L M Ferreira
1Division of Plastic Surgery, Surgery Department, Federal University of São Paulo - UNIFESP/EPM, São Paulo, SP, Brazil. alfredogf@ig.com.br
Summary
Thermolysin enzyme treatment significantly enhances keratinocyte isolation for cultured epidermal sheets, yielding more colonies than conventional trypsin methods. This improved cell isolation is crucial for treating extensively burned patients.
Area of Science:
- Regenerative Medicine
- Tissue Engineering
- Dermatology
Background:
- Cultured epidermal sheets are vital for treating extensive burns.
- Current keratinocyte isolation methods using trypsin are time-consuming.
- Improving keratinocyte yield and colony formation is essential for efficient treatment.
Purpose of the Study:
- To compare the efficacy of thermolysin and trypsin in keratinocyte isolation.
- To evaluate the impact of thermolysin on dermo-epidermal junction digestion.
- To assess the resulting keratinocyte colony count and purity.
Main Methods:
- Utilized thermolysin for selective digestion of the dermo-epidermal junction.
- Separated dermis from epidermis using thermolysin.
- Employed trypsin for subsequent epidermal cell suspension digestion.
- Compared outcomes with conventional trypsin-only isolation.
Main Results:
- Thermolysin facilitated easier epidermis detachment from the dermis.
- Absence of fibroblast contamination was observed in the thermolysin group.
- Significantly higher numbers of keratinocyte colonies were obtained with thermolysin.
Conclusions:
- Thermolysin treatment resulted in a statistically significant increase in keratinocyte colonies compared to trypsin.
- This method offers a more efficient approach to keratinocyte isolation for burn treatment.
