Protocol for the Isolation of Stratum Corneum from Pig Ear Skin: Evaluation of the Trypsin Digestion Conditions
Abstract
:1. Introduction
2. Materials and Equipment
3. Experimental Design
3.1. Study Aims
- i.
- Design and evaluation of different experimental parameters, varying the trypsin concentration, the temperature, the time of digestion and the freezing of the skin prior to digestion;
- ii.
- Study of the influence of each set of conditions on the isolated SC regarding the visual aspect, histological features and permeability properties;
- iii.
- Characterization of the SC model isolated in the selected optimal conditions by assessment of the histological morphology and storage stability.
3.2. Preliminary Steps: Preparation of Pig Ear Skin
3.3. Design of the Methodologies and Visual Inspection of the Skin after Trypsin Digestion
3.4. Evaluation of the Calcein Permeability
3.5. Histological Characterization
4. Results
4.1. Characterization of the Model Obtained in the Selected Conditions: Histological Analysis
4.2. Characterization of the Model Obtained in the Selected Conditions Storage Stability
5. Limitations
6. Concluding Remarks
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Condition | Incubation Time | Temperature | Trypsin Concentration | Pre- Freezing |
---|---|---|---|---|
A | 4 h + O/N 1 (total ~ 20 h) | 4 °C | 0.1% | No |
B | 4 h + O/N 1 (total ~ 20 h) | 4 °C | 0.1% | Yes, 48 h |
C | 3 h | 37 °C | 0.1% | No |
D | 3 h | 37 °C | 0.05% | No |
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Moniz, T.; Lima, S.A.C.; Reis, S. Protocol for the Isolation of Stratum Corneum from Pig Ear Skin: Evaluation of the Trypsin Digestion Conditions. Methods Protoc. 2021, 4, 80. https://doi.org/10.3390/mps4040080
Moniz T, Lima SAC, Reis S. Protocol for the Isolation of Stratum Corneum from Pig Ear Skin: Evaluation of the Trypsin Digestion Conditions. Methods and Protocols. 2021; 4(4):80. https://doi.org/10.3390/mps4040080
Chicago/Turabian StyleMoniz, Tânia, Sofia A. Costa Lima, and Salette Reis. 2021. "Protocol for the Isolation of Stratum Corneum from Pig Ear Skin: Evaluation of the Trypsin Digestion Conditions" Methods and Protocols 4, no. 4: 80. https://doi.org/10.3390/mps4040080