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Updated: Sep 3, 2025
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Generation and Culturing of Primary Human Keratinocytes from Adult Skin
Published on: December 22, 2017
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Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
Rosanna Marsella1, Kim Ahrens1, Rachel Wilkes1
1Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611, USA.
Veterinary Sciences
|July 25, 2022
Summary
Atopic dermatitis keratinocytes show lower epithelial permeability, measured by TransEpithelial Electrical Resistance (TEER). This difference is linked to their unique growth patterns, not tight junction protein variations.
Area of Science:
- Dermatology
- Cell Biology
- Biophysics
Background:
- Skin barrier dysfunction is a key feature of atopic dermatitis.
- Keratinocyte behavior in culture may reveal intrinsic differences related to the disease.
- TransEpithelial Electrical Resistance (TEER) is a standard measure of epithelial barrier function and permeability.
Purpose of the Study:
- To investigate intrinsic differences in normal and atopic dermatitis keratinocytes.
- To compare epithelial permeability and growth characteristics between normal and atopic keratinocytes.
- To determine if differences in tight junction proteins contribute to altered barrier function in atopic dermatitis.
Main Methods:
- Culturing of normal and atopic dermatitis human keratinocytes.
- Measurement of TransEpithelial Electrical Resistance (TEER) to assess barrier permeability.
- Analysis of gene expression for filaggrin and tight junction proteins (claudin, occludin, Zonula Occludens-1).
- Immunofluorescent staining and ANOVA for cell morphology and protein localization.
Main Results:
- Atopic dermatitis keratinocytes exhibited significantly lower TEER compared to normal keratinocytes.
- Atopic keratinocytes displayed distinct growth patterns, forming dome-like structures before horizontal expansion.
- Filaggrin gene expression was higher in atopic keratinocytes at confluence, indicating earlier differentiation.
- No significant differences in tight junction protein levels or continuity were observed between groups.
- Both normal and atopic keratinocytes showed increased cell size over time, a subject-dependent but not disease-dependent phenomenon.
Conclusions:
- Decreased TEER in atopic dermatitis keratinocytes is likely due to intrinsic differences in growth behavior, not alterations in tight junction proteins.
- The findings suggest that altered keratinocyte proliferation and differentiation contribute to skin barrier impairment in atopic dermatitis.
- Further research into keratinocyte growth dynamics could reveal novel therapeutic targets for atopic dermatitis.
