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Updated: Jun 10, 2026
21:16
An In Vitro Skin Irritation Test (SIT) using the EpiDerm Reconstructed Human Epidermal (RHE) Model
Published on: July 13, 2009
Evaluation in vitro of epidermal cell keratinization
R K Ward1, P N Nation, M Maxwell
1FRAME Alternatives Laboratory, Department of Human Morphology, University of Nottingham, Nottingham NG7 2UH, UK.
Summary
Rhodamine B binding to SVK-14 cells did not reliably indicate epidermal keratinization. Further studies with normal human keratinocytes are needed to validate this in vitro method.
Area of Science:
- Dermatology
- Cell Biology
- Biochemistry
Background:
- Rhodamine B is a histopathological stain for keratinization.
- Investigating in vitro indicators for epidermal keratinization is crucial.
- SVK-14 cells offer a potential alternative to primary keratinocytes.
Purpose of the Study:
- To evaluate Rhodamine B as an in vitro indicator of epidermal keratinization.
- To assess the influence of calcium and vitamin D on keratinization in SVK-14 cells.
- To determine the suitability of SVK-14 cells for keratinization studies.
Main Methods:
- Cultured SVK-14 cells were exposed to varying calcium levels and 1,25(OH)2 vitamin D3.
- Alamar blue conversion measured cellular reductive potential.
- Rhodamine B binding relative to total protein indicated keratinization.
Main Results:
- 1,25(OH)2 vitamin D3 exposure did not increase Rhodamine B binding in SVK-14 cells.
- Calcium concentration did not affect Rhodamine B binding.
- Variability in Rhodamine B dye-binding complicated data interpretation.
Conclusions:
- Rhodamine B binding to SVK-14 cells is not a reliable indicator of epidermal keratinization.
- SVK-14 cells may have limited differentiation capacity, requiring further investigation.
- Normal human keratinocytes are recommended for validating this in vitro endpoint.
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