Construction of a Three-Dimensional Human Skin Model Consisting of Keratinocytes, Dendritic Cells and Fibroblasts and Application of This Model for Alternative Animal Testing of Immune-Sensitizing Compounds

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In order to establish in vitro evaluation of the sensitization of human skin, we attempted to make a three-dimensional human skin model consisting of three different cells, dendritic cells, keratinocytes and fibroblasts. The viability of the cells in the human skin model was observed after staining with hematoxylin and eosin. After 11-14-day incubation (horny layer was initially observed), the three-dimensional human skin model was used for experiments. Due to 2, 4-dinitrochlorobenzene (DNCB) under a non-cytotoxic dose, the keratinocytes and dendritic cells in the human skin model significantly induced IL-4 release into the incubating medium and dendritic cells induced CD86 expression. On the other hand, with sodium dodecyl sulfate (SDS; non-sensitizer), the keratinocytes and dendritic cells did not significantly induce IL-4 release and the dendritic cells did not induce CD86 expression. The results suggested that this three-dimensional human skin model with dendritic cells could be applied as an alternative to animal testing of immune-sensitizing compounds.
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