Development and evaluation of an in vitro skin sensitization assay based on primary monocyte-derived dendritic cells (P6376)
| Autor: | Jochen Kuehnl, Silke Gerlach, Hendrik Reuter, Stefan Onken, Horst Wenck, Andreas Schepky |
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| Rok vydání: | 2013 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 190:201.5-201.5 |
| ISSN: | 1550-6606 0022-1767 |
| Popis: | Allergic contact dermatitis is a delayed T-cell mediated allergic response associated with relevant social and economic impacts. Until now, animal experiments (e.g. the local lymph node assay) are supplying most of the data used to assess the sensitization potential of new chemicals. However, becoming effective in March 2013, the 7th amendment to the EU Cosmetic Directive introduced an animal testing ban for the endpoint skin sensitisation for cosmetic ingredients. In vitro alternative methods are thus being actively developed. Although promising results have been obtained with cell lines, their reduced functionality and inherent genomic instability led us to reinvestigate the use of peripheral blood monocyte-derived dendritic cells (PBMDCs) for the establishment of a reliable in vitro sensitization test. We successfully solved the issues associated with the use of primary cells (optimized cytokine concentrations, incubation time, readout, pooled vs. single donors and cytoxicity) resulting in a stable and reproducible protocol. Here, we report on the adaption of the protocol to the 96-well format, allowing for a higher and cost-effective throughput of test substances. Comparative analyses of the effect of several skin sensitizers on CD86 expression display similar results for 24- and 96-well format. Upscaling of the PBMDCs advances its applicability for the reliable detection of human skin sensitizers. |
| Databáze: | OpenAIRE |
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