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  4. A coculture system composed of THP-1 cells and 3D reconstructed human epidermis to assess activation of dendritic cells by sensitizing chemicals after topical exposure
 
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2019
Journal Article
Title

A coculture system composed of THP-1 cells and 3D reconstructed human epidermis to assess activation of dendritic cells by sensitizing chemicals after topical exposure

Abstract
A key event of the adverse outcome pathway for skin sensitization is the activation of dendritic cells (DC). To be most close to the in vivo situation, we combined for the first time reconstructed human epidermis (RHE) in coculture with THP-1 cells, as surrogate for DC. THP-1 cells were placed underneath RHE (SkinEthicTM, OS-REp). Cell activation was measured by analyzing cell surface expression of co-stimulatory molecules CD86 and CD40, adhesion molecule CD54 and of human leukocyte antigen class II-related subtypes (HLA-DR) on THP-1 cells by flow cytometry. Both models were suitable to measure DC activation but basal CD54 levels are significantly increased compared to THP-1 cells in monoculture for OS-REp. Chemical-induced activation of THP-1 cells was investigated after topical exposure on SkinEthicTM. As proof of concept we analyzed three sensitizers and lactic acid (non-sensitizer). We observed differential, dose dependent levels of CD86 and/or CD54 on THP-1 cells in response to the skin sensitizers. We conclude that the RHE/THP-1 coculture using topical exposure complements our HaCaT/THP-1 coculture (COCAT) based on a submersed exposure and may allow the analysis of DC activation by various kind of test items including chemicals with pronounced lipophilicity, mixtures and possibly finished products.
Author(s)
Schellenberger, Mario T.
Bock, Udo
Hennen, Jennifer
Groeber-Becker, Florian Kai  
Fraunhofer-Institut für Silicatforschung ISC  
Walles, Heike
Fraunhofer-Institut für Silicatforschung ISC  
Blömeke, Brunhilde
Journal
Toxicology in vitro  
DOI
10.1016/j.tiv.2019.02.002
Language
English
Fraunhofer-Institut für Silicatforschung ISC  
Keyword(s)
  • epidermis

  • Cokultur

  • Zellkultur

  • Haut

  • dendritische Zelle

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