title: A sensitive sensor cell line for the detection of oxidative stress responses in cultured human keratinocytes.
abstract: In the progress of allergic and irritant contact dermatitis, chemicals that cause the generation of reactive oxygen species trigger a heat shock response in keratinocytes. In this study, an optical sensor cell line based on cultured human keratinocytes (HaCaT cells) expressing green fluorescent protein (GFP) under the control of the stress-inducible HSP70B' promoter were constructed. Exposure of HaCaT sensor cells to 25 µM cadmium, a model substance for oxidative stress induction, provoked a 1.7-fold increase in total glutathione and a ~300-fold induction of transcript level of the gene coding for heat shock protein HSP70B'. An extract of Arnica montana flowers resulted in a strong induction of the HSP70B' gene and a pronounced decrease of total glutathione in keratinocytes. The HSP70B' promoter-based sensor cells conveniently detected cadmium-induced stress using GFP fluorescence as read-out with a limit of detection of 6 µM cadmium. In addition the sensor cells responded to exposure of cells to A. montana extract with induction of GFP fluorescence. Thus, the HaCaT sensor cells provide a means for the automated detection of the compromised redox status of keratinocytes as an early indicator of the development of human skin disorders and could be applied for the prediction of skin irritation in more complex in vitro 3D human skin models and in the development of micro-total analysis systems (µTAS) that may be utilized in dermatology, toxicology, pharmacology and drug screenings.
authors: Department of Medical Engineering and Biotechnology, University of Applied Sciences Jena, Carl-Zeiss-Promenade 2, 07745 Jena, Germany. ute.hofmann@fh-jena.de.
authors: Ute
authors: U
authors: Hofmann
authors: Department of Medical Engineering and Biotechnology, University of Applied Sciences Jena, Carl-Zeiss-Promenade 2, 07745 Jena, Germany. melanie.priem@fh-jena.de.
authors: Melanie
authors: M
authors: Priem
authors: Department of Medical Engineering and Biotechnology, University of Applied Sciences Jena, Carl-Zeiss-Promenade 2, 07745 Jena, Germany. christine.bartzsch@fh-jena.de.
authors: Christine
authors: C
authors: Bartzsch
authors: Department of Pharmaceutical Biology, Institute of Pharmacy, Friedrich-Schiller-University of Jena, Semmelweisstraße 10, 07743 Jena, Germany. t.winckler@uni-jena.de.
authors: Thomas
authors: T
authors: Winckler
authors: Department of Medical Engineering and Biotechnology, University of Applied Sciences Jena, Carl-Zeiss-Promenade 2, 07745 Jena, Germany. feller@fh-jena.de.
authors: Karl-Heinz
authors: KH
authors: Feller
mesh_terms: D001681:Biological Assay / Q000295:instrumentation*; D060846:Cell Engineering / Q000379:methods; D002460:Cell Line; D004867:Equipment Design; D019544:Equipment Failure Analysis; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects* / Q000473:pathology / Q000502:physiology*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000032:analysis* / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*
chemical_list: D018840:HSP70 Heat-Shock Proteins; C491129:HSPA7 protein, human; D017382:Reactive Oxygen Species