The aim of the project was to establish a fluorescence-based in vitro assay test system to map rapid non-genomic signaling cascades affected by environmentally relevant endocrine active substances.
The aim of the research project conducted by Steinbeis-Innovationszentrum Zellkulturtechnik (STZ) in Mannheim and TZW The German Water Center was to establish a fluorescence-based in vitro test system. Within this project, we successfully developed sensor cell lines to efficiently monitor rapid non-genomic signaling cascades influenced by environmentally relevant endocrine-active substances. These sensor cell lines were created by genetically modifying G-Protein coupled estrogen receptor (GPER1) expressing cells, essentially turning them into artificial fluorescent signalosomes. Sensor cells were equipped with reporter units allowing the monitoring of cellular cAMP or Ca2+ intracellular concentration changes. Using this sensor cell lines we developed fluorescence-based in vitro assays, enabling us to measure cAMP or Ca2+ modulation with high time resolution. We characterized the GPER1 sensor cells using both agonists and antagonist compounds. Our newly developed sensor cell-based in vitro assays were then employed to investigate potentially endocrine-disrupting compounds. Additionally, we used these assays to evaluate the influents and effluents from advanced treatment processes of wastewater treatment plants.
Publication
Bernhard, K.; Stahl, C.; Martens, R.; Köhler, H-R.; Triebskorn, R.; Scheurer, M.; Frey, M.: Two novel real time cell-based assays quantify beta-blocker and NSAID specific effects in effluents of municipal wastewater treatment plants. Water Research 115, 74-83 (2017)
