Further development of test methods for endocrine active substances

Ca2+ Signalosom (Image: Manfred Frey /STZ Mannheim)

The aim of the project is 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 is to establish a fluorescence-based in vitro test system. This system is to be used to map rapid non-genomic signal cascades that are influenced by environmentally relevant endocrine active substances. For this purpose, STZ is developing a sensor cell line that is genetically modified to induce an artificial signalosome. After induction, the cell line to be developed expresses various proteins which, together with cellular components, form a functional signaling unit localized at the cell membrane, a signalosome. The core of this signalosome is formed by recombinant membrane-bound estrogen receptors. In addition, the cell is equipped with a reporter unit that matches the signalosome and, after binding an endocrine-active compound, converts the specific signal generated at the signalosome into a quantifiable fluorescence signal. Based on this sensor cell, a new in vitro bioassay will then be developed to quantitatively detect estrogen-active compounds via their effects. Specificity and sensitivity of the new in-vitro bioassay will be tested with selected control substances. In the context of a practical suitability test, sample processing procedures of polluted surface water samples will be established. The samples will then be characterized in parallel in the in-vitro bioassay and by chemical analysis mass spectrometric detection by TZW. The data obtained here should demonstrate the practical suitability of the new in-vitro bioassay for use in the context of an assessment of surface water qualities with regard to their estrogenic effect potentials for the aquatic environment.