A Panel of Quantitative Calux® Reporter Gene Assays for Reliable High‐Throughput Toxicity Screening of Chemicals and Complex Mixtures

Innovative testing in reproductive toxicology—The ChemScreen experience

“…In 2014, the annual prize for best manuscript in Reproductive Toxicology, awarded by Elsevier and juried by the European Teratology Society, was given for a publication presenting the ChemScreen battery approach, illustrating the appreciation for this line of research in the scientific community [10]. The present special issue of Reproductive Toxicology brings together a series of key original contributions of the ChemScreen output, adding to a growing list of scientific publications from the project [10][11][12][13][14].…”

A high throughput screening system for predicting chemically-induced reproductive organ deformities

“…In addition a range of highly selective human U2-OS cell based lines were used including doubly (i.e. reporter gene and receptor) transfected ERalpha-, PR (progesterone receptor)-, GR (glucocorticoid receptor)-, AR (androgen receptor) CALUX lines [11], and an extension of the panel with peroxisome proliferator activated receptor PPAR␣-and PPAR␥ CALUX assays [58,59], TR (thyroid receptor)␤-, RAR (retinoic acid receptor)-, ER (estrogen receptor)␤ and LXR (liver X receptor) CALUX [11]. This screening panel was completed with a range of U2-OS-based singly transfected lines expressing a reporter gene only, which are designed to selectively measure the activity of main intracellular signaling pathways.…”

“…The CALUX battery of stable reporter gene assays [11] comprised assays and test conditions that were selected from a larger panel of CALUX cells because of their possible relevance to reproductive toxicity testing. A starting point of the current panel was a range of reporter gene assays for nuclear hormone receptors.…”

“…A second part of our strategy was to use assays that are highly selective for specific molecular pathways, as has been typically the approach in the setup of the CALUX battery of U2-OS based reporter gene assays [10,11]. This is attained by using minimal promoter elements coupled to multimerized, highly selective response promoter elements in these assays driving luciferase expression.…”

Mining into interspecific bacterial interactions

“…The mammalian CALUX® reporter assays, DR-CALUX (dioxin-like activity), Nrf2-CALUX, ERα-CALUX (estrogenic activity), AR-CALUX (androgenic activity) and p53-CALUX (genotoxicity) were conducted as describe in (van der Burg et al, 2013;van der Burg et al, 2015). The 2,5-bis(1-methylethyl)-pyrazine formulation was dissolved at 0.8 % (DR), 0.1 % (ERα, AR and cytotoxicity) and 1 % (p53 and Nrf2) in 6 well plates in DMEM/F-12 medium (Life Technologies), while shaking at room temperature for 10 minutes and 200 rpm at room temperature and diluted accordingly.…”