title: Optimization and prevalidation of the in vitro ERalpha CALUX method to test estrogenic and antiestrogenic activity of compounds.
abstract: Estrogenicity of chemicals has received significant attention and is linked to endocrine-disrupting activities. However, there is a paucity of validated methods to assess estrogenicity in vitro. We have established a robust method to test estrogenic and antiestrogenic activity of compounds in vitro, as an alternative to using animal models such as the uterotrophic assay. To this end we optimized protocols to be used in combination with CALUX reporter gene assays and carried out an in house prevalidation, followed by two rounds of tests to establish transferability. Problems in the initial test with transferability were solved by isolation of a novel cell clone of the ERalpha CALUX line with greatly improved stability and luciferase levels. This cell line proved to be a very suitable and reliable predictor of estrogenicity of chemicals and was able to readily rank a range of chemicals on the basis of their EC50 values.
authors.affiliation: BioDetection Systems BV, Science Park 406, Amsterdam, The Netherlands. bart.van.der.burg@bds.nl
authors.forename: Bart
authors.initials: B
authors.lastname: van der Burg
authors.forename: Roos
authors.initials: R
authors.lastname: Winter
authors.forename: Marc
authors.initials: M
authors.lastname: Weimer
authors.forename: Pascale
authors.initials: P
authors.lastname: Berckmans
authors.forename: Go
authors.initials: G
authors.lastname: Suzuki
authors.forename: Linda
authors.initials: L
authors.lastname: Gijsbers
authors.forename: Arjen
authors.initials: A
authors.lastname: Jonas
authors.forename: Sander
authors.initials: S
authors.lastname: van der Linden
authors.forename: Hilda
authors.initials: H
authors.lastname: Witters
authors.forename: Jac
authors.initials: J
authors.lastname: Aarts
authors.forename: Juliette
authors.initials: J
authors.lastname: Legler
authors.forename: Annette
authors.initials: A
authors.lastname: Kopp-Schneider
authors.forename: Susanne
authors.initials: S
authors.lastname: Bremer
mesh_terms: D000826:Animal Testing Alternatives; D001681:Biological Assay / Q000379:methods* / Q000592:standards; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D052244:Endocrine Disruptors / Q000494:pharmacology*; D004965:Estrogen Antagonists / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics; D004967:Estrogens / Q000494:pharmacology*; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics; D015203:Reproducibility of Results
chemical_list: D052244:Endocrine Disruptors; D004965:Estrogen Antagonists; D047628:Estrogen Receptor alpha; D004967:Estrogens; D008156:Luciferases