Optimization and prevalidation of the in vitro ERalpha CALUX method to test estrogenic and antiestrogenic activity of compounds.
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.
van der Burg
Bart
B
BioDetection Systems BV, Science Park 406, Amsterdam, The Netherlands. bart.van.der.burg@bds.nl
Winter
Roos
R
Weimer
Marc
M
Berckmans
Pascale
P
Suzuki
Go
G
Gijsbers
Linda
L
Jonas
Arjen
A
van der Linden
Sander
S
Witters
Hilda
H
Aarts
Jac
J
Legler
Juliette
J
Kopp-Schneider
Annette
A
Bremer
Susanne
S
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
D052244:Endocrine Disruptors; D004965:Estrogen Antagonists; D047628:Estrogen Receptor alpha; D004967:Estrogens; D008156:Luciferases