Critical parameters in the MCF-7 cell proliferation bioassay (E-Screen).
The MCF-7 cell proliferation bioassay has grown in popularity as a rapid test for detecting potentially oestrogenic compounds. Several MCF-7 cell sublines with different sensitivities to oestrogens are currently used, with maximal proliferation responses ranging from two- to 10-fold above those of hormone-free controls. In the highly responsive MCF-7 BUS cell line, we evaluated critical assay parameters for test performance, including growth conditions, initial seeding densities and differences in growth stimulation in medium containing human serum or fetal calf serum as well as appropriate solvents for oestrogen-mimicking compounds. Modifications significantly reduced the labour-intensive steps and overall assay costs without affecting the sensitivity of the assay. Using this optimized test regimen, the responsiveness of treated MCF-7 BUS cells was consistently increased up to 11-fold over hormone-free controls. The specificity was characterized by examining the effects of oestradiol-17beta, the anti-oestrogen ICI 182,780, and dieldrin, a recognized xeno-oestrogen. The improved proliferation bioassay will be a useful tool in identifying potential xeno-oestrogens.
Rasmussen
Thomas Høj
TH
Department of Environmental Medicine, Institute of Public Health, University of Southern Denmark, DK-5000 Odense C, Denmark. thoj@health.sdu.dk
Nielsen
Jesper Bo
JB
D001681:Biological Assay / Q000379:methods; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects*; D003470:Culture Media; D004026:Dieldrin / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000494:pharmacology*; D005260:Female; D000077267:Fulvestrant; D006801:Humans; D009376:Neoplasms, Hormone-Dependent / Q000378:metabolism / Q000473:pathology; D011960:Receptors, Estrogen / Q000187:drug effects / Q000378:metabolism; D012235:Rhodamines; D012997:Solvents; D014407:Tumor Cells, Cultured
D003470:Culture Media; D004968:Estrogens, Non-Steroidal; D011960:Receptors, Estrogen; D012235:Rhodamines; D012997:Solvents; D000077267:Fulvestrant; C022027:lissamine rhodamine B; D004958:Estradiol; D004026:Dieldrin