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The LUMI-CELL® estrogen receptor (ER) assay measures whether, and to what extent, a substance induces or inhibits transcriptional activation
(TA) activity via ER-mediated pathways in recombinant BG-1Luc4E2 cells. The BG-1Luc4E2 cell line was derived
from BG-1 immortalized adenocarcinoma cells that endogenously express ER and that have been stably
transfected with the plasmid pGudLuc7.ERE. This plasmid contains four copies of a synthetic oligonucleotide
containing the estrogen response element upstream of the mouse mammary tumor viral promoter and the firefly
luciferase gene. BG-1Luc4E2 cells express luciferase activity in response to estrogen and estrogen-like substances.
ICCVAM recently completed a protocol standardization study for the LUMI-CELL® ER assay which:
- Standardized procedures for using the assay to identify ER agonists and antagonists
- Standardized procedures for a quantitative assessment of cell viability for use with the agonist
and antagonist assays
- Developed two GLP-compliant protocols: one for the agonist assay, and one for the antagonist assay
- Established a historical database for reference standards and controls used in the agonist and antagonist
assay protocols
- Demonstrated the adequacy of the standardized protocols for detecting ER agonists or antagonists using
eight substances covering a range of ER agonist and antagonist activities, respectively.
ICCVAM is currently conducting an international validation study of the LUMI-CELL® ER assay using the standardized
protocols in conjunction with the European Centre for the Validation of Alternative Methods (ECVAM) and the Japanese
Center for the Validation of Alternative Methods (JaCVAM). The specific objectives of this validation study are to:
- Further standardize and optimize the LUMI-CELL® ER assay using the agonist and antagonist protocols to
test the 78 ICCVAM recommended substances for the validation of in vitro ER TA test methods in three
laboratories, one in Europe, Japan, and the U.S, to maximize test method reliability (intralaboratory
repeatability, intra- and inter-laboratory reproducibility)
- Use the results from the testing of the 78 ICCVAM recommended substances to develop a high quality in vitro
ER TA database that can be used to characterize the extent to which other individual in vitro endocrine
disruptor test methods (or test method batteries) might be used to further reduce the expected requirements for
animal use in the screening of potential endocrine disruptors
Test Method Nomination: Submission of XDS’s LUMI-CELL® ER High-Throughput System for Screening
Estrogen-Like Chemicals for Review by ICCVAM (February 2005)
- Submission Package [PDF]
- Letter Accompanying Submission Package from Drs. George Clark and John Gordon, XDS, Inc. [PDF]
Draft Evaluation: Nomination of the LUMI-CELL® ER High-Throughput System for Screening Estrogen-Like Chemicals
for Validation Studies (August 2004) [PDF]
Protocols for the International Validation Study:
- LUMI-CELL® ER Agonist Protocol (updated October 2008) [PDF]
- LUMI-CELL® ER Antagonist Protocol (updated October 2008) [PDF]
Links to Home Pages of Laboratories Participating in the Conduct of the LUMI-CELL® ER Assay Validation Study
The links listed below connect to pages outside the NICEATM-ICCVAM website. These links are for the convenience of visitors
to this page, who may find them useful. NICEATM/ICCVAM is not responsible for the availability or content of these external sites,
nor do we endorse, warrant or guarantee any information or any products or services described or offered at these other sites.
Xenobiotic Detection Systems, Inc.
Hiyoshi Corporation
ECVAM Laboratories
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