EPA ToxCast Assays 01aug2001
Contractor or CollaboratorAssay Type# Assays orEndpointsReferences
ACEA Biosciences, Inc.real-time cell electronic sensing(RT-CES)1Xing et al 2006. Microelectronic cell sensorassay for detection of cytotoxicity andprediction of acute toxicity. Toxicol In Vitro20:995-1004; www.aceabio.comAttagene, Inc.transcription factor activities;reporter gene expression67US Patent Application 20060160108Populations of reporter sequences andmethods of their use; www.attagene.comBioSeek Inc.complex primary human cell-based87Berg et al 2006. Characterization of compound mechanisms and secondaryactivities by BioMAP analysis. J PharmacolToxicol Methods 53:67-74;www.bioseekinc.comCellumen, Inc.cellular high content screening(HCS)11Giuliano et atl 2006. Systems cell biologybased on high-content screening. MethodsEnzymol 414:601-19; www.cellumen.comCompound Focus, Inc. / BioFocus DPIchemical procurement andhandlingn/awww.sbsonline.org/publications/news/issues/ 2005_12/index.php ; www.biofocus.comExpression Analysis, Inc.in vitro genomics; geneexpression>20,000Shi et al 2006. The MicroArray QualityControl (MAQC) project shows inter- andintraplatform reproducibility of geneexpression measurements. Nat Biotechnol.2006 Sep;24(9):1151-61;www.expressionanalysis.comIn Vitro ADMETLaboratories (IVAL), LLC.cell culture and co-culture1Li AP 2007. Human hepatocytes: isolation,cryopreservation and applications in drugdevelopment. Chem Biol Interact 168:16-29;www.invitroadmet.comNovaScreen BiosciencesCorp.biochemical and cellular highthroughput screeening (HTS)240www.novascreen.comPhylonix Pharmaceuticals,Inc.zebrafish developmental toxicity13Parng et al 2007. Neurotoxicity assessmentusing zebrafish. J Pharmacol ToxicolMethods 55:103-112; www.phylonix.comNIH Chemical GenomicsCenterbiochemical and cellular highthroughput screeening (HTS)>10Inglese et al 2006. Quantitative high-throughput screening: a titration-basedapproach that efficiently identifies biologicalactivities in large chemical libraries. Proc NatlAcad Sci USA 103:11473-8;www.ncgc.nih.gov
ToxCast Phase 1 Assays01aug2007
Contractor orCollaboratorAssaySpecies/CellInitialConcentrationsScreened
ACEAReal-time cell electronic sensing (RT-CES)providing a kinetic "Cell Index" based onquantitative impedance measurementsindicative of cell number, viability,morphology, and cytoskeletal dynamics.Human lung carcinoma cell lineA5498 pointconcentrationresponseATTAGENECisCis Reporter Transcription Unit (RTU)assays for 43 distinct transcription factorsHuman liver carcinoma cell lineHepG2, stably transfected 25 uMTransTrans RTU assays for 24 distinct humannuclear receptorsHuman liver carcinoma cell lineHepG2, stably transfected 25 uM
gal4-ERRa Estrogen-related receptor alpha (NR3B1)gal4-PXR Pregnane X receptor (NR1I2)gal4-PPARg Peroxisome proliferator activated receptor gamma (NR1C3)gal4-RXRb Retinoid X receptor (NR2B2)gal4-VDR Vitamin D3 receptor (NR1I1)gal4-GR Glucocorticoid receptor (NR3C1)gal4-ERa Estrogen receptor alpha (NR3A1)gal4-ERRg Estrogen-related receptor gamma (NR3B3)gal4-THRa Thyroid hormone receptor alpha (NR1A1)gal4-GCNF Germ cell nuclear factor 1 (NR6A1)gal4-PPARd1 Peroxisome proliferator activated receptor delta (NR1C2)gal4-FXR Farnesoid X-activated receptor (NR1H4)gal4-RARa Retinoic acid receptor alpha (NR1B1)gal4-LXRa Liver X receptor alpha (NR1H3)gal4-LXRb Liver X receptor beta (NR1H2)gal4-CAR Constitutive androstane receptor (NR1I3)gal4-RXRa Retinoid X receptor alpha (NR2B1)
Cell differentiation- HNF6, Tcf4/b-cat, PPR, FoxA, SMADs, Pax, ROR, GLI,GATA, CEBP, RAR, FoxO, Sox, Sp1Cell adhesion- Tcf4/b-cat, NFI, Nf-kB, SMADs, STATsCell proliferation- SMADS, E2F, c-Myc, p53, Egr-1, Ap1, STATs, Nf-kB,CREBStress response- p53, Nf-kB, MTF, HSF, Xbp1, HIF1a, Ets, CREB, Egr-1,STATs, PXRImmune response- Nf-kB, IRFs, STATs, Ap1Glucose and lipid homeostasis- PPR, CREB, LXR, SREBP, ROR, CEBP, FXROxidative responses- NF-E2, NRF1, HIF1aAdipogenesis- PPR, GR, CREB, CEBPHematopoiesis- HIF1a, MybMorphogenesis- NFI, Oct, Pax, Sox, ER, Ap-2Controls- viral promoters (SV40 and CMV) and minimal promoters (TATA-box,Tk-minimal)
ToxCast Phase 1 Assays01aug2007gal4-RORg RAR-related orphan receptor gamma (NR1F3)gal4-PPARa Peroxisome proliferator activated receptor alpha (NR1C1)gal4-SF1 Steroidogenic factor-1 (NR5A1)gal4-NURR1 Nur-related protein 1 (NR4A2)gal4-ATF2 Activating transcription factor 2 (ATF2)gal4-ELk1 Ets-like protein-1 (ELK1)gal4-cJun Cellular homolog of v-jun oncogene (JUN)BIOSEEK3C-9, 3 stimuli, 11 endpointsPrimary Human Endothelialcells (HUVEC), passage 4
4 pointconcentrationresponse
4H-9, 2 stimuli, 7 endpointsHUVEC, passage 4
4 pointconcentrationresponse
LPS-9, 1 stimuli, 10 endpointsHUVEC, passage 4; HumanPeripheral BloodMononuclear Cells,unpassaged
4 pointconcentrationresponse
SAg-9, 1 stimuli, 9 endpointsHUVEC, passage 4; HumanPeripheral BloodMononuclear Cells,unpassaged
4 pointconcentrationresponse
BE3C-9, 3 stimuli, 9 endpointsPrimary human bronchialepithelial cells, passage 3
4 pointconcentrationresponse
BEF4T-9, 2stimuli, 10 endpointsPrimary human bronchialepithelial cells, passage 3;Primary human dermalfibroblasts, passage 5
4 pointconcentrationresponse
HDF3CGF-9, 6 stimuli, 11 endpointsPrimary human dermalfibroblasts, passage 5
4 pointconcentrationresponse
KF3CT-9, 4 stimuli, 8 endpointsPrimary human primarykeratinocytes, passage 3;Primary human dermalfibroblasts, passage 5
4 pointconcentrationresponse
HSM3C-9, 3 stimuli, 12 endpointsHUVEC, passage 4; PrimaryHuman Vascular smoothmuscle cells, passage 5
4 pointconcentrationresponse
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Environmental Protection Agency: ToxCast Assays 01aug2007
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