Effects of environmental chemicals on development of neuronal cell lineages derived from mouse ES cells

This study provides a comprehensive evaluation of changes in gene expression during treatment with environmental chemicals in embryonic bodies derived from mouse embryonic stem (ES) cells. ES cells were maintained under the feeder cells in phenol red-free DMEM Medium (supplemented with 15% FBS, 100uM Non-essential amino acids (NEAA), 1000U/ml Leukemia inhibitory factor(LIF), 100uM 2-mercaptoethanol(2-ME), and 0.5% penicillin/streptomycin). Cells were gently washed in warm PBS and transferred to phenol red-free DMEM Medium (supplemented with 15% KnockOut Serum Replacement, 100uM NEAA, 100uM 2-ME, and 0.5% penicillin/streptomycin) in Microsphere array (MSA300F) and exposed with low and high levels of each chemical just after removing LIF from the culture media. Prior to collection, cells were washed in warm PBS, resuspended and briefly incubated in the QIAGEN RLT buffer, and finally collected in triplicate at 48 hours. Following RNA isolation, the best RNA yields for each replicate set was selected for target preparation and microarray processing. 25 total samples: 1 vehcle, 12 chemicals and two doses of each chemical [Vehicle Control (0.1% dimethyl sulfoxide), estradiol (1nM and 10nM), dehydrotestosterone (1nM and 10nM) bisphenol A (1pM and 100pM), triiodothyronine (1nM and 10nM), dexamethasone (1nM and 10nM), 2,3,7,8-tetrachlorodibenzo-p-dioxin (1nM and 10nM), 4(OH)-2’3,3’4’-5’-pentachlo biphenyls (1nM and 10nM), thalidomide (100nM and 0.01mM), bis(2-ethylhexyl)phthalate (0.001mM and 0.1mM), permethrin (100nM and 0.01mM), cyclopamine (100nM and 0.01mM), methoprene acid (0.001mM and 0.1mM)]; 1 Timepoint [48 hours].