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Embryonic stem cells (ES cells), pluripotent cells derived from the inner cell mass of blastocysts, differentiate in vitro into a variety of cell types representing all three germ layers. They therefore constitute one of the most promising in vitro tools for developmental toxicology. To assess the developmental toxicity of chemicals using ES cells easily, identification of effective marker genes is a high priority. We report here altered gene expression during ES cell differentiation into myocardiac and neural cells on treatment with some embryotoxic and non-embryotoxic chemicals. Decreases in several undifferentiated markers such as Oct3/4 and Nanog, and elevated expression of genes associated with heart development or the central nervous system, respectively, were found on microarray analysis. Under differentiation of ES cells into myocardic cells, 107 genes were substantially up-regulated. Decrease in the expression of 13 genes of these (Hand1, Pim2, Tbx20, Myl4, Myl7, Hbb-bh1, Hba-a1, Col1a2, Hba-x, Cmya1, Pitx2, Smyd1 and Adam19) was observed specifically by embryotoxic chemicals. Of the 107 genes up-regulated under differentiation into neurons, 22 genes (Map2, Cpe, Marcks, Ptbp2, Sox11, Tubb2b, Vim, Arx, Emx2, Pax6, Basp1, Ddr1, Ndn, Sfrp, Ttc3, Ubqln2, Six3, Dcx, L1cam, Reln, Wnt1 and Nnat) showed reduced expression specifically by embryotoxic chemicals. Almost all gene sets identified in this study are known to be indispensable for differentiation and development of heart and brain tissues, and thus may serve in early detection or prediction of embryotoxicity of chemicals in vitro.

journal of toxicological sciences

Analysis of altered gene expression specific to embryotoxic chemical treatment during embryonic stem cell differentiation into myocardiac and neural cells