shortening and improving the embryonic stem cell test through the use of gene biomarkers of differentiation缩短和改善测试通过使用胚胎干细胞基因生物标记的区别.pdfVIP

Hindawi Publishing Corporation Journal of Toxicology Volume 2011, Article ID 286034, 8 pages doi:10.1155/2011/286034 Research Article Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation Andrea C. Romero, Eugenio Vilanova, and Miguel A. Sogorb Unidad de Toxicologıa y Seguridad Quımica, Instituto de Bioingenierıa, Universidad Miguel Hernandez de Elche, ´ ´ ´ ´ Avenida de la Universidad s/n, 03202 Elche, Spain Correspondence should be addressed to Miguel A. Sogorb, msogorb@umh.es Received 11 May 2011; Revised 30 June 2011; Accepted 1 July 2011 Academic Editor: Yujian James Kang Copyright © 2011 Andrea C. Romero et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The embryonic Stem cell Test (EST) is a validated assay for testing embryotoxicity in vitro. The total duration of this protocol is 10 days, and its main end-point is based on histological determinations. It is suggested that improvements on EST must be focused toward molecular end-points and, if possible, to reduce the total assay duration. Five days of exposure of D3 cells in monolayers under spontaneous differentiation to 50 ng/mL of the strong embryotoxic 5-fluorouracil or to 75 μg/mL of the weak embryotoxic 5,5-diphenylhydeantoin caused between 20 and 74% of reductions in the expression of the following genes: Pnpla6, Afp, Hdac7, Vegfa, and Nes. The exposure to 1 mg/mL of nonembryotoxic saccharin only caused statistically significant reductions in the expression of Nes. These exposure