Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation
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 po...
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| Format: | Article |
| Language: | English |
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Wiley
2011-01-01
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| Series: | Journal of Toxicology |
| Online Access: | http://dx.doi.org/10.1155/2011/286034 |
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| author | Andrea C. Romero Eugenio Vilanova Miguel A. Sogorb |
| author_facet | Andrea C. Romero Eugenio Vilanova Miguel A. Sogorb |
| author_sort | Andrea C. Romero |
| collection | DOAJ |
| description | 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 exposures reduced cell viability of D3 cells by 15, 28, and 34%. We applied these records to the mathematical discriminating function of the EST method to find that this approach is able to correctly predict the embryotoxicity of all three above-mentioned chemicals. Therefore, this work proposes the possibility of improve EST by reducing its total duration and by introducing gene expression as biomarker of differentiation, which might be very interesting for in vitro risk assessment embryotoxicity. |
| format | Article |
| id | doaj-art-d2f021065d1246938b3da2137fede014 |
| institution | OA Journals |
| issn | 1687-8191 1687-8205 |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Toxicology |
| spelling | doaj-art-d2f021065d1246938b3da2137fede0142025-08-20T02:19:50ZengWileyJournal of Toxicology1687-81911687-82052011-01-01201110.1155/2011/286034286034Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of DifferentiationAndrea C. Romero0Eugenio Vilanova1Miguel A. Sogorb2Unidad de Toxicología y Seguridad Química, Instituto de Bioingeniería, Universidad Miguel Hernández de Elche, Avenida de la Universidad s/n, 03202 Elche, SpainUnidad de Toxicología y Seguridad Química, Instituto de Bioingeniería, Universidad Miguel Hernández de Elche, Avenida de la Universidad s/n, 03202 Elche, SpainUnidad de Toxicología y Seguridad Química, Instituto de Bioingeniería, Universidad Miguel Hernández de Elche, Avenida de la Universidad s/n, 03202 Elche, SpainThe 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 exposures reduced cell viability of D3 cells by 15, 28, and 34%. We applied these records to the mathematical discriminating function of the EST method to find that this approach is able to correctly predict the embryotoxicity of all three above-mentioned chemicals. Therefore, this work proposes the possibility of improve EST by reducing its total duration and by introducing gene expression as biomarker of differentiation, which might be very interesting for in vitro risk assessment embryotoxicity.http://dx.doi.org/10.1155/2011/286034 |
| spellingShingle | Andrea C. Romero Eugenio Vilanova Miguel A. Sogorb Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation Journal of Toxicology |
| title | Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation |
| title_full | Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation |
| title_fullStr | Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation |
| title_full_unstemmed | Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation |
| title_short | Shortening and Improving the Embryonic Stem Cell Test through the Use of Gene Biomarkers of Differentiation |
| title_sort | shortening and improving the embryonic stem cell test through the use of gene biomarkers of differentiation |
| url | http://dx.doi.org/10.1155/2011/286034 |
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