Instituto de Bioingenieria, Universidad Miguel Hernández de Elche, Elche, Spain.
Int J Toxicol. 2010 May-Jun;29(3):297-304. doi: 10.1177/1091581810366312.
Embryonic stem (ES) cells are considered an important alternative to develop in vitro screening methods for embryotoxicity. Mouse ES cells can be cultured as cell suspension aggregates termed "embryoid bodies" (EBs) in which cells start to differentiate. We have studied the expression of several genes in the presence of a wide range of concentrations of 5-fluorouracil (5-FU). This well-established embryotoxic compound completely inhibited cell viability at 200 nmol/L in monolayer cultures. At lower concentrations, 5-FU led to decrease in the expression of the alpha-fetoprotein gene, a marker of the visceral endoderm, in the EBs. However, the expression of several mesodermal gene markers was not significantly affected at these concentrations. These results suggest a high sensitivity of the visceral endoderm differentiation to 5-FU. Therefore, the quantification of the alpha-fetoprotein gene after exposure to potential embryotoxicants should be considered an additional end point in future embryotoxicity assays in vitro with ES cells.
胚胎干细胞（ES 细胞）被认为是开发体外胚胎毒性筛选方法的重要替代方法。可以将小鼠 ES 细胞培养为细胞悬浮聚集物，称为“胚状体”（EB），其中细胞开始分化。我们研究了在广泛浓度范围内存在的 5-氟尿嘧啶（5-FU）时几种基因的表达。这种成熟的胚胎毒性化合物在单层培养物中浓度为 200nmol/L 时完全抑制细胞活力。在较低浓度下，5-FU 导致胚状体中内胚层标志物甲胎蛋白基因的表达减少。然而，这些浓度下几个中胚层基因标志物的表达没有受到显著影响。这些结果表明 5-FU 对内胚层分化具有很高的敏感性。因此，在未来使用 ES 细胞进行体外胚胎毒性测定时，应考虑在接触潜在胚胎毒性物质后定量检测甲胎蛋白基因，作为另一个终点。
