Reliable in vitro models to assess developmental toxicity of drugs and chemicals would lead to improvement in fetal safety and a reduced cost of drug development. The validated embryonic stem cell test (EST) uses cardiac differentiation of mouse embryonic stem cells (mESCs) to predict in vivo developmental toxicity, but does not take into account the stage-specific patterning of progenitor populations into anterior (ventricular) and posterior (atrial) compartments. In this study, we generated a novel dual reporter mESC line with fluorescent reporters under the control of anterior and posterior cardiac promoters. Reporter expression was observed in nascent compartments in transgenic mouse embryos, and mESCs were used to develop differentiation assays in which chemical modulators of Wnt (XAV939: 3, 10 µM), retinoic acid (all-trans retinoic acid: 0.1, 1, 10 µM; 9-cis retinoic acid: 0.1, 1, 10 µM; bexarotene 0.1, 1, 10 µM), and Tgf-β (SB431542: 3, 10 µM) pathways were tested for stage- and dose-dependent effects on in vitro anterior-posterior patterning. Our results suggest that with further development, the inclusion of anterior-posterior reporter expression could be part of a battery of high-throughput tests used to identify and characterize teratogens.

中文翻译：

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一种新颖的双重报告基因胚胎干细胞系，用于毒理学诱发的心脏前后格局的扰动毒理学评估。

评估药物和化学物质发育毒性的可靠体外模型将导致胎儿安全性的提高和药物开发成本的降低。经过验证的胚胎干细胞测试（EST）使用小鼠胚胎干细胞（mESCs）的心脏分化来预测体内发育毒性，但未考虑祖细胞在前（心室）和后（心房）阶段的特定模式）车厢。在这项研究中，我们在前和后心脏启动子的控制下生成了带有荧光报告基因的新型双重报告基因mESC系。在转基因小鼠胚胎的新生区隔中观察到报告基因的表达，并使用mESCs进行分化测定，其中使用Wnt（XAV939：3，10 µM），视黄酸（全反式视黄酸：0.1，1，10 µM；9-顺式视黄酸：0.1、1、10 µM；测试了bexarotene 0.1、1、10 µM）和Tgf-β（SB431542：3、10 µM）途径对体外前后图案的阶段和剂量依赖性影响。我们的结果表明，随着进一步的发展，前后报告基因表达的加入可能是用于鉴定和表征致畸物的一系列高通量测试的一部分。