Human induced pluripotent stem cells (hiPSC) were used to develop an assay format that may deliver information on teratogenicity of drugs. A human pluripotent stem cell scorecard panel was used to monitor the expression of 96 marker genes that are indicative of the stem cell state or differentiation into the ectoderm, mesoderm and endoderm lineages. We selected a human episomal iPS cell line for the assay based on karyotype stability, initial pluripotency, differentiation capacity and overall gene expression variability. The assay is based on embryoid body formation and was developed to be simply automated. In this proof of concept study, we used eight reference compounds (valproic acid, all-trans-retinoic acid, thalidomide, methotrexate, hydroxyurea, ascorbic acid, penicillin G and ibuprofen) to test the technical performance of the assay (readout stability) in concentration-response and time-course experiments. We also found that each compound affected marker gene expression in a different way. Various forms of data analysis identified 19 out of 96 early developmental genes as potential predictive markers for teratogenicity. Machine-learning models were run to exemplify how the assay will be developed further. The preliminary results from these analyses suggest that the assay could be suitable for the pre-screening of candidate pharmaceutical compounds. The approach presented here points a way towards development of a human cell-based assay that could replace the murine EST currently used to screen for early indications of potential teratogenicity of drug candidates.

人类诱导多能干细胞 (hiPSC) 被用于开发一种可以提供药物致畸性信息的检测形式。人类多能干细胞记分卡面板用于监测 96 种标记基因的表达，这些标记基因指示干细胞状态或分化为外胚层、中胚层和内胚层谱系。我们根据核型稳定性、初始多能性、分化能力和整体基因表达变异性选择了人类游离型 iPS 细胞系进行检测。该测定基于拟胚体的形成，并被开发为简单的自动化。在这项概念验证研究中，我们使用了八种参考化合物（丙戊酸、全反式维甲酸、沙利度胺、甲氨蝶呤、羟基脲、抗坏血酸、青霉素 G 和布洛芬）以测试浓度-响应和时程实验中测定的技术性能（读数稳定性）。我们还发现每种化合物都以不同的方式影响标记基因的表达。各种形式的数据分析将 96 个早期发育基因中的 19 个鉴定为潜在的致畸性预测标记。运行机器学习模型以举例说明如何进一步开发该测定。这些分析的初步结果表明，该测定可能适用于候选药物化合物的预筛选。这里介绍的方法指出了一种开发基于人类细胞的检测方法，该检测方法可以取代目前用于筛选候选药物潜在致畸性早期迹象的鼠类 EST。