The Organization for Economic Co-operation and Development (OECD) test guideline 426 for developmental neurotoxicity (DNT) of industrial/environmental chemicals depends primarily on animal experimentation. This requirement raises various critical issues, such as high cost, long duration, the sacrifice of large numbers of animals, and interspecies differences. This study demonstrates an alternative protocol that is simple, quick, less expensive, and standardized to evaluate DNT of many chemicals using human induced pluripotent stem cells (iPSC) and their differentiation to neural progenitor cells (NPC). Initially, concentration-dependent cytotoxicity of 35 DNT chemicals, including industrial materials, insecticides, and clinical drugs, were compared among iPSC, NPC, and two transformed cells, Cos-7 and HepG2, using tetrazolium dye (MTS)-reducing colorimetric and ATP luciferase assays, and IC₅₀ values were calculated. Next, inhibitory effects of the 14 representative chemicals (mainly insecticides) on iPSC differentiation to NPC were evaluated by measuring altered expression of neural differentiation and undifferentiation marker genes. Results show that both iPSC and NPC were much more sensitive to most DNT chemicals than the transformed cells, and 14 chemicals induced differential patterns of marker gene expression, highlighting the validity and utility of the protocol for evaluation and classification of DNT chemicals and preclinical DNT tests for safety assessment.

经济合作与发展组织 (OECD) 的工业/环境化学品发育神经毒性 (DNT) 测试指南 426 主要取决于动物实验。这一要求提出了各种关键问题，例如成本高、持续时间长、牺牲大量动物以及种间差异。本研究展示了一种替代方案，该方案简单、快速、成本较低且标准化，可使用人类诱导多能干细胞 (iPSC) 及其向神经祖细胞 (NPC) 的分化来评估许多化学品的 DNT。最初，在 iPSC、NPC 和两种转化细胞 Cos-7 和 HepG2 之间比较了 35 种 DNT 化学品（包括工业材料、杀虫剂和临床药物）的浓度依赖性细胞毒性，计算了_{50 个}值。接下来，通过测量神经分化和未分化标记基因的表达改变来评估 14 种代表性化学品（主要是杀虫剂）对 iPSC 向 NPC 分化的抑制作用。结果表明，iPSC 和 NPC 对大多数 DNT 化学物质比转化细胞更敏感，14 种化学物质诱导了标记基因表达的差异模式，突出了该协议对 DNT 化学物质的评估和分类以及临床前 DNT 测试的有效性和实用性用于安全评估。