In Vitro Toxicology and Biomedicine, Department inaugurated by the Doerenkamp-Zbinden Foundation, University of Konstanz, Box 657, Universitaetsstr. 10, 78457, Konstanz, Germany.
Research Training Group RTG1331, Konstanz, Germany.
Arch Toxicol. 2017 Nov;91(11):3613-3632. doi: 10.1007/s00204-017-1977-y. Epub 2017 May 5.
Many in vitro tests have been developed to screen for potential neurotoxicity. However, only few cell function-based tests have been used for comparative screening, and thus experience is scarce on how to confirm and evaluate screening hits. We addressed these questions for the neural crest cell migration test (cMINC). After an initial screen, a hit follow-up strategy was devised. A library of 75 compounds plus internal controls (NTP80-list), assembled by the National Toxicology Program of the USA (NTP) was used. It contained some known classes of (developmental) neurotoxic compounds. The primary screen yielded 23 confirmed hits, which comprised ten flame retardants, seven pesticides and six drug-like compounds. Comparison of concentration-response curves for migration and viability showed that all hits were specific. The extent to which migration was inhibited was 25-90%, and two organochlorine pesticides (DDT, heptachlor) were most efficient. In the second part of this study, (1) the cMINC assay was repeated under conditions that prevent proliferation; (2) a transwell migration assay was used as a different type of migration assay; (3) cells were traced to assess cell speed. Some toxicants had largely varying effects between assays, but each hit was confirmed in at least one additional test. This comparative study allows an estimate on how confidently the primary hits from a cell function-based screen can be considered as toxicants disturbing a key neurodevelopmental process. Testing of the NTP80-list in more assays will be highly interesting to assemble a test battery and to build prediction models for developmental toxicity.
许多体外测试已被开发出来以筛选潜在的神经毒性。然而，只有少数基于细胞功能的测试被用于比较筛选，因此在如何确认和评估筛选结果方面经验有限。我们针对神经嵴细胞迁移测试（cMINC）解决了这些问题。在初步筛选后，设计了一个后续策略。美国国家毒理学计划（NTP）汇编的一个包含 75 种化合物和内部对照的库（NTP80 列表）被用于该测试。它包含了一些已知的（发育）神经毒性化合物类别。初步筛选产生了 23 个确认的阳性结果，其中包括十种阻燃剂、七种农药和六种类药物化合物。迁移和活力的浓度反应曲线比较表明，所有阳性结果均具有特异性。迁移抑制的程度为 25-90%，两种有机氯农药（DDT、七氯）最为有效。在本研究的第二部分，（1）在防止增殖的条件下重复 cMINC 测定；（2）使用 Transwell 迁移测定作为不同类型的迁移测定；（3）追踪细胞以评估细胞速度。一些有毒物质在不同的测定中具有很大的差异，但每个阳性结果都在至少一种额外的测试中得到了确认。这项比较研究可以评估基于细胞功能的筛选中的主要阳性结果作为干扰关键神经发育过程的有毒物质的可信度。在更多的测定中测试 NTP80 列表将非常有趣，可以组装一个测试电池并建立发育毒性的预测模型。
