Establishment of a human cell-based in vitro battery to assess developmental neurotoxicity hazard of chemicals

Blum Jonathan, Masjosthusmann Stefan, Bartmann Kristina, Bendt Farina, Dolde Xenia, Dönmez Arif, Förster Nils, Holzer Anna-Katharina, Hübenthal Ulrike, Keßel Hagen Eike, Kilic Sadiye, Klose Jördis, Pahl Melanie, Stürzl Lynn-Christin, Mangas Iris, Terron Andrea, Crofton Kevin M., Scholze Martin, Mosig Axel, Leist Marcel, Fritsche Ellen
DOI: 10.1016/j.chemosphere.2022.137035
PMID: 36328314
Keyword: Brain development · DNT · In vitro testing · Stem cell · Testing battery


Developmental neurotoxicity (DNT) is a major safety concern for all chemicals of the human exposome. However, DNT data from animal studies are available for only a small percentage of manufactured compounds. Test methods with a higher throughput than current regulatory guideline methods, and with improved human relevance are urgently needed. We therefore explored the feasibility of DNT hazard assessment based on new approach methods (NAMs). An in vitro battery (IVB) was assembled from ten individual NAMs that had been developed during the past years to probe effects of chemicals on various fundamental neurodevelopmental processes. All assays used human neural cells at different developmental stages. This allowed us to assess disturbances of: (i) proliferation of neural progenitor cells (NPC); (ii) migration of neural crest cells, radial glia cells, neurons and oligodendrocytes; (iii) differentiation of NPC into neurons and oligodendrocytes; and (iv) neurite outgrowth of peripheral and central neurons. In parallel, cytotoxicity measures were obtained. The feasibility of concentration-dependent screening and of a reliable biostatistical processing of the complex multi-dimensional data was explored with a set of 120 test compounds, containing subsets of pre-defined positive and negative DNT compounds. The battery provided alerts (hit or borderline) for 24 of 28 known toxicants (82% sensitivity), and for none of the 17 negative controls. Based on the results from this screen project, strategies were developed on how IVB data may be used in the context of risk assessment scenarios employing integrated approaches for testing and assessment (IATA).