Mouse embryonic stem cells in reproductive toxicology: establishment of a predictive in vitro test module for developmental neurotoxicity testing
2009
Hayess,K. | Slawik,B. | Visan,A. | Sittner,D. | Spielmann,H. | Luch,A. | Seiler,A.
Alemán. Pluripotent mouse embryonic stem (ES) cells are capable of differentiating into many fully specified and functional cell types including neural cells while recapitulating in vivo developmental programs. Therefore, these cells offer the unique opportunityto study adverse effects on neural cell development in vitro after exposure to toxic compounds. In the context of a complex modular strategy for developmental neurotoxicity testing, we are currently developing a predictive in vitro test module using the mouse ES cell line D3. To this end, we established a robust and fast method for differentiation of D3 cells into neurons, astrocytes or oligodendrocytes suitable for testing of chemicals and other compounds. The protocolwas also adapted to a 96-well plate format in order to facilitate higher throughput. Furthermore, to assess adverse effects on neural cell differentiation and proliferation predictive toxicological endpoints have been established using flow cytometryof neuron-specific as well as glia-specific marker proteins and proliferation assays. Here we present the results obtained with selected compounds of known and characterized developmental neurotoxic potency in vivo as well as negative control substances.Comparative evaluation of dose response profiles revealed significant differences in the sensitivity of undifferentiated ES cells, 3T3 fibroblasts and ES cells differentiating into neurons.Overall our results suggest that the mouse ES cell model provides trustful and reliable results in the detection of developmental neurotoxicants.
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