Title:Characterisation of a Neural Teratogenicity Assay Based on Human ESCs Differentiation Following Exposure to Valproic Acid
Volume: 19
Issue: 35
Author(s): S. Colleoni, C. Galli, J. A. Gaspar, K. Meganathan, S. Jagtap, J. Hescheler, A. Sachinidis and G. Lazzari
Affiliation:
Keywords:
Alternative in vitro test, teratogenicity assay, valproic acid, early neural toxicity, embryonic stem cells, neural development, neural
tube, prenatal developmental toxicity
Abstract: The development of in vitro testing strategies for chemical and drug screening is a priority need in order to protect human
health, to increase safety, to reduce the number of animals required for conventional testing methods and finally to meet the deadlines of
current legislations. The aim of this work was to design an alternative testing method based on human embryonic stem cells for the detection
of prenatal neural toxicity. For this purpose we have created a model based on the generation of neural rosettes, reproducing in vitro
the gastrulation events recapitulating the formation of the neural tube in vivo. To validate the model we have exposed this complex cell
system to increasing concentrations of valproic acid, a known teratogenic agent, to analyse the morphological and molecular changes induced
by the toxicant. Specific assays were applied to discriminate between cytotoxicity and specific neural toxicity. Transcriptomic
analysis was performed with a microarray Affimetrix platform and validated by quantitative real time RT-PCR for the expression of
genes involved in early neural development, neural tube formation and neural cells migration, key biological processes in which the effect
of valproic acid is most relevant. The results demonstrated that neural rosette cells respond to valproic acid exposure with molecular
and morphological changes similar to those observed in vivo, indicating that this method represents a promising alternative test for the
detection of human prenatal neural toxicity.
