Developmental neurotoxicity (DNT) is a branch of toxicology that examines the effects of chemicals on the developing nervous system. Traditional methods for assessing DNT mainly rely on animal testing, which raises ethical concerns, is time-consuming, and expensive. Consequently, there is a shift towards alternative methods, such as in vitro and in silico approaches, which offer faster and more efficient testing. The overall aim of this thesis was to contribute to the development and integration of alternative methods for DNT assessment, employing both in vitro and in silico techniques. In this work, the human neuroblastoma SH-SY5Y cell line was utilized as a robust, cost-effective, and easy-to-use model for DNT evaluation. Through RNA sequencing and morphological observation, it was determined that the SH-SY5Y cell line can differentiate into a more neuron-like phenotype (Paper I). Additionally, neurite outgrowth and the mRNA expression of genes important for neuronal development were studied by exposing the cells to chemicals known to induce DNT (Paper II). The thesis has also focused on acrylamide, a neurotoxic compound that may also cause DNT. In Paper I, it was found that acrylamide inhibited neuronal differentiation by suppressing neurite outgrowth at non-cytotoxic concentrations. Moreover, acrylamide altered the expression of several genes involved in the retinoic acid and CREB signaling pathways. The hypothesis that acrylamide impairs neuronal differentiation by depleting glutathione, leading to oxidative stress, was tested but not supported in the SH-SY5Y cells (Paper III). In Paper IV, we performed an in vitro to in vivo extrapolation by using a novel physiologically based toxicokinetic (PBTK) model for pregnant women, to assess the biological relevance of the acrylamide concentrations that affected neuronal differentiation of SH-SY5Y cells. The results revealed that doses that humans may be exposed to through food intake, resulted in fetal plasma acrylamide concentrations in the low nanomolar range. At these concentrations, attenuated neuronal differentiation has been observed in the SHSY5Y cells. Additionally, effects seen at micromolar concentrations were considered concerning for fetal health in cases of accidental exposure. In conclusion, human neuroblastoma SH-SY5Y cells can serve as a useful cell model for initial screening in DNT assessment, particularly for studying neuronal differentiation as a key neurodevelopmental process. Furthermore, this thesis suggests that acrylamide may pose a risk to the developing brain, as indicated by its effects on differentiation in SH-SY5Y cells and the extrapolation of in vitro concentrations to in vivo doses, by PBTK modeling. However, to validate these findings, further testing in more complex cell culture models is necessary.