Bentonites, which have been shown to be good mycotoxin adsorbents, have a wide range of biochemical and industrial uses. Although many in vitro studies have shown the effects of bentonite on a variety of cell types, there is no comparable information regarding its effects on neural crest (NC) stem cells. The NC is a collection of multipotent and oligopotent progenitors endowed with both neural and mesenchymal potentials. NC cells originated at trunk level can become peripheral nervous system cells, melanocytes and smooth muscle cells. For this reason, the NC is considered an ideal model for studying the role of different substances on cell differentiation, survival and growth. We have recently shown that aflatoxin B1 (AFB1) decreases the viability and reduces the population of NC cells. In the present work, we have characterized bentonite extracted from the South of Brazil and investigated its effects on NC differentiation and survival, when challenged with AFB1. The results have highlighted the high content of SiO2, Fe2O3 and K2O present in bentonite, with content of MnO, MgO, CaO, Na2O, TiO2 and P2O5 lower than 1%. These results have also shown that bentonite at 0.6mg/mL is not toxic to NC cells in culture. Furthermore, bentonite increased the viability of NC cells previously treated with AFB1 without affecting cell differentiation. The evidence suggests that AFB1 interacted with bentonite by occupying the interlayer spaces, which might have contributed to a reduction of AFB1 availability and, consequently, a reduction in NC damage and an increase in the total number of cells. Our study contributes to a better understanding of the effects of clay minerals on the NC and may help to create new, more effective pharmacological applications to reduce or eliminate the toxic effects of AFB1.