•Bisphenol A and para-nonylphenol affect human placenta development at concentrations detected in human tissues and fluids.•BPA and p-NP decreased trophoblast cell migration and invasion and trophoblast/endothelial interaction.•BPA and p-NP induced differentiation of trophoblast into placental bed giant cells.•BPA and p-NP act on trophoblast cells in a dose-dependent manner. Bisphenol A (BPA) and para-Nonylphenol (p-NP) are chemicals of industrial origin which may influence human reproductive health. The effects of these substances in the prenatal life is an important topic that is receiving greater attention in the developed countries. In this study, human trophoblast cells HTR-8/SVneo were exposed to BPA and p-NP (1 × 10−15, 1 × 10−13, 1 × 10−11, 1 × 10−9 and 1 × 10−7 M) and incubated for 24, 48 and/or 72 h then, examined for the main physiological processes which characterize the extravillous trophoblast. Cell proliferation showed no changes while the processes of cell migration and invasion were both reduced by BPA and p-NP. For each chemical, the activity was higher at lower concentrations with a maximum activity between 1 × 10−13 and 1 × 10−11 M (p < 0.05 for 1 × 10−9 and p < 0.001 for 1 × 10−11 M). Co-culture studies with human umbilical cord endothelial cells (HUVEC) revealed that trophoblast/endothelial interaction was significantly reduced by p-NP at 1 × 10−11 M. Moreover, both chemicals were inducing differentiation of HTR-8/SVneo toward polyploidy by the process of endoreduplication. The estrogen-receptor antagonist ICI significantly reduced p-NP action, while it had no effect on BPA treated cells. In conclusion, p-NP and BPA act on trophoblast cells altering key physiological processes in placenta development. The exact mechanism of action of the chemicals in human trophoblast still needs to be clarified.