There is an increase in effort and interest around the world for developing appropriate disease models for Alzheimer’s disease (AD). Despite such awareness, most of the in vitro AD models do not resemble the AD patient brain environment. To address this critical issue, we treated human embryonic stem cells (hESCs)-derived neurons with monomeric Aβ (1-42) at pathophysiologically more relevant concentrations for longer period compared to other conventional AD studies. To generate forebrain neurons we used a combination of small molecules for the first 9 days of differentiation. The small molecules include inhibitors of SMAD signalling and a Wnt signalling pathway inhibitor to derive central nervous system lineages, followed by other small molecules to derive forebrain neurons. The Aβ (1-42) monomer treatment initiated on day 50 of differentiation for 15 days and the monomer concentrations ranged from 0 -10 μM. Immunocytochemistry demonstrated that there is a reduction in both thickness and length of axons with low concentrations of Aβ (1-42) monomer. Currently, inflammatory cytokine level changes during the Aβ treatment are under investigation by using cytometric bead array (CBA) assay. Overall, the combination of current differentiation protocol and extended Aβ (1-42) treatment successfully produces a more disease relevant model for AD.