Modeling CNS disorders using hiPSCs holds great potential for elucidating pathogenic mechanisms and identifying potential targets for drug discovery. However, a major challenge to accurately capturing disease-relevant phenotypes using patient-derived hiPSCs is the variability that arises from the heterogeneity in the cellular subtype and the maturation stage of the differentiated neuronal population. In this study, we report directed differentiation of hPSCs to hippocampal granule neurons using a strategy based on developmental hallmarks. We formulated our differentiation approach to mimic the developmental cues by first using antagonists of the WNT, SHH, TGF-β, and BMP pathways to obtain a neural progenitor population patterned for the dorsal forebrain region that gives rise to the hippocampus. Then, we enriched for PROX1+ dentate granule neurons using Wnt3a, a morphogen produced by the cortical hem to establish the DG and CA subfields along the hippocampal primordial (Lee et al., 2000; Machon et al., 2007).
