Patient-derived induced pluripotent stem cells (hiPSCs) hold the potential to model disease mechanisms in vitro [6–9], to screen therapeutic targets and to generate autologous cell populations for cell replacement therapies [10, 11]. Many differentiation protocols have been described to produce neuronal cell cultures from human pluripotent stem cells (hPSCs) or neuroepithelial stem (hNES) cells [12–16]. Several brain-patterning factors such as sonic hedgehog (SHH [17]), retinoic acid (RA [18]), fibroblast growth factors (FGFs [19]), insulin growth factors (IGFs [20]) and Wnts [21] have been used to generate specific neural cell types. Existing procedures generate mixed neural cultures, but lack derivation of pure neuronal cultures with balanced inhibitory and excitatory synaptic activities suitable for single cell analysis [22–24].
