Illustrative summary of a stepwise treatment protocol for deriving an otic neuronal lineage from undifferentiated H1, H7, and H9 human embryonic stem cells lines. Developmental stages are shown at the top, above a timeline, and key treatments are shown below, using color-coded bars. Abbreviations: D: days; NNE: nonneuronal ectoderm; PPE: preplacodal ectoderm; ONP: otic neuronal progenitor; BMP4: bone morphogenetic protein 4; SHH: Sonic hedgehog; ATRA: all-trans retinoic acid; EGF: epidermal growth factor; BDNF: brain-derived neurotrophic factor; NT3: neurotrophin 3; IGF-1: insulin-like growth factor 1; FGF2: fibroblast growth factor 2; CDM: chemically-defined medium; NIM: neural induction medium; MACS: magnetic-activated cell sorting; FACS: fluorescence-activated cell sorting; p75: low-affinity neurotrophin receptor (p75NTR). This protocol is adapted from Matsuoka et. al., 2017 and 2018.