Rapid generation of functional and homogeneous excitatory human forebrain neurons using Neurogenin-2 (Ngn2)

Induced neuronal \(iN) cells are versatile tools for modeling neurological disorders and human synapse development, and provide a novel cell-based platform for drug screening and discovery [1][2][3][4][5][6][7][8][9] .In 2013 Zhang, Y. et al. study, we reported that inducible expression of a single basic helix-loop-helix transcription factor Neurogenin-2 \(Ngn2) in human embryonic stem \(ES) and induced pluripotent stem \(iPS) cells generates a homogenous population of excitatory neurons that resemble those of cortical upper layer 2/3 neurons in the brain 3 .Within 3-4 weeks in culture, Ngn2-iN cells display mature molecular, cellular and synaptic properties, which can be readily analyzed using various functional assays.Coupled with genome editing and/or patient iPS cell lines, Ngn2-iN cells provide an excellent experimental system that can harness rapid, reliable, and renewable source of human neurons in a culture dish.Here, we describe a stepwise protocol in generating Ngn2-iN cells from both feeder-free human pluripotent stem cells \ (previously published) and feeder-dependent human pluripotent stem cells \(unpublished).