Development of a fully human assay combining NGN2-inducible neurons co-cultured with iPSC-derived astrocytes amenable for electrophysiological studies

Neurogenin 2 encodes a neural-specific transcription factor (NGN2) able to drive neuronal fate on somatic and stem cells. NGN2 is expressed in neural progenitors within the developing central peripheral nervous systems. Overexpression of human induced pluripotent cells (hiPSCs) or embryonic has been shown efficiently trigger conversion neurons. Here we describe two gene-edited hiPSC lines harbouring doxycycline (DOX)-inducible cassette AAVS1 locus driving expression (BIONi010-C-13) NGN2-T2A-GFP (BIONi010-C-15). By introducing NGN2-expressing cassette, reduce variability associated with conventional over-expression methods such as viral transduction, making these amenable for scale-up production screening processes. DOX-treated hiPSCs convert phenotype one week display structural markers Beta-III tubulin tau. We performed functional characterization NGN2-neurons co-cultured hiPSC-derived astrocytes “fully-humanized” set up. Passive properties were indistinguishable from mouse primary while displaying variable activity extracellular recordings multi-electrode arrays (MEAs). demonstrate that neurons can be comparable gold standard used electrophysiology. Both are globally available via EBiSC repository at https://cells.ebisc.org/.