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

Shih P-Y, Kreir M, Kumar D, Seibt F, Pestana F, Schmid B, Holst B, Clausen C, Steeg R, Fischer B, Pita-Almenar J, Ebneth A, and Cabrera-Socorro A.

Stem Cell Research


Using two gene-edited hiPSC lines with a doxycycline-inducible cassette driving NGN2 or NGN2-T2A-GFP expression the authors demonstrate a method for differentiating neuronal cells amenable to scale-up and screening. After 1 week the cells express structural neuronal markers. Function was characterized on MEA and patch-clamp using hiPSC-derived and primary astrocyte coculture. Synchronous network activity was observed on the MEA and activity was comparable to primary mouse neurons.