The advantage of using this scalable platform is that it should be possible to induce Ngn2-neurons from independent NPC lines derived from several hiPSCs from many individuals in parallel. To test this, we first evaluated the efficacy of CMV-rtTA and TetO-hNGN2-P2A-eGFP-T2A-PuroR lentiviral transduction across ten NPC lines (three NPC lines each from two controls: NSB553-1-1, NSB553-2-3, NSB553-3-C; NSB2607-1-4, NSB2607-3-1, NSB2607-4-1), and one NPC line each from a third control (NSB690-1-4) as well as from three schizophrenia patients (NSB581-1-2, NSB1442-4-3, NSB2513-1-4). Even without puromycin selection, we robustly achieved >80%-GFP positive cells by florescent activated cell sorting (Fig. 1E). Although minor variations between NPC lines were observed, lentiviral transduction efficiency was not diagnosis-dependent, showing at least as much intra-individual variation as inter-individual variation.
