Extended Data Fig. 1: iCoMoNSCs characterization.

a, Phase contrast images from different stages of iCoMoNSC generation. Human fibroblasts (left) were reprogrammed into iPS cell colonies (2nd left), which formed embryoid bodies (middle) and then generated neural rosettes (2nd right). Patches of morphologically distinct colonies migrated out of rosettes and were isolated as clones (right). iCoMoNSC clones were successfully generated from 2 WT iPS cell clones. b, Phase contrast image showing the overall homogeneous morphology and pinwheel growth organization of the iCoMoNSCs (representative image from an early passage - passages were imaged regularly). NSC-marker immunofluorescence of iCoMoNSCs at early passage 6 (c, d) and late passage 17 (e, f) with NES, SOX2 and PLZF (c, e) or ZOI (d, f) - early and late passage NSC marker IF was repeated twice. Violin plots showing the number of genes (g) or UMIs (h) detected in all iCoMoNSCs by scRNA-seq coming from replicates from two independent cell culture dishes. Blue data points are cells retained after quality control, yellow data points are cells that were filtered out. i, UMAP of two integrated replicate samples of iCoMoNSCs clustering into 7 clusters, (j) same UMAP representing cell cycle stages or (k) showing the replicates whose distribution across the clusters is shown in (l). m, UMAP with normalized expression of selected genes across all iCoMoNSCs. Scale bars, (a) from left to right: 100 µm, 500 µm, 500 µm, 500 µm, 50 µm, (b) 150 µm, (c-f) 50 µm.