Fig. 4

Molecular and cellular homogeneity of NESC-spheres. a Sample correlation (Spearman) of transcriptomes among the NESC1-2D-P20, NESC2-2D-P25, NESC1-3D-AP15, NESC2-3D-AP10, NESC1-3D-BP15, and NESC2-3D-BP10. Two cell lines were analyzed and termed as NESC1 (from hESC1) and NESC2 (from BG02). 2D, two-dimensional culture; 3D, three-dimensional suspension-culture; P, passage. A, the CHbFSB+LIF medium; B, the CHbFSB medium. b The heatmap of genes relative to NESCs, radial glial cells (RGCs), intermediate progenitor cells (IPCs), and neurons. c–g A comprehensive transcriptome analysis of single cells using 10x genomics platform. c Uniform manifold approximation and projection (UMAP) plots of 9769 cells from NESC-spheres (P28) colored by cluster annotation. Dots, individual cells. Red, NESCs; green, IPCs (intermediate progenitor cells); blue, DPCs (differentiating-prone cells). d NESCs are colored according to the expression levels of the indicated markers on the UMAP map (red, high; gray, low). e Violin plots show expression levels and distributions of SOX2 and PAX6 in different three clusters. f, g UMAP plots of cells colored by expression of selected marker genes relative to astrocytes, neurons, and IPCs. The different colors indicate the difference of expression level (green, high; gray, low)