Fig. 1

Generation and characterization of human spinal cord neural progenitor cells from hPSCs. A Schematic procedure for generating hSCNPCs and hiSCNPCs from hPSCs (H9, DC60-3, DC87-3) with all stages, media and factors. DSi represents dual-SMAD inhibitors SB431542 and Dorsomorphin. B Immunostaining of hNMPs markers SOX2 and Brachyury (T) at day 4. Scale bar, 50 μm. C Quantification of SOX2 and Brachyury (T) double positive cells over DAPI of hNMPs-D4. n = 4 independent experiments. Data are represented as mean ± SD. D Principal component analysis of time course samples during hNMPs differentiation and the previously published samples of NKX1-2-GFP sorted human nmps (Verrier et al. 2018). E Cumulative growth curve of hSCNPCs counts over 40 passages. n = 4 independent experiments. Data are represented as mean ± SD. F Immunostaining of hSCNPCs (top panel exhibited pan-NPC markers and proliferation marker; bottom panel exhibited spinal cord specific NPC markers). Scale bar, 50 μm. G Quantification of results shown in Fig. 1F. n = 5 independent experiments. Data are represented as mean ± SD. H Principal component analysis of samples from hESCs (H9) to hSCNPCs at each time point shown in Fig. 1A. I The heat-map showing three regulon groups in samples of hESCs (H9) to hSCNPCs with listing representative regulon transcription factors (numbers of predicted target genes by SCENIC in the brackets) and enriched GO terms for each regulon group. J Comparative analysis of whole transcriptome of hSCNPCs at different passages to dataset from Allen Brain Atlas of developing human brain and previous published dataset of developing human spinal cord