Rostrocaudal patterning and neural crest differentiation of human pre-neural spinal cord progenitors in vitro

The spinal cord emerges from a niche of neuromesodermal progenitors (NMPs) formed and maintained by WNT/fibroblast growth factor (FGF) signals at the posterior end of the embryo. NMPs can be generated from human pluripotent stem cells and hold promise for spinal cord replacement therapies. However, NMPs are transient, which compromises production of the full range of rostrocaudal spinal cord identities in vitro. Here we report the generation of NMP-derived pre-neural progenitors (PNPs) with stem cell-like self-renewal capacity. PNPs maintain pre-spinal cord identity for 7–10 passages, dividing to self-renew and to make neural crest progenitors, while gradually adopting a more posterior identity by activating colinear HOX gene expression. The HOX clock can be halted through GDF11-mediated signal inhibition to produce a PNP and NC population with a thoracic identity that can be maintained for up to 30 passages. [Display omitted] •NMPs give rise to PNPs that undergo full, rostrocaudal diversification over time (7–10 passages)•PNPs self-renew and can differentiate to SOX10+ NC with corresponding rostrocaudal identity•GDF signaling regulates the upregulation of sacral HOX genes in PNPs/NC•TGF-β signaling inhibition allows long-term stabilization of PNPs/NC in a thoracic identity In this article, Cooper and colleagues describe the generation and characterization of human pre-neural progenitors (PNPs) that undergo full rostrocaudal diversification, demonstrate stem cell self-renewing characteristics, and differentiate to SOX10+ neural crest (NC) progenitors of corresponding rostrocaudal identity. PNPs and NC progenitors can be maintained, for up to 30 passages, in a thoracic identity by inhibiting GDF11-mediated sacral HOX gene expression.