Axial progenitors generate trunk neural crest cells in vitro

The in vitro generation of neural crest (NC) cells from human pluripotent stem cells (hPSCs) is a valuable approach to study human NC biology and isolate NC derivatives for disease modelling/regenerative medicine applications. However, conventional differentiation protocols induce only a modest yield of NC cells corresponding to the trunk level. Here we show that trunk NC cells and, their downstream derivatives, sympathoadrenal progenitors, can be produced at a high efficiency from hPSC-derived axial progenitors, the in vitro counterparts of the posteriorly-located drivers of embryonic axis elongation. Moreover, using transcriptome analysis, we define the molecular signatures associated with the emergence of human NC cells of distinct axial identities. Collectively, our findings indicate that a post-cranial NC state is achieved through two different routes: the birth of cardiac and vagal NC is facilitated by retinoic acid-induced posteriorisation of an anterior precursor whereas a trunk fate relies on a posterior axial progenitor intermediate. Gene expression profiling utilised total RNA extracted from ES cells (N=3); hPSC derived Cranial neural crest precursors (N=3); hPSC derived Cranial neural crest cells (N=3); hPSC derived Cranial neural crest cells after RA treatment to posteriorise (N=3); hPSC derived Neuromesodermal progenitors (N=3); hPSC derived Trunk neural crest progenitors (N=3); hPSC derived trunk neural crest cells (N=3)