Multipotent Caudal Neural Progenitors Derived from Human Pluripotent Stem Cells That Give Rise to Lineages of the Central and Peripheral Nervous System

Abstract

The caudal neural plate is a distinct region of the embryo that gives rise to major progenitor lineages of the developing central and peripheral nervous system, including neural crest and floor plate cells. We show that dual inhibition of the glycogen synthase kinase 3β and activin/nodal pathways by small molecules differentiate human pluripotent stem cells (hPSCs) directly into a preneuroepithelial progenitor population we named “caudal neural progenitors” (CNPs). CNPs coexpress caudal neural plate and mesoderm markers, and, share high similarities to embryonic caudal neural plate cells in their lineage differentiation potential. Exposure of CNPs to BMP2/4, sonic hedgehog, or FGF2 signaling efficiently directs their fate to neural crest/roof plate cells, floor plate cells, and caudally specified neuroepithelial cells, respectively. Neural crest derived from CNPs differentiated to neural crest derivatives and demonstrated extensive migratory properties in vivo. Importantly, we also determined the key extrinsic factors specifying CNPs from human embryonic stem cell include FGF8, canonical WNT, and IGF1. Our studies are the first to identify a multipotent neural progenitor derived from hPSCs, that is the precursor for major neural lineages of the embryonic caudal neural tube. Stem Cells 2015;33:1759–1770

WNT/β-catenin signaling mediates human neural crest induction via a pre-neural border intermediate

“…The absolute requirement of WNT activity in hESC renewal is contested but it appears that nuclear β-catenin signaling is dispensable for ESC maintenance (Kim et al, 2013) and WNT activation leads to differentiation of hESCs (Davidson et al, 2012;Dravid et al, 2005). Combining various chemically defined conditions, cell plating strategies, and activation of signaling pathways, such as activin, Nodal and FGF, WNT activation via GSK3 inhibition or constitutively active β-catenin can direct the differentiation of hESCs into a number of embryonic cell types, including primitive streak/mesendoderm (Funa et al, 2015;Gouti et al, 2014;Nakanishi et al, 2009;Sumi et al, 2008;Tsakiridis et al, 2015), paraxial mesoderm (Shelton et al, 2014;Tan et al, 2013;Umeda et al, 2012), lateral plate mesoderm (Tan et al, 2013), caudal neural plate, and axial stem cell progenitors (Denham et al, 2015;Tsakiridis et al, 2015). ) and neural border genes (NB: PAX7, PAX3, MSX1, TFAP2A) genes to WNT, BMP and FGF signaling analyzed by RT-PCR in D3+CHIR cultures exposed to specific modulators.…”

White paper on guidelines concerning enteric nervous system stem cell therapy for enteric neuropathies

“…ES and iPS cells can be manipulated in vitro to induce neural crest-like phenotype (Chambers et al, 2009; Denham et al, 2015; Hotta et al, 2009; Lee et al, 2010, 2007) and the subsequent engraftment or coculture with embryonic gut tissue (chick or mouse) can induce enteric neuron-like cells (Denham et al, 2015; Hotta et al, 2009; Sasselli et al, 2012a). Recently, as mentioned above, protocols to obtain large numbers of ENS neural progenitors, that colonised the gut of Ednrb mutant mice following transplantation, have been published (Fattahi et al, 2016).…”

Chemotherapy-Induced Neuropathy and Drug Discovery Platform Using Human Sensory Neurons Converted Directly from Adult Peripheral Blood

“…Culture conditions developed by our lab reported here (Table , condition 3) were found to be optimal for expansion, yielding 25× greater number of iNPCs after eight passages (Fig. F) than the other two methods and formats for NPC expansion we tested (Table , condition 1 and 2) . In all conditions, cells retained their PAX6, Nestin, and SOX2 expression indicating that the cultures are broadly stable and not prone to differentiation (Supporting Information Fig.…”