Human ES cell-derived neural rosettes reveal a functionally distinct early neural stem cell stage

Abstract

Neural stem cells (NSCs) yield both neuronal and glial progeny, but their differentiation potential toward multiple region-specific neuron types remains remarkably poor. In contrast, embryonic stem cell (ESC) progeny readily yield region-specific neuronal fates in response to appropriate developmental signals. Here we demonstrate prospective and clonal isolation of neural rosette cells (termed R-NSCs), a novel NSC type with broad differentiation potential toward CNS and PNS fates and capable of in vivo engraftment. R-NSCs can be derived from human and mouse ESCs or from neural plate stage embryos. While R-NSCs express markers classically associated with NSC fate, we identified a set of genes that specifically mark the R-NSC state. Maintenance of R-NSCs is promoted by activation of SHH and Notch pathways. In the absence of these signals, R-NSCs rapidly lose rosette organization and progress to a more restricted NSC stage. We propose that R-NSCs represent the first characterized NSC stage capable of responding to patterning cues that direct differentiation toward region-specific neuronal fates. In addition, the R-NSC-specific genetic markers presented here offer new tools for harnessing the differentiation potential of human ESCs.[Keywords: Human embryonic stem cells; neural patterning; neural stem cells; neuronal specification] Supplemental material is available at http://www.genesdev.org.

Retrograde interferon‐gamma signaling induces major histocompatibility class I expression in human‐induced pluripotent stem cell‐derived neurons

“…It has been speculated that part of the success of neurosphere cultures is due to the fact that the 3D architecture more closely recapitulates the in vivo microenvironment of the NSC niche 52, 53. Recently, techniques have been established for reliably generating monolayers of induced pluripotent stem cell (iPSC)‐derived NSCs using supplementation with basic fibroblast growth factor (FGFb) and epidermal growth factor (EGF) 54, 55, 56. However, such cultured monolayer NSCs are more transcriptionally homogeneous than in vivo NSCs, and it has been speculated that this lack of heterogeneity is responsible, at least in part, for the challenges of promoting the differentiation of cultured NSCs into diverse neuronal subtypes 56, 57, 58, 59…”

The Apical Polarity Determinant Crumbs 2 Is a Novel Regulator of ESC-Derived Neural Progenitors

“…ESCs can be efficiently differentiated into neural lineages in adherent monolayer cultures in the absence of serum [20], which closely mimics embryonic neural development [21][22][23]. Cell polarity is also maintained in vitro, as neural progenitors tend to organize themselves into neural rosettes [23][24][25][26]. The formation of neural rosettes is initiated by the acquisition of cellular polarity and marked by apical localization of ZO-1 [27], the neural stem cell marker Prominin1 [28], and members of the Par complex, including Par3 and aPKC [23].…”

“…The latter have been proposed to act as fate determinants in embryonic neural stem cells [29]. Neural rosettes are conserved among species [23,26,30,31] and positive for early neural markers, such as Sox1, Sox2, Nestin, and Pax6 [23,26,31]. Recently, it has been shown in human ESCs that neural rosettes represent an early neural stem cell stage [26].…”

Direct lineage conversion: induced neuronal cells and induced neural stem cells

“…Kim et al (2011a) investigated whether iNS cells could be generated from somatic cells by reprogramming cells with the same four reprogramming factors used to generate iPS cells followed by culture in neural stem (NS) cells medium. They successfully reprogrammed fibroblasts to iNS cells over an abbreviated induction period, and these iNS cells expressed several NS cell-specific markers, including promyelocytic leukemia zinc finger (Plzf), a rosette NSC marker (Elkabetz et al, 2008), and Pax6, an early neural transcription factor (Walther and Gruss, 1991). Various mature neuronal and glial markers were also detected in those induced NS cells after the spontaneous differentiation of cells from isolated colonies.…”