title: Wnt/β-catenin and FGF signalling direct the specification and maintenance of a neuromesodermal axial progenitor in ensembles of mouse embryonic stem cells.
abstract: The development of the central nervous system is known to result from two sequential events. First, an inductive event of the mesoderm on the overlying ectoderm that generates a neural plate that, after rolling into a neural tube, acts as the main source of neural progenitors. Second, the axial regionalization of the neural plate that will result in the specification of neurons with different anteroposterior identities. Although this description of the process applies with ease to amphibians and fish, it is more difficult to confirm in amniote embryos. Here, a specialized population of cells emerges at the end of gastrulation that, under the influence of Wnt and FGF signalling, expands and generates the spinal cord and the paraxial mesoderm. This population is known as the long-term neuromesodermal precursor (NMp). Here, we show that controlled increases of Wnt/β-catenin and FGF signalling during adherent culture differentiation of mouse embryonic stem cells (mESCs) generates a population with many of the properties of the NMp. A single-cell analysis of gene expression within this population reveals signatures that are characteristic of stem cell populations. Furthermore, when this activation is triggered in three-dimensional aggregates of mESCs, the population self-organizes macroscopically and undergoes growth and axial elongation that mimics some of the features of the embryonic spinal cord and paraxial mesoderm. We use both adherent and three-dimensional cultures of mESCs to probe the establishment and maintenance of NMps and their differentiation.
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Turner
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: Penelope C
authors.initials: PC
authors.lastname: Hayward
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Baillie-Johnson
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: Pau
authors.initials: P
authors.lastname: Rué
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Broome
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: Fernando
authors.initials: F
authors.lastname: Faunes
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK ama11@gen.cam.ac.uk.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Martinez Arias
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D019070:Cell Lineage / Q000502:physiology*; D002490:Central Nervous System / Q000196:embryology*; D004627:Embryonic Induction / Q000502:physiology*; D005346:Fibroblast Growth Factors / Q000502:physiology*; D005434:Flow Cytometry; D005453:Fluorescence; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D008648:Mesoderm / Q000196:embryology*; D051379:Mice; D009024:Morphogenesis / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D059010:Single-Cell Analysis; D059008:Time-Lapse Imaging; D060449:Wnt Signaling Pathway / Q000502:physiology*
chemical_list: D005346:Fibroblast Growth Factors
keywords: Mesodermal; Morphogenesis; Neural; Stem cells; Wnt signalling