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Development. 2018 Sep 21;145(18). pii: dev166025. doi: 10.1242/dev.166025.

Geometrical confinement controls the asymmetric patterning of brachyury in cultures of pluripotent cells.

Author information

1
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, EH16 4UU, UK guillaume.blin@ed.ac.uk.
2
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, EH16 4UU, UK.
3
Univ. Grenoble-Alpes, CEA, CNRS, INRA, Biosciences and Biotechnology Institute of Grenoble, Laboratoire de Physiologie Cellulaire and Végétale, UMR5168, CytoMorpho Lab, 38054 Grenoble, France.
4
Univ. Paris Diderot, CEA, INSERM, Hôpital Saint Louis, Institut Universitaire d'Hematologie, UMRS1160, CytoMorpho Lab, 75010 Paris, France.
5
INSERM U1251, Université Aix-Marseille, MMG, 13885 Marseille, France.

Abstract

Diffusible signals are known to orchestrate patterning during embryogenesis, yet diffusion is sensitive to noise. The fact that embryogenesis is remarkably robust suggests that additional layers of regulation reinforce patterning. Here, we demonstrate that geometrical confinement orchestrates the spatial organisation of initially randomly positioned subpopulations of spontaneously differentiating mouse embryonic stem cells. We use micropatterning in combination with pharmacological manipulations and quantitative imaging to dissociate the multiple effects of geometry. We show that the positioning of a pre-streak-like population marked by brachyury (T) is decoupled from the size of its population, and that breaking radial symmetry of patterns imposes polarised patterning. We provide evidence for a model in which the overall level of diffusible signals together with the history of the cell culture define the number of T+ cells, whereas geometrical constraints guide patterning in a multi-step process involving a differential response of the cells to multicellular spatial organisation. Our work provides a framework for investigating robustness of patterning and provides insights into how to guide symmetry-breaking events in aggregates of pluripotent cells.

KEYWORDS:

Embryonic; Heterogeneity; Micropatterning; Mouse; Self-organisation; Stem cells

PMID:
30115626
PMCID:
PMC6176930
DOI:
10.1242/dev.166025
[Indexed for MEDLINE]
Free PMC Article

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