Multimodal characterization of murine gastruloid development

Suppinger, Simon; Zinner, Marietta; Aizarani, Nadim; Lukonin, Ilya; Ortiz, Raphaël; Azzi, Chiara; Stadler, Michael; Vianello, Stefano; Palla, Giovanni; Kohler, Hubertus; Mayran, Alexandre; Liberali, Prisca

doi:10.1016/j.stem.2023.04.018

Cited by 24 publications

(25 citation statements)

References 99 publications

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“…For some time-points, cells were present in both clusters, indicating that the temporal resolution adequately documents the major transitions. This also indicated that, after Chiron treatment, gastruloids are not always perfectly synchronised, as previously reported 14 .…”

Section: Timing Of Cell Fate Transitionsupporting

confidence: 86%

“…This self-organization capacity may involve distinct cell signalling, mechanical constraints between inner and outer cells, or a differential accessibility to Chiron, as well as heterogeneity in the starting cellular state. To limit this latter point, gastruloids were generated from ESCs grown in a medium containing GSK3i and MAPKi (referred to as 2i) where ESCs cultures is more homogeneous 11,14,16,24 (Fig. S1A, B).…”

Section: Gastruloid Self-organization Is Resilient To Intrinsic and E...mentioning

confidence: 99%

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Cadherins modulate the self-organizing potential of gastruloids

Mayran,

Kolly,

Lopez-Delisle

et al. 2023

Preprint

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Gastruloids have recently emerged as an efficient four-dimensional model for studying some aspects of post-implantation embryonic patterning. They undergo gastrulation-like processes leading to the self-organization into highly reproducible biological objects. Here, we sought to uncover the molecular and cellular mechanism underlying this remarkable property. We report that self-organization competence is associated with a cell-specific coordination of a Cadherin switch. We find that N-Cadherin hinders gastruloids morphogenetic competence, for its inactivation leads to the formation of trunk-like structures in absence of extra-cellular matrix analogues. In contrast,E-Cadherinrepression bySnai1is critical for self-organization:Snai1establishes a cell-specific repressive pace by triggering the repression of a pluripotency-associated transcription program and its chromatin landscape, thus allowing a proper transition from E-to N-Cadherin to occur. Altogether, this work establishes a molecular mechanism that integrates the exit from pluripotency and the pace of cell differentiation, leading to the observed self-organizing potential of gastruloids.

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Section: Timing Of Cell Fate Transitionsupporting

confidence: 86%

Section: Gastruloid Self-organization Is Resilient To Intrinsic and E...mentioning

confidence: 99%

Cadherins modulate the self-organizing potential of gastruloids

Mayran,

Kolly,

Lopez-Delisle

et al. 2023

Preprint

Self Cite

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“…While cells display pronounced elongation during the process, cell speed peaks after Wnt activation. The observed changes in cell shape, coupled with increased motility, suggest at least a partial EMT-like transition in gastruloids [26]. This trend remains consistent between gastruloids embedded in Matrigel and gastruloids grown in suspension (Extended Data Fig.…”

Section: Main Textsupporting

confidence: 76%

“…Gastruloids used in this study have been prepared as previously described [26]. In brief mouse embryonic stem cells (ESC) were maintained on gelatin-coated culture plates (6-well) in N2B27 medium consisting of 50 % DMEM/F12 (Catalog no.…”

Section: Methodsmentioning

confidence: 99%

“…Such insights into cellular behavior and maturation processes would not have been attainable without the combination of live imaging and immunofluorescence within the entire organoid volume. Unlike most other samples we imaged, gastruloids are dense structures aggregated from mouse embryonic stem cells [26] and thus are challenging to image with single cell resolution. Hence, we made use of this model system, to display the resolving capability of our light sheet microscope to achieve single cell resolution and to analyze the motility and shape features of individual cells in developing gastruloids.…”

Section: Main Textmentioning

confidence: 99%

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Open top multi sample dual view light sheet microscope for live imaging of large multicellular systems

Moos,

Suppinger,

de Medeiros

et al. 2023

Preprint

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Multicellular systems grow over the course of weeks from single cells to tissues or even full organisms composed of several thousands of cells, making live imaging of such samples challenging. To bridge these wide spatiotemporal scales, we present an open top dual view and dual illumination light sheet microscope dedicated for live imaging of large specimens at single cell resolution. The novel configuration of objectives together with a flexible and customizable multi-well mounting system combines dual view with high throughput multi-position imaging. We use the newly developed microscope to image a wide variety of samples and highlight its capabilities to gain quantitative single-cell information in large specimens such as mature intestinal organoids and gastruloids.

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Three‐dimensional stem cell models of mammalian gastrulation

Turner,

Martinez Arias

2024

BioEssays

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Gastrulation is a key milestone in the development of an organism. It is a period of cell proliferation and coordinated cellular rearrangement, that creates an outline of the body plan. Our current understanding of mammalian gastrulation has been improved by embryo culture, but there are still many open questions that are difficult to address because of the intrauterine development of the embryos and the low number of specimens. In the case of humans, there are additional difficulties associated with technical and ethical challenges. Over the last few years, pluripotent stem cell models are being developed that have the potential to become useful tools to understand the mammalian gastrulation. Here we review these models with a special emphasis on gastruloids and provide a survey of the methods to produce them robustly, their uses, relationship to embryos, and their prospects as well as their limitations.

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Multimodal characterization of murine gastruloid development

Cited by 24 publications

References 99 publications

Cadherins modulate the self-organizing potential of gastruloids

Cadherins modulate the self-organizing potential of gastruloids

Open top multi sample dual view light sheet microscope for live imaging of large multicellular systems

Three‐dimensional stem cell models of mammalian gastrulation

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