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Assessing the Wnt-reactivity of cytonemes of mouse embryonic stem cells using a bioengineering approach

Authors
  • Junyent, Sergi1
  • Reeves, Joshua1
  • Habib, Shukry J.1
  • 1 Centre for Stem Cells and Regenerative Medicine, King’s College London, London SE1 9RT, UK
Type
Published Article
Journal
STAR Protocols
Publisher
Elsevier
Publication Date
Sep 15, 2021
Volume
2
Issue
3
Identifiers
DOI: 10.1016/j.xpro.2021.100813
PMID: 34568840
PMCID: PMC8449058
Source
PubMed Central
Keywords
Disciplines
  • Protocol
License
Unknown

Abstract

These protocols investigate the interaction of cytonemes with localized Wnt. Cell-niche signaling between naïve or primed mouse embryonic stem cells (ESCs) and either Wnt-secreting trophoblast stem cells (TSCs) or Wnt signals reconstituted on microbeads can be scrutinized in vitro . This approach analyzes cytoneme reactivity during Wnt-interaction initiation, Ca2+ transients at Wnt-contacting cytonemes, and subsequent pairing between ESCs and Wnt-sources. This pairing interaction is crucial to synthetic embryogenesis, hence this protocol is effective for in vitro studies of developmental biology.

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