Centre for Stem Cells and Regenerative Medicine, King's College London, London SE1 9RT, UK.
STAR Protoc. 2021 Sep 15;2(3):100813. doi: 10.1016/j.xpro.2021.100813. eCollection 2021 Sep 17.
These protocols investigate the interaction of cytonemes with localized Wnt. Cell-niche signaling between naive or primed mouse embryonic stem cells (ESCs) and either Wnt-secreting trophoblast stem cells (TSCs) or Wnt signals tethered to microbeads can be scrutinized . This approach analyzes cytoneme reactivity during Wnt-interaction initiation, Ca 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 studies of developmental biology. For complete details on the use and execution of this protocol, please refer to Junyent et al. (2020, 2021a, 2021b).
这些方案研究了纤毛与局部 Wnt 的相互作用。可以仔细研究未成熟或初始状态的小鼠胚胎干细胞 (ESCs) 与分泌 Wnt 的滋养层干细胞 (TSC) 之间，或者与微珠固定的 Wnt 信号之间的细胞 - 巢信号。该方法分析了 Wnt 相互作用起始时纤毛的反应性、与 Wnt 接触的纤毛中的 Ca 瞬变，以及随后 ESCs 与 Wnt 源之间的配对。这种配对相互作用对合成胚胎发生至关重要；因此，该方案可有效用于发育生物学研究。有关该方案的使用和执行的完整详细信息，请参阅 Junyent 等人。（2020 年，2021a，2021b）。
