Fate-Patterning of 2D Gastruloids and Ectodermal Colonies Using Micropatterned Human Pluripotent Stem Cells

Methods Mol Biol. 2021:2258:119-130. doi: 10.1007/978-1-0716-1174-6_9.

Abstract

In the developing mammalian embryo, intercellular signaling allows cells to self-organize to create spatial patterns of different cell fates. This process is challenging to study because of the difficulty of observing or manipulating embryos on the spatial and temporal scales required. In vitro models can provide a complement to in vivo systems for addressing these issues. These models are also the only windows we have into early human development. Here we provide protocols for two systems based on differentiating human pluripotent stem cells in micropatterned colonies on defined size and shape. The first model replicates the patterning of the germ layers at gastrulation, while the second replicates the medial-lateral patterning of the ectoderm. These systems allow study of how signaling underlies self-organized patterning at stages of development which are otherwise inaccessible.

Keywords: Cellular communication; Ectoderm; Gastruloids; Human embryonic stem cells (hESCs); Micropatterns; Self-organization; Signaling; Tissue patterning.

MeSH terms

  • Cell Communication
  • Cell Differentiation*
  • Cell Lineage*
  • Cell Shape
  • Cell Size
  • Cells, Cultured
  • Ectoderm / cytology
  • Ectoderm / physiology*
  • Fluorescent Antibody Technique
  • Gastrulation*
  • Gene Expression Regulation, Developmental
  • Human Embryonic Stem Cells / physiology*
  • Humans
  • Microscopy, Fluorescence
  • Signal Transduction
  • Time Factors