High-throughput micropatterning platform reveals Nodal-dependent bisection of peri-gastrulation-associated versus preneurulation-associated fate patterning

PLoS Biol. 2019 Oct 21;17(10):e3000081. doi: 10.1371/journal.pbio.3000081. eCollection 2019 Oct.

Abstract

In vitro models of postimplantation human development are valuable to the fields of regenerative medicine and developmental biology. Here, we report characterization of a robust in vitro platform that enabled high-content screening of multiple human pluripotent stem cell (hPSC) lines for their ability to undergo peri-gastrulation-like fate patterning upon bone morphogenetic protein 4 (BMP4) treatment of geometrically confined colonies and observed significant heterogeneity in their differentiation propensities along a gastrulation associable and neuralization associable axis. This cell line-associated heterogeneity was found to be attributable to endogenous Nodal expression, with up-regulation of Nodal correlated with expression of a gastrulation-associated gene profile, and Nodal down-regulation correlated with a preneurulation-associated gene profile expression. We harness this knowledge to establish a platform of preneurulation-like fate patterning in geometrically confined hPSC colonies in which fates arise because of a BMPs signalling gradient conveying positional information. Our work identifies a Nodal signalling-dependent switch in peri-gastrulation versus preneurulation-associated fate patterning in hPSC cells, provides a technology to robustly assay hPSC differentiation outcomes, and suggests conserved mechanisms of organized fate specification in differentiating epiblast and ectodermal tissues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomechanical Phenomena
  • Body Patterning / genetics
  • Bone Morphogenetic Protein 4 / genetics
  • Bone Morphogenetic Protein 4 / metabolism
  • Bone Morphogenetic Protein 4 / pharmacology*
  • Cell Culture Techniques
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Lineage / drug effects*
  • Cell Lineage / genetics
  • Gastrulation / drug effects
  • Gastrulation / genetics
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental*
  • Genetic Heterogeneity
  • High-Throughput Screening Assays
  • Humans
  • Models, Biological
  • Neurogenesis / drug effects
  • Neurogenesis / genetics
  • Nodal Protein / genetics*
  • Nodal Protein / metabolism
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / drug effects*
  • Pluripotent Stem Cells / metabolism
  • Signal Transduction
  • Surface Properties

Substances

  • BMP4 protein, human
  • Bone Morphogenetic Protein 4
  • NODAL protein, human
  • Nodal Protein