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

• Published in: PLoS biology 2019-10, Vol.17 (10), p.e3000081
• Main Authors: Tewary, Mukul, Dziedzicka, Dominika, Ostblom, Joel, Prochazka, Laura, Shakiba, Nika, Heydari, Tiam, Aguilar-Hidalgo, Daniel, Woodford, Curtis, Piccinini, Elia, Becerra-Alonso, David, Vickers, Alice, Louis, Blaise, Rahman, Nafees, Danovi, Davide, Geens, Mieke, Watt, Fiona M, Zandstra, Peter W
• Format: Article
• Language: English
• Subjects: Biology and life sciences; Biomechanical Phenomena; Body Patterning - genetics; Bone morphogenetic protein 4; Bone Morphogenetic Protein 4 - genetics; Bone Morphogenetic Protein 4 - metabolism; Bone Morphogenetic Protein 4 - pharmacology; Bone morphogenetic proteins; Cell Culture Techniques; Cell Differentiation - drug effects; Cell Line; Cell Lineage - drug effects; Cell Lineage - genetics; Colonies; Developmental biology; Diagnostic reagents industry; Differentiation; Embryonic development; Gastrulation; Gastrulation - drug effects; Gastrulation - genetics; Gene expression; Gene Expression Profiling; Gene Expression Regulation, Developmental; Genetic aspects; Genetic Heterogeneity; Heterogeneity; High-Throughput Screening Assays; Humans; Micropatterning; Models, Biological; Neurogenesis - drug effects; Neurogenesis - genetics; Nodal Protein - genetics; Nodal Protein - metabolism; Pattern formation; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - drug effects; Pluripotent Stem Cells - metabolism; Regeneration (physiology); Regenerative medicine; Signal Transduction; Signaling; Social Sciences; Stem cells; Surface Properties; Tissue engineering
• ISSN: 1545-7885; 1544-9173; 1545-7885
• DOI: 10.1371/journal.pbio.3000081

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.