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.
No
PLoS Biol
Article
Científica
7.076
4.488
01/10/2019
000505712400001
2-s2.0-85074378026
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.
bone morphogenetic protein 4; morphogen; protein Nodal; Article; cell differentiation; controlled study; correlational study; down regulation; embryo; embryo pattern formation; gastrulation; genetic association; genetic fingerprinting; geometry; human; human cell; neurulation; pluripotent stem cell line; protein expression; upregulation