A defined platform of human peri-gastrulation-like biological fate patterning reveals coordination between Reaction-Diffusion and Positional-Information

How position dependent cell fate acquisition occurs during embryogenesis has been a central question in developmental biology. To study this process, we developed a defined, high-throughput assay using BMP4 to induce peri-gastrulation-like fate patterning in geometrically constrained human pluripotent stem cell colonies. We observed that, upon BMP4 treatment, phosphorylated SMAD1 (pSMAD1) activity in the colonies organized into a radial gradient - an observation mechanistically compliant with a BMP4-NOGGIN Reaction-Diffusion (RD) model. Consequent fate acquisition occurred as a function of both the pSMAD1 signaling strength, and induction time - consistent with the Positional-Information (PI) paradigm. Our findings implicate coordination between RD and PI underlying the peri-gastrulation-like fate patterning. This model not only predicts experimental results of perturbing key parameters like colony size, and BMP4 dose, but also identifies experimental conditions that rescue patterning in colonies of sizes that have been reported to be patterning-reticent, and recapitulate RD-like periodic patterns in large colonies.