Microgel culture and spatial identity mapping elucidate the signalling requirements for primate epiblast and amnion formation

Funder: Wellcome Trust; Id: http://dx.doi.org/10.13039/100010269
Funder: Royal Society; Id: http://dx.doi.org/10.13039/501100000288
Funder: AstraZeneca; Id: http://dx.doi.org/10.13039/100004325
Funder: University of Cambridge; Id: http://dx.doi.org/10.13039/501100000735
The early specification and rapid growth of extraembryonic membranes are distinctive hallmarks of primate embryogenesis. These complex tasks are resolved through an intricate combination of signals controlling the induction of extraembryonic lineages and, at the same time, safeguarding the pluripotent epiblast. Here, we delineate the signals orchestrating primate epiblast and amnion identity. We encapsulated marmoset pluripotent stem cells into agarose microgels and identified culture conditions for the development of epiblast- and amnion-spheroids. Spatial identity mapping authenticated spheroids generated in vitro by comparison to marmoset embryos in vivo. We leveraged the microgel system to functionally interrogate the signalling environment of the postimplantation primate embryo. Single-cell profiling of the resulting spheroids demonstrated that ACTIVIN/NODAL signalling is required for embryonic lineage identity. BMP4 promoted amnion formation and maturation, which was counteracted by FGF-signalling. Our combination of microgel culture, single-cell profiling and spatial identity mapping provides a powerful approach to decipher the essential cues for embryonic and extraembryonic lineage formation in primate embryogenesis.