Epiblast diversification and blood formation in a human pregastrula

The incipient stage of gastrulation in human, when the primitive streak is about to emerge, represents a critical yet underexplored period. Here we present the high-resolution spatial transcriptomic landscape of a human embryo at Carnegie stage 6 (approximately 13–14 days post-conception), a stage at which primitive streak remains invisible and gastrulation-derived mesodermal/endodermal progenitors are not yet transcriptomically detected. We identified an anterior visceral endoderm-like hypoblast population, as well as a trifurcated developmental trajectory of the epiblast, progressing towards the amnion, primitive streak and node/prechordal plate/notochord (axial mesoderm) at subsequent developmental stages1–3. Furthermore, our findings challenge the existing paradigms by revealing that primitive haematopoiesis, involving three blood lineages, initiates in human yolk sac before gastrulation, earlier than previously recognized2,4–7, and that the first blood cells arise from the extra-embryonic mesoderm with a hypoblast rather than epiblast origin. Notably, we identified two spatial zones, each consisting of molecularly distinct yolk sac endoderm and extra-embryonic mesoderm populations, that respectively facilitated the generation of erythro-megakaryocytic lineages and myeloid precursors. These findings provide insights into the onset of gastrulation and the earliest blood formation in humans, with profound implications for advancing stem cell-derived human embryo models and in vitro blood regeneration. High-resolution spatial transcriptome analysis of a human embryo at Carnegie stage 6 reveals three distinct developmental trajectories from the epiblast towards amnion, primitive streak and axial mesoderm, and detects the initiation of haematopoiesis before gastrulation, originating from hypoblast rather than epiblast.