ABSTRACT The shift from collective migration to differentiation is a crucial process in epithelial biology but recreating this intricate transition has thus far proved elusive. We provide experimental, mechanistic, in vivo , and bioinformatic data supporting an undoubtable link between human pluripotent stem cell (hPSC)- derived collectively migrating hepatoblasts (MHB), and transcriptionally mature, functional hPSC- hepatocytes (HEPs), which incorporates two unrecognized steps. The protocol induces FOXA-dependent induction of HBs, leading to TBX3-positive, YAP-TEAD active MHB’s which provide a transcriptional match with murine liver E9.5 MHBs. Simple cultivation changes trigger MHB’s to rapidly form functional day 18 HEPs, predicted by a deep-learning designed gene circuit, resulting in a ∼236% fold- increase in maturation (PACNet), on par with the highest score, but with enhanced global transcriptional shaping. Overall, incorporating the MHB to HEP transition establishes a new, unrecognized, and highly efficient mechanism for differentiation that can be cumulatively integrated with existing methods to overcome barriers to maturation.

Linking collective migration/growth to differentiation boosts global shaping of the transcriptome and exhibits a grasshopper effect for driving maturation