Tankyrase inhibition promotes a stable human naïve pluripotent state with improved functionality

The derivation and maintenance of hPSC in stable naive pluripotent states has wide impact in human developmental biology. However, hPSC are unstable in classical naive mouse ESC WNT and MEK/ERK signal inhibition (2i) culture. We show that a broad repertoire of conventional human embryonic stem cell (hESC) and transgene-independent hiPSC lines could be reverted to stable human preimplantation ICM-like naive states with only WNT, MEK/ERK, and tankyrase inhibition (LIF-3i). LIF-3i-reverted hPSC retained normal karyotypes and attained defining mouse ESC-like functional features including high clonal self-renewal, independence from MEK-ERK signalling, dependence on JAK-STAT3 and BMP4 signaling, and naive-specific transcriptional and epigenetic configurations. Tankyrase inhibition promoted a stable acquisition of a human preimplantation ICM-like ground state via modulation of WNT signalling, and was most efficacious in efficiently reprogrammed conventional hiPSC. Importantly, naive reversion of a broad repertoire of conventional hiPSC reduced lineage-primed gene expression, and significantly improved their multi-lineage differentiation capacities. Stable naive hPSC with reduced genetic variability and improved functional pluripotency will have great utility in regenerative medicine and human disease modeling.