The derivation and maintenance of hPSC in stable naïve pluripotent states has wide impact in human developmental biology. However, hPSC are unstable in classical naïve 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 naïve 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 naïve-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, naïve reversion of a broad repertoire of conventional hiPSC reduced lineage-primed gene expression, and significantly improved their multi-lineage differentiation capacities. Stable naïve hPSC with reduced genetic variability and improved functional pluripotency will have great utility in regenerative medicine and human disease modeling.
- Received June 6, 2016.
- Accepted September 11, 2016.
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