Epigenetic resetting of human pluripotency

• DOI: 10.1101/146712
• Publication Date: 2017-06-07T05:10:17Z
• Authors: Ge Guo; Ferdinand von Meyenn; Maria Rostovskaya; James Clarke; Sabine Dietmann; Duncan Baker; Anna Sahakyan; Samuel Myers; Paul Bertone; Wolf Reik; Kathrin Plath; Austin Smith
• Keywords: Pluripotent Stem Cells; 0301 basic medicine; reprogramming; Cell Differentiation; differentiation; DNA Methylation; methylome; Stem Cells and Regeneration; Embryo, Mammalian; Flow Cytometry; Real-Time Polymerase Chain Reaction; human embryo; Cell Line; Epigenesis, Genetic; Mice; 03 medical and health sciences; Genes, X-Linked; X Chromosome Inactivation; DNA Transposable Elements; Animals; Humans; pluripotent stem cells; Embryonic Stem Cells; In Situ Hybridization, Fluorescence

SUMMARYMuch attention has focussed on conversion of human pluripotent stem cells (PSC) to a more naive developmental status. Here we provide a method for resetting via transient histone deacetylase inhibition. The protocol is effective across multiple PSC lines and can proceed without karyotype change. Reset cells can be expanded without feeders with a doubling time of around 24 hours. WNT inhibition stabilises the resetting process. The transcriptome of reset cells diverges markedly from primed PSC and shares features with human inner cell mass (ICM). Reset cells activate expression of primate-specific transposable elements. DNA methylation is globally reduced to the level in the ICM but is non-random, with gain of methylation at specific loci. Methylation imprints are mostly lost, however. Reset cells can be re-primed to undergo tri-lineage differentiation and germline specification. In female reset cells, appearance of bi-allelic X-linked gene transcription indicates re-activation of the silenced X chromosome. On re-conversion to primed status, XIST-induced silencing restores monoallelic gene expression. The facile and robust conversion routine with accompanying data resources will enable widespread utilisation, interrogation, and refinement of candidate naïve cells.