Abstract
Naive pluripotent embryonic stem cells (ESCs) and embryonic germ cells (EGCs) are derived from the preimplantation epiblast and primordial germ cells (PGCs), respectively. We investigated whether differences exist between ESCs and EGCs, in view of their distinct developmental origins. PGCs are programmed to undergo global DNA demethylation; however, we find that EGCs and ESCs exhibit equivalent global DNA methylation levels. Inhibition of MEK and Gsk3b by 2i conditions leads to pronounced reduction in DNA methylation in both cell types. This is driven by Prdm14 and is associated with downregulation of Dnmt3a and Dnmt3b. However, genomic imprints are maintained in 2i, and we report derivation of EGCs with intact genomic imprints. Collectively, our findings establish that culture in 2i instills a naive pluripotent state with a distinctive epigenetic configuration that parallels molecular features observed in both the preimplantation epiblast and nascent PGCs.
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24 February 2013
In the version of this article initially published online, a bar connecting the two central data sets in Figure 3e was inadvertently included. The error has been corrected for the print, PDF and HTML versions of this article.
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Acknowledgements
We thank members of the Smith, Surani and Hajkova labs for stimulating discussions and C. Mulas for comments on the manuscript. We are also grateful to B. Snijders from the Medical Research Council (MRC) Clinical Sciences Centre Proteomics facility and Agilent Technologies for their support regarding the LC-MS analysis. Affymetrix array hybridization and quality control was carried out by the Genomics Laboratory at the MRC Clinical Sciences Centre. A.S. is supported as an MRC Professor. This work was supported by MRC funding to P.H. (MC A652 5PY70) as well as a Wellcome Trust grant to M.A.S. (RG49135) and MRC funding to A.S. (G1001028).
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The study was conceived of and designed by H.G.L., K.R.M. and P.H. Experiments were performed by H.G.L., K.R.M., A.T., V.E., B.N. and P.H. Bioinformatic analysis was performed by T.C. and K.R.M. Blastocyst injections were performed by W.M., N.G. and J.G.K. provided reagents. A.S. and M.A.S. provided critical feedback. H.G.L., K.R.M. and P.H. wrote the manuscript.
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Leitch, H., McEwen, K., Turp, A. et al. Naive pluripotency is associated with global DNA hypomethylation. Nat Struct Mol Biol 20, 311–316 (2013). https://doi.org/10.1038/nsmb.2510
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DOI: https://doi.org/10.1038/nsmb.2510
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