Oct4 is considered a master transcription factor for pluripotent cell self-renewal, but its biology remains poorly understood. Here, we investigated the role of Oct4 using the process of induced pluripotency. We found that a defined embryonic stem cell (ESC) level of Oct4 is required for pluripotency entry. However, once pluripotency is established, the Oct4 level can be decreased up to sevenfold without loss of self-renewal. Unexpectedly, cells constitutively expressing Oct4 at an ESC level robustly differentiated into all embryonic lineages and germline. In contrast, cells with low Oct4 levels were deficient in differentiation, exhibiting expression of naive pluripotency genes in the absence of pluripotency culture requisites. The restoration of Oct4 expression to an ESC level rescued the ability of these to restrict naive pluripotent gene expression and to differentiate. In conclusion, a defined Oct4 level controls the establishment of naive pluripotency as well as commitment to all embryonic lineages.
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We thank W. Mansfield and C-E. Dumeau for blastocyst injections and morula aggregations, R. Walker for flow cytometry, and M. McLeish and H. Skelton for histological processing of teratomas. We are grateful to H. Niwa for providing mice with different Oct4 genotypes and A. Smith and J. Betschinger for providing plasmids. We are also grateful to Y. Costa and P. Shliaha for technical assistance and H. Stuart for critical reading of the manuscript. The study was supported by Wellcome Trust Fellowship WT086692MA. J.C.R.S. is a Wellcome Trust Career Development Fellow. A.R. is a recipient of the Darwin Trust of Edinburgh Postgraduate Scholarship.
The authors declare no competing financial interests.
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Embryoid body outgrowths of PB-Oct4 iPSCs−/− contain beating heart cells 3 days after plating on gelatine-coated dishes. (AVI 4310 kb)
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Radzisheuskaya, A., Le Bin Chia, G., dos Santos, R. et al. A defined Oct4 level governs cell state transitions of pluripotency entry and differentiation into all embryonic lineages. Nat Cell Biol 15, 579–590 (2013). https://doi.org/10.1038/ncb2742
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