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A defined Oct4 level governs cell state transitions of pluripotency entry and differentiation into all embryonic lineages

Nature Cell Biology volume 15, pages 579–590 (2013)Cite this article

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Abstract

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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Figure 1: An ESC level of Oct4 marks pluripotency acquisition.
Figure 2: Low levels of Oct4 expression sustain self-renewal.
Figure 3: Oct4 expression at an ESC level is required for in vitro differentiation.
Figure 4: Oct4 expression at an ESC level is required for in vivo differentiation.
Figure 5: Oct4 binding is converse to Nanog and is linked to downregulation of naive pluripotency genes.
Figure 6: Oct4-low iPSCs self-renew in the absence of pluripotent culture requisites.
Figure 7: A defined Oct4 level is also required for downregulation of key naive pluripotency genes in ESCs.
Figure 8: A defined Oct4 level controls cell state transitions around pluripotency.

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Change history

  • 07 May 2013

    In the version of this Article originally published online, there were errors in Fig. 1k,l. This has been corrected in the HTML and PDF versions.

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Acknowledgements

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.

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  1. Thorold W. Theunissen

    Present address: Present address: Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA,

Authors and Affiliations

  1. Wellcome Trust—Medical Research Council Cambridge Stem Cell Institute and Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK

    Aliaksandra Radzisheuskaya, Rodrigo L. dos Santos, Thorold W. Theunissen & José C. R. Silva

  2. Wellcome Trust—Medical Research Council Cambridge Stem Cell Institute and Department of Physiology, Development and Neuroscience, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK

    Gloryn Le Bin Chia & Jennifer Nichols

  3. Doctoral Programme in Experimental Biology and Biomedicine, Centre for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal

    Rodrigo L. dos Santos

  4. CIMAR/CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos Bragas, Porto 4050-123, Portugal

    L. Filipe C. Castro

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Contributions

A.R. performed and designed the experiments, analysed the data and wrote the manuscript. R.L.S. and G.L.B.C. performed experiments. T.W.T., L.F.C., A.R. and J.S. designed the study. J.N. analysed data. J.S. supervised the study, designed the experiments, analysed the data, and wrote and approved the manuscript.

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Correspondence to José C. R. Silva.

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The authors declare no competing financial interests.

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Supplementary Table 1

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PB-Oct4 iPSCs−/− differentiate into beating heart cells.

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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