Abstract

Pluripotency, the ability to generate any cell type of the body, is an evanescent attribute of embryonic cells. Transitory pluripotent cells can be captured at different time points during embryogenesis and maintained as embryonic stem cells or epiblast stem cells in culture. Since ontogenesis is a dynamic process in both space and time, it seems counterintuitive that these two temporal states represent the full spectrum of organismal pluripotency. Here we show that by modulating culture parameters, a stem-cell type with unique spatial characteristics and distinct molecular and functional features, designated as region-selective pluripotent stem cells (rsPSCs), can be efficiently obtained from mouse embryos and primate pluripotent stem cells, including humans. The ease of culturing and editing the genome of human rsPSCs offers advantages for regenerative medicine applications. The unique ability of human rsPSCs to generate post-implantation interspecies chimaeric embryos may facilitate our understanding of early human development and evolution.

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Accessions

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray, RNA-seq, ChIP-seq and MethylC-seq data have been deposited in the Gene Expression Omnibus under accession number GSE60605.

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Acknowledgements

We would like to thank S. Mitalipov and J. Thomson for providing rhesus ESCs and iPSCs, respectively, F. Gage for providing chimpanzee iPSCs, K. Zhang for assistance with cell line derivation, M. Ku of the H. A. and Mary K. Chapman Charitable Foundations Genomic Sequencing Core for performing RNA-seq and mouse ChIP-seq experiments, M. Chang of the Integrative Genomic and Bioinformatics Core for bioinformatics analysis, W. T. Berggren and the staff of the Salk STEM Core for preparation of custom-mTeSR1 base medium and supply of validated stem culture materials, G. Pao and K. Hasegawa for discussions, Y. Dayn from transgenic core facility and J. Luo for blastocyst injections, Y. Tsunekawa for providing the mutant eGFP human ESCs reporter line, E. O’Connor and K. Marquez of Human Embryonic Stem Cell Core Facility of Sanford Consortium for Regenerative Medicine for FACS sorting, R. H. Benitez, A. Goebl, R. D. Soligalia for assistance with genome editing, M. F. Pera for critical reading of the manuscript, and M. Schwarz, and P. Schwarz for administrative help. M.L. and K.S. are supported by a California Institute for Regenerative Medicine Training Grant. We thank J. L. Mendoza for his support on this project. This work was funded in part by UCAM (mouse studies). J.R.E. is an Investigator of the Howard Hughes Medical Institute. P.G. was supported by Fundacion Pedro Guillen. Work in the laboratory of J.C.I.B. was supported by G. Harold and Leila Y. Mathers Charitable Foundation, The Leona M. and Harry B. Helmsley Charitable Trust and The Moxie Foundation.

Author information

Author notes

    • Daiji Okamura

    Present address: Department of Advanced Bioscience, Graduate School of Agriculture, Kinki University, 3327-204 Nakamachi, Nara 631-8505, Japan.

    • Jun Wu
    •  & Daiji Okamura

    These authors contributed equally to this work.

Affiliations

  1. The Salk Institute for Biological Studies, Gene Expression Laboratory, La Jolla, California 92037, USA

    • Jun Wu
    • , Daiji Okamura
    • , Mo Li
    • , Keiichiro Suzuki
    • , Li Ma
    • , Zhongwei Li
    • , Isao Tamura
    • , Marie N. Krause
    • , Tomoaki Hishida
    • , Yuta Takahashi
    • , Emi Aizawa
    • , Na Young Kim
    • , Concepcion Rodriguez Esteban
    •  & Juan Carlos Izpisua Belmonte
  2. Howard Hughes Medical Institute, The Salk Institute for Biological Studies, La Jolla, California 92037, USA

    • Chongyuan Luo
    •  & Joseph R. Ecker
  3. The Salk Institute for Biological Studies, Genomic Analysis Laboratory, La Jolla, California 92037, USA

    • Chongyuan Luo
    • , Yupeng He
    • , Joseph R. Nery
    • , Zhuzhu Zhang
    •  & Joseph R. Ecker
  4. The Salk Institute for Biological Studies, Integrated Genomics, La Jolla, California 92037, USA

    • Chris Benner
  5. Ludwig Institute for Cancer Research, University of California, San Diego School of Medicine, Department of Cellular and Molecular Medicine, 9500 Gilman Drive, La Jolla, California 92093-0653, USA

    • Tingting Du
    •  & Bing Ren
  6. Life Science Center, Tsukuba Advanced Research Alliance, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8577, Japan

    • Yuta Takahashi
  7. Grado en Medicina, Universidad Católica, San Antonio de Murcia, Campus de los Jerónimos, 135, Guadalupe 30107, Spain

    • Jeronimo Lajara
    •  & Pedro Guillen
  8. Fundacion Pedro Guillen, Clínica Cemtro, Avenida Ventisquero de la Condesa, 42, 28035 Madrid, Spain

    • Pedro Guillen
  9. Hospital Clinic of Barcelona, Carrer Villarroel, 170, 08036 Barcelona, Spain

    • Josep M. Campistol
  10. University of California, Davis, Davis, California 95616, USA

    • Pablo J. Ross
  11. The Salk Institute for Biological Studies, Peptide Biology Laboratory, La Jolla, California 92037, USA

    • Alan Saghatelian

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Contributions

J.W., D.O. and J.C.I.B. conceived the study. J.W. and D.O. derived mESC, EpiSC and rsEpiSC lines. J.W., D.O. and C.R.E. designed and performed in vivo embryo grafting experiments. J.W., D.O., M.L., K.S., L.M., Z.L., T.H. and P.R. designed and performed all in vitro experiments; J.M.C., J.L. and P.G. helped project design and discussions and performed microarray experiments. I.T., Y.T. performed bisulfite sequencing experiments; M.K. performed teratoma studies; C.L., Y.H., Z.Z., J.R.N. and J.E. performed whole-genome bisulfite sequencing experiments and analysed data. T.D. and B.R. performed ChIP-seq experiments. C.B. and M.L. performed bioinformatics analysis. A.S. analysed global metabolic profiling data; E.A. and N.K. provided technical support. J.W., D.O., M.L. and J.C.I.B. prepared the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Juan Carlos Izpisua Belmonte.

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    Supplementary Information

    This file contains Supplementary Figure 1 and Western blots for Figure 4b and Extended Data Figure 4c.

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    This file contains Supplementary Tables 1-8.

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https://doi.org/10.1038/nature14413

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