Letter | Published:

Reconstitution in vitro of the entire cycle of the mouse female germ line

Nature volume 539, pages 299303 (10 November 2016) | Download Citation


The female germ line undergoes a unique sequence of differentiation processes that confers totipotency to the egg1,2. The reconstitution of these events in vitro using pluripotent stem cells is a key achievement in reproductive biology and regenerative medicine. Here we report successful reconstitution in vitro of the entire process of oogenesis from mouse pluripotent stem cells. Fully potent mature oocytes were generated in culture from embryonic stem cells and from induced pluripotent stem cells derived from both embryonic fibroblasts and adult tail tip fibroblasts. Moreover, pluripotent stem cell lines were re-derived from the eggs that were generated in vitro, thereby reconstituting the full female germline cycle in a dish. This culture system will provide a platform for elucidating the molecular mechanisms underlying totipotency and the production of oocytes of other mammalian species in culture.

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Gene Expression Omnibus

Data deposits

The RNA-seq data have been deposited at Gene Expression Omnibus (GEO) database under accession number GSE79729.


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We thank Y. Takada for technical support, H. Ohta for technical advice on the iPSCs and embryo transfer, Y. Ohkawa for technical assistance on the RNA-seq, K. Kitajima and C. Meno for providing microscopes, F. Arai for providing FACSAriaII, and H. Leitch for proofreading the manuscript. We also thank the Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences for technical assistance. N.H. was a JSPS Research Fellow. This study was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (KAKENHI #25114006); by JST-PRESTO; by the Uehara Memorial Foundation; and by the Takeda Science Foundation.

Author information

Author notes

    • Orie Hikabe
    •  & Katsuhiko Hayashi

    These authors contributed equally to this work.


  1. Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka 812-8582, Japan

    • Orie Hikabe
    • , Nobuhiko Hamazaki
    • , Go Nagamatsu
    • , Norio Hamada
    • , So Shimamoto
    • , Takuya Imamura
    • , Kinichi Nakashima
    •  & Katsuhiko Hayashi
  2. Department of Bioscience, Tokyo University of Agriculture, 1-1-1, Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan

    • Yayoi Obata
  3. NARO Institute of Livestock and Grassland Science, Ikenodai 2, Tsukuba 305-0901, Japan

    • Yuji Hirao
  4. Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka 812-8582, Japan

    • Norio Hamada
  5. Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan

    • Mitinori Saitou
  6. Center for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan

    • Mitinori Saitou
  7. Institute for Integrated Cell-Material Sciences, Kyoto University, Yoshida-Ushinomiya-cho, Sakyo-ku, Kyoto 606-8501, Japan

    • Mitinori Saitou
  8. JST, ERATO, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan

    • Mitinori Saitou
  9. JST, PRESTO, Maidashi 3-1-1, Higashi-ku, Fukuoka 812-8582, Japan

    • Katsuhiko Hayashi


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O.H., N.H., S.S. and K.H. performed the culture, embryo transfer, immunofluorescence, PCR analysis and chimaera analysis; O.H., N.H., S.S. and K.H. performed the differentiation to mature oocytes and are able to replicate the entire process, independently. N.H., T.I., and K.N. performed the RNA-seq analysis. G.N. generated the iPSC lines. Y.O and Y.H. conducted the assessment of the culture conditions. M.S. contributed to the inception of the study. K.H. designed the experiments and wrote the manuscript.

Corresponding author

Correspondence to Katsuhiko Hayashi.

Reviewer Information Nature thanks D. Egli, S. Mitalipov and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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    Supplementary Information

    This file contains Supplementary Figure 1 (gel source data) and Supplementary Tables 1-2.

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