Reconstitution of mouse oogenesis in a dish from pluripotent stem cells

Katsuhiko Hayashi, Orie Hikabe, Yayoi Obata, Yuji Hirao

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53 Citations (Scopus)


Generation of functional oocytes in culture from pluripotent stem cells should provide a useful model system for improving our understanding of the basic mechanisms underlying oogenesis. In addition, it has potential applications as an alternative source of oocytes for reproduction. Using the most advanced mouse model in regard to reproductive engineering and stem cell biology, we previously developed a culture method that produces functional primorial germ cells starting from pluripotent cells in culture and described it in a previous protocol. This Protocol Extension describes an adaptation of this existing Protocol in which oogenesis also occurs in vitro, thus substantially modifying the technique. Oocytes generated from embryonic stem cells (ESCs) or induced pluripotent stem cells give rise to healthy pups. Here, we describe the protocol for oocyte generation in culture. The protocol is mainly composed of three different culture stages: in vitro differentiation (IVDi), in vitro growth (IVG), and in vitro maturation (IVM), which in total take ∼5 weeks. In each culture period, there are several checkpoints that enable the number of oocytes being produced in the culture to be monitored. The basic structure of the culture system should provide a useful tool for clarifying the complicated sequence of oogenesis in mammals.

Original languageEnglish
Pages (from-to)1733-1744
Number of pages12
JournalNature Protocols
Issue number9
Publication statusPublished - Sept 1 2017

All Science Journal Classification (ASJC) codes

  • General Biochemistry,Genetics and Molecular Biology


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