TY - JOUR
T1 - In vitro expansion of mouse primordial germ cell-like cells recapitulates an epigenetic blank slate
AU - Ohta, Hiroshi
AU - Kurimoto, Kazuki
AU - Okamoto, Ikuhiro
AU - Nakamura, Tomonori
AU - Yabuta, Yukihiro
AU - Miyauchi, Hidetaka
AU - Yamamoto, Takuya
AU - Okuno, Yukiko
AU - Hagiwara, Masatoshi
AU - Shirane, Kenjiro
AU - Sasaki, Hiroyuki
AU - Saitou, Mitinori
N1 - Funding Information:
We thank the members of our laboratory for their helpful input on this study, and Y. Nagai, R. Kabata, N. Konishi, Y. Sakaguchi, and M. Kawasaki of the Saitou Laboratory, M. Miyake, T. Akinaga, and J. Oishi of the Sasaki Laboratory, and T. Sato and M. Kabata of the Yamamoto Laboratory for their technical assistance. Chemical compounds for the screening were provided in part from the Medical Research Support Center, Graduate School of Medicine, Kyoto University, which was supported by Platform for Drug Discovery, Informatics, and Structural Life Science from the Ministry of Education, Culture, Sports, Science and Technology, Japan. This work was supported in part by a Grant-in-Aid from JSPS to H.O. (JP24680045 and JP15H05636), and to K.K. (JP16H04720 and JP16H01216) by a Grant-in-Aid for Scientific Research on Innovative Areas from JSPS to H.S. (JP25112010), and by JST-ERATO to M.S. (JPMJER1104).
Publisher Copyright:
© 2017 The Authors
PY - 2017/7/3
Y1 - 2017/7/3
N2 - The expansion of primordial germ cells (PGCs), the precursors for the oocytes and spermatozoa, is a key challenge in reproductive biology/medicine. Using a chemical screening exploiting PGC-like cells (PGCLCs) induced from mouse embryonic stem cells (ESCs), we here identify key signaling pathways critical for PGCLC proliferation. We show that the combinatorial application of Forskolin and Rolipram, which stimulate cAMP signaling via different mechanisms, expands PGCLCs up to ~50-fold in culture. The expanded PGCLCs maintain robust capacity for spermatogenesis, rescuing the fertility of infertile mice. Strikingly, during expansion, PGCLCs comprehensively erase their DNA methylome, including parental imprints, in a manner that precisely recapitulates genome-wide DNA demethylation in gonadal germ cells, while essentially maintaining their identity as sexually uncommitted PGCs, apparently through appropriate histone modifications. By establishing a paradigm for PGCLC expansion, our system reconstitutes the epigenetic “blank slate” of the germ line, an immediate precursory state for sexually dimorphic differentiation.
AB - The expansion of primordial germ cells (PGCs), the precursors for the oocytes and spermatozoa, is a key challenge in reproductive biology/medicine. Using a chemical screening exploiting PGC-like cells (PGCLCs) induced from mouse embryonic stem cells (ESCs), we here identify key signaling pathways critical for PGCLC proliferation. We show that the combinatorial application of Forskolin and Rolipram, which stimulate cAMP signaling via different mechanisms, expands PGCLCs up to ~50-fold in culture. The expanded PGCLCs maintain robust capacity for spermatogenesis, rescuing the fertility of infertile mice. Strikingly, during expansion, PGCLCs comprehensively erase their DNA methylome, including parental imprints, in a manner that precisely recapitulates genome-wide DNA demethylation in gonadal germ cells, while essentially maintaining their identity as sexually uncommitted PGCs, apparently through appropriate histone modifications. By establishing a paradigm for PGCLC expansion, our system reconstitutes the epigenetic “blank slate” of the germ line, an immediate precursory state for sexually dimorphic differentiation.
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U2 - 10.15252/embj.201695862
DO - 10.15252/embj.201695862
M3 - Article
C2 - 28559416
AN - SCOPUS:85019681041
SN - 0261-4189
VL - 36
SP - 1888
EP - 1907
JO - EMBO Journal
JF - EMBO Journal
IS - 13
ER -