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In vitro expansion of mouse primordial germ cell‐like cells recapitulates an epigenetic blank slate
Author(s) -
Ohta Hiroshi,
Kurimoto Kazuki,
Okamoto Ikuhiro,
Nakamura Tomonori,
Yabuta Yukihiro,
Miyauchi Hidetaka,
Yamamoto Takuya,
Okuno Yukiko,
Hagiwara Masatoshi,
Shirane Kenjiro,
Sasaki Hiroyuki,
Saitou Mitinori
Publication year - 2017
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201695862
Subject(s) - biology , epigenetics , dna methylation , microbiology and biotechnology , germ cell , chromatin , genetics , dna , gene expression , gene
The expansion of primordial germ cells ( PGC s), the precursors for the oocytes and spermatozoa, is a key challenge in reproductive biology/medicine. Using a chemical screening exploiting PGC ‐like cells ( PGCLC s) induced from mouse embryonic stem cells ( ESC s), 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 PGCLC s up to ~50‐fold in culture. The expanded PGCLC s maintain robust capacity for spermatogenesis, rescuing the fertility of infertile mice. Strikingly, during expansion, PGCLC s 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 PGC s, 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.