Expansion of Mouse Primordial Germ Cell-like Cells Recapitulates an Epigenetic Blank Slate

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2017 Jun 1

PMID 28559416

Citations 55

Authors

Hiroshi Ohta

Kazuki Kurimoto

Ikuhiro Okamoto

Tomonori Nakamura

Yukihiro Yabuta

Hidetaka Miyauchi

Takuya Yamamoto

Yukiko Okuno

Masatoshi Hagiwara

Kenjiro Shirane

Hiroyuki Sasaki

Mitinori Saitou

Affiliations

Soon will be listed here.

Abstract

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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