Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells.pdf

Reconstitution of the Mouse Germ Cell Specification Pathway in Culture by Pluripotent Stem Cells Katsuhiko Hayashi,1,3 Hiroshi Ohta,1,3 Kazuki Kurimoto,1,3 Shinya Aramaki,1 and Mitinori Saitou1,2,3,* 1Department of Anatomy and Cell Biology, Graduate School of Medicine 2Institute for Integrated Cell-Material Sciences Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan 3JST, CREST, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan *Correspondence: saitou@anat2.med.kyoto-u.ac.jp DOI 10.1016/j.cell.2011.06.052SUMMARY the PGC fate (Kurimoto et al., 2008; Ohinata et al., 2005; VincentThe generation of properly functioning gametes in vitro requires reconstitution of the multistepped pathway of germ cell development. We demonstrate here the generation of primordial germ cell-like cells (PGCLCs) in mice with robust capacity for spermato- genesis. PGCLCs were generated from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) through epiblast-like cells (EpiLCs), a cellular state highly similar to pregastrulating epiblasts but distinct from epiblast stem cells (EpiSCs). Reflecting epiblast development, EpiLC induction from ESCs/ iPSCs is a progressive process, and EpiLCs highly competent for the PGC fate are a transient entity. The global transcription profiles, epigenetic reprog- ramming, and cellular dynamics during PGCLC induction from EpiLCs meticulously capture those associated with PGC specification from the epi- blasts. Furthermore, we identify Integrin-b3 and SSEA1 asmarkers that allow the isolation of PGCLCs with spermatogenic capacity from tumorigenic un- differentiated cells. Our findings provide a paradigm for the first step of in vitro gametogenesis. INTRODUCTION The germ cell lineage ensures the creation of new individuals in most multicellular organisms, perpetuating the genetic and epigenetic information across the generations. Accordingly, in vitro reconstitution of germ cell development is one of the most fundamental chall