Human primordial germ cell commitment in vitro associates with a unique PRDM14 expression profile

• Published in: The EMBO journal Vol. 34; no. 8; pp. 1009 - 1024
• Main Authors: Sugawa, Fumihiro, Araúzo-Bravo, Marcos J, Yoon, Juyong, Kim, Kee-Pyo, Aramaki, Shinya, Wu, Guangming, Stehling, Martin, Psathaki, Olympia E, Hübner, Karin, Schöler, Hans R
• Format: Journal Article
• Language: English
• Published: London Blackwell Publishing Ltd 15-04-2015; Nature Publishing Group UK; BlackWell Publishing Ltd
• Subjects: Activins - pharmacology; Animals; BLIMP1; Bone Morphogenetic Protein 4 - pharmacology; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cells, Cultured; Cellular biology; DNA methylation; EMBO11; EMBO22; EMBO39; Epigenesis, Genetic; Fibroblast Growth Factor 2 - pharmacology; Gene expression; Germ Cells - cytology; Germ Cells - physiology; human pluripotent stem cells; Humans; Mice; Microarray Analysis; Pluripotent Stem Cells - drug effects; Pluripotent Stem Cells - physiology; primordial germ cell precursors; primordial germ cell specification; Repressor Proteins - genetics; Stem cells; Transcriptome - drug effects; human pluripotent stem cells; primordial germ cell specification; BLIMP1; primordial germ cell precursors
• ISSN: 0261-4189; 1460-2075
• DOI: 10.15252/embj.201488049

Primordial germ cells (PGCs) develop only into sperm and oocytes
in vivo
. The molecular mechanisms underlying human PGC specification are poorly understood due to inaccessibility of cell materials and lack of
in vitro
models for tracking the earliest stages of germ cell development. Here, we describe a defined and stepwise differentiation system for inducing pre‐migratory PGC‐like cells (PGCLCs) from human pluripotent stem cells (PSCs). In response to cytokines, PSCs differentiate first into a heterogeneous mesoderm‐like cell population and then into PGCLCs, which exhibit minimal PRDM14 expression. PGC specification in humans is similar to the murine process, with the sequential activation of mesodermal and PGC genes, and the suppression of neural induction and of
de novo
DNA methylation, suggesting that human PGC formation is induced via epigenesis, the process of germ cell specification via inductive signals from surrounding somatic cells. This study demonstrates that PGC commitment in humans shares key features with that of the mouse, but also highlights key differences, including transcriptional regulation during the early stage of human PGC development (3–6 weeks). A more comprehensive understanding of human germ cell development may lead to methodology for successfully generating PSC‐derived gametes for reproductive medicine.
Synopsis
Primordial germ cells (PGCs) generate sperm and oocytes. A defined differentiation system induces human pre‐migratory PGC‐like cells (PGCLCs)
in vitro
from pluripotent stem cells (PSCs). This protocol allows disease modeling and may lead to the generation of gametes for reproductive medicine.
PSCs differentiate first into a heterogeneous mesoderm‐like cell population and then into PGCLCs.
PGCLCs exhibit minimal expression of the PGC gene
PRDM14
.
PGC specification in humans is similar to the murine process in that mesodermal and PGC genes are sequentially activated and neural induction and
de novo
DNA methylation are suppressed.
Differences to the murine process include transcriptional regulation during the early stages of human PGC development (3–6 weeks).
Graphical Abstract
Primordial germ cells (PGCs) generate sperm and oocytes. A defined differentiation system induces human pre‐migratory PGC‐like cells (PGCLCs) from pluripotent stem cells (PSCs)
in vitro
. This protocol allows disease modeling and may lead to the generation of gametes for reproductive medicine.