title: Induction of primordial germ cell-like cells from mouse embryonic stem cells by ERK signal inhibition.
abstract: Primordial germ cells (PGCs) are embryonic germ cell precursors. Specification of PGCs occurs under the influence of mesodermal induction signaling during in vivo gastrulation. Although bone morphogenetic proteins and Wnt signaling play pivotal roles in both mesodermal and PGC specification, the signal regulating PGC specification remains unknown. Coculture of mouse embryonic stem cells (ESCs) with OP9 feeder cells induces mesodermal differentiation in vitro. Using this mesodermal differentiation system, we demonstrated that PGC-like cells were efficiently induced from mouse ESCs by extracellular signal-regulated kinase (ERK) signaling inhibition. Inhibition of ERK signaling by a MAPK/ERK kinase (MEK) inhibitor upregulated germ cell marker genes but downregulated mesodermal genes. In addition, the PGC-like cells showed downregulation of DNA methylation and formed pluripotent stem cell colonies upon treatment with retinoic acid. These results show that inhibition of ERK signaling suppresses mesodermal differentiation but activates germline differentiation program in this mesodermal differentiation system. Our findings provide a new insight into the signaling networks regulating PGC specification.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan; Department of Pathology, Medical School, Osaka University, Suita, Osaka, Japan; Laboratory of Molecular Embryology, Department of Biosciences, Kitasato University School of Science, Kitasato, Minami-ku, Sagamihara, Kanagawa, Japan; Laboratory of Stem Cell Biology, Department of Biosciences, Kitasato University School of Science, Kitasato, Minami-ku, Sagamihara, Kanagawa, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Kimura
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kaga
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ohta
authors.forename: Mika
authors.initials: M
authors.lastname: Odamoto
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Sekita
authors.forename: Kunpeng
authors.initials: K
authors.lastname: Li
authors.forename: Noriko
authors.initials: N
authors.lastname: Yamano
authors.forename: Keita
authors.initials: K
authors.lastname: Fujikawa
authors.forename: Ayako
authors.initials: A
authors.lastname: Isotani
authors.forename: Norihiko
authors.initials: N
authors.lastname: Sasaki
authors.forename: Masashi
authors.initials: M
authors.lastname: Toyoda
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Hayashi
authors.forename: Masaru
authors.initials: M
authors.lastname: Okabe
authors.forename: Takashi
authors.initials: T
authors.lastname: Shinohara
authors.forename: Mitinori
authors.initials: M
authors.lastname: Saitou
authors.forename: Toru
authors.initials: T
authors.lastname: Nakano
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology*; D044127:Epigenesis, Genetic / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D061252:Feeder Cells / Q000166:cytology / Q000187:drug effects; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005854:Germ Cells / Q000166:cytology* / Q000201:enzymology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D013091:Spermatogenesis / Q000187:drug effects; D033581:Stem Cell Transplantation; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D047428:Protein Kinase Inhibitors; D014212:Tretinoin; D048049:Extracellular Signal-Regulated MAP Kinases; D020929:Mitogen-Activated Protein Kinase Kinases
keywords: ERK signal; Embryonic stem cells; In vitro differentiation; Mesoderm; Primordial germ cells