title: Tracing the conversion process from primordial germ cells to pluripotent stem cells in mice.
abstract: To understand mechanisms underlying acquisition of pluripotency, it is critical to identify cells that can be converted to pluripotent stem cells. For this purpose, we focused on unipotent primordial germ cells (PGCs), which can be reprogrammed into pluripotent embryonic germ (EG) cells under defined conditions. Treatment of PGCs with combinations of signaling inhibitors, including inhibitors of MAP2K (MEK), GSK3B (GSK-3beta), and TGFB (TGFbeta) type 1 receptors, induced cells to enter a pluripotent state at a high frequency (12.1%) by Day 10 of culture. When we employed fluorescence-activated cell sorting to monitor conversion of candidate cells to a pluripotent state, we observed a cell cycle shift to S phase, indicating enrichment of pluripotent cells, during the early phase of EG formation. Transcriptome analysis revealed that PGCs retained expression of some pluripotent stem cell-associated genes, such as Pou5f1 and Sox2, during EG cell formation. On the other hand, PGCs lost their germ lineage characteristics and acquired expression of pluripotent stem cell markers, such as Klf4 and Eras. The overall gene expression profiles revealed by this system provide novel insight into how pluripotency is acquired in germ-committed cells.
authors.affiliation: Department of Cell Differentiation, The Sakaguchi Laboratory, School of Medicine, Keio University, Tokyo, Japan. gonag@z2.keio.jp
authors.forename: Go
authors.initials: G
authors.lastname: Nagamatsu
authors.forename: Takeo
authors.initials: T
authors.lastname: Kosaka
authors.forename: Shigeru
authors.initials: S
authors.lastname: Saito
authors.forename: Keiyo
authors.initials: K
authors.lastname: Takubo
authors.forename: Hideo
authors.initials: H
authors.lastname: Akiyama
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Sudo
authors.forename: Katsuhisa
authors.initials: K
authors.lastname: Horimoto
authors.forename: Mototsugu
authors.initials: M
authors.lastname: Oya
authors.forename: Toshio
authors.initials: T
authors.lastname: Suda
mesh_terms: D000818:Animals; D001549:Benzamides; D018929:Cell Culture Techniques; D004159:Diphenylamine / Q000031:analogs & derivatives; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005854:Germ Cells; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors; D000090062:Kruppel-Like Factor 4; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D008819:Mice, Nude; D008822:Mice, Transgenic; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells; D011725:Pyridines; D011743:Pyrimidines; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors; D016196:S Phase
chemical_list: D001549:Benzamides; C473711:Chir 99021; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D011725:Pyridines; D011743:Pyrimidines; D018125:Receptors, Transforming Growth Factor beta; C506614:mirdametinib; D004159:Diphenylamine; D038362:Glycogen Synthase Kinase 3; D020929:Mitogen-Activated Protein Kinase Kinases