title: Pluripotent stem cells derived from mouse primordial germ cells by small molecule compounds.
abstract: Primordial germ cells (PGCs) can give rise to pluripotent stem cells known as embryonic germ cells (EGCs) when cultured with basic fibroblast growth factor (bFGF), stem cell factor (SCF), and leukemia inhibitory factor. Somatic cells can give rise to induced pluripotent stem cells (iPSCs) by introduction of the reprogramming transcription factors Oct4, Sox2, and Klf4. The effects of Sox2 and Klf4 on somatic cell reprogramming can be reproduced using the small molecule compounds, transforming growth factor-β receptor (TGFβR) inhibitor and Kempaullone, respectively. Here we examined the effects of TGFβR inhibitor and Kempaullone on EGC derivation from PGCs. Treatment of PGCs with TGFβR inhibitor and/or Kempaullone generated pluripotent stem cells under standard embryonic stem cell (ESC) culture conditions without bFGF and SCF, which we termed induced EGCs (iEGCs). The derivation efficiency of iEGCs was dependent on the differentiation stage and sex. DNA methylation levels of imprinted genes in iEGCs were reduced, with the exception of the H19 gene. The promoters of genes involved in germline development were generally hypomethylated in PGCs, but three germline genes showed comparable DNA methylation levels among iEGs, ESCs, and iPSCs. These results show that PGCs can be reprogrammed into pluripotent state using small molecule compounds, and that DNA methylation of these germline genes is not maintained in iEGCs.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan; Department of Pathology, Osaka University, Suita, Osaka, Japan; Laboratory of Molecular Embryology, Kitasato University School of Science, Kitasato, Minami-ku, Sagamihara, Kanagawa, Japan; Laboratory of Stem Cell Biology, 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: Yoichi
authors.initials: Y
authors.lastname: Sekita
authors.forename: Keita
authors.initials: K
authors.lastname: Fujikawa
authors.forename: Tsunetoshi
authors.initials: T
authors.lastname: Nakatani
authors.forename: Mika
authors.initials: M
authors.lastname: Odamoto
authors.forename: Soichiro
authors.initials: S
authors.lastname: Funaki
authors.forename: Masahito
authors.initials: M
authors.lastname: Ikawa
authors.forename: Kuniya
authors.initials: K
authors.lastname: Abe
authors.forename: Toru
authors.initials: T
authors.lastname: Nakano
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D019175:DNA Methylation; D004149:Dioxoles / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D057890:Epigenomics; D005260:Female; D005854:Germ Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D059467:Transcriptome
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4
keywords: DNA methylation; Embryonic germ cells; Epigenetic memory; Induced pluripotent stem cells; Primordial germ cells; Small molecule compound