title: Live imaging of X chromosome reactivation dynamics in early mouse development can discriminate naïve from primed pluripotent stem cells.
abstract: Pluripotent stem cells can be classified into two distinct states, naïve and primed, which show different degrees of potency. One difficulty in stem cell research is the inability to distinguish these states in live cells. Studies on female mice have shown that reactivation of inactive X chromosomes occurs in the naïve state, while one of the X chromosomes is inactivated in the primed state. Therefore, we aimed to distinguish the two states by monitoring X chromosome reactivation. Thus far, X chromosome reactivation has been analysed using fixed cells; here, we inserted different fluorescent reporter gene cassettes (mCherry and eGFP) into each X chromosome. Using these knock-in 'Momiji' mice, we detected X chromosome reactivation accurately in live embryos, and confirmed that the pluripotent states of embryos were stable ex vivo, as represented by embryonic and epiblast stem cells in terms of X chromosome reactivation. Thus, Momiji mice provide a simple and accurate method for identifying stem cell status based on X chromosome reactivation.
authors.affiliation: Department of Epigenetics, Medical Research Institute, Tokyo Medical & Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan kobayashi.mtt@mri.tmd.ac.jp.
authors.forename: Shin
authors.identifier: 0000-0001-6639-3810
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Department of Epigenetics, Medical Research Institute, Tokyo Medical & Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Hosoi
authors.affiliation: Department of Epigenetics, Medical Research Institute, Tokyo Medical & Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Hirosuke
authors.initials: H
authors.lastname: Shiura
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yamagata
authors.affiliation: Department of Epigenetics, Medical Research Institute, Tokyo Medical & Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Saori
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Fujihara
authors.affiliation: Department of Epigenetics, Medical Research Institute, Tokyo Medical & Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kohda
authors.affiliation: Research Institute for Microbial Diseases, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Okabe
authors.affiliation: Department of Epigenetics, Medical Research Institute, Tokyo Medical & Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Fumitoshi
authors.initials: F
authors.lastname: Ishino
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D005855:Germ Layers / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008817:Mice, Mutant Strains; D010735:Phosphoglycerate Kinase / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D014960:X Chromosome / Q000235:genetics / Q000378:metabolism*; D049951:X Chromosome Inactivation / Q000235:genetics / Q000502:physiology*
chemical_list: C528165:Pgk1 protein, mouse; D010735:Phosphoglycerate Kinase
keywords: Early mouse development; Live-cell imaging; Pluripotent stem cells; X chromosome inactivation; X chromosome reactivation