Journal
DEVELOPMENT
Volume 143, Issue 16, Pages 2958-2964Publisher
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/dev.136739
Keywords
X chromosome reactivation; X chromosome inactivation; Live-cell imaging; Pluripotent stem cells; Early mouse development
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Funding
- Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology (PRESTO)
- Ministry of Education, Culture, Sports, Science, and Technology [23500492, 15H01468]
- Grants-in-Aid for Scientific Research [15H05573, 23500492, 15K18456, 25712035, 15H01468] Funding Source: KAKEN
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Pluripotent stem cells can be classified into two distinct states, naive 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 naive 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.
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