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Inhibition of mTOR induces a paused pluripotent state
Open accessAbstract
Cultured pluripotent stem cells are a cornerstone of regenerative medicine owing to their ability to give rise to all cell types of the body. Although pluripotent stem cells can be propagated indefinitely in vitro, pluripotency is paradoxically a transient state in vivo, lasting 2-3 days around the time of blastocyst implantation. The exception to this rule is embryonic diapause, a reversible state of suspended development triggered by unfavourable conditions. Diapause is a physiological reproductive strategy widely employed across the animal kingdom, including in mammals, but its regulation remains poorly understood. Here we report that the partial inhibition of mechanistic target of rapamycin (mTOR), a major nutrient sensor and promoter of growth, induces reversible pausing of mouse blastocyst development and allows their prolonged culture ex vivo. Paused blastocysts remain pluripotent and competent-able to give rise to embryonic stem (ES) cells and live, fertile mice. We show that both naturally diapaused blastocysts in vivo and paused blastocysts ex vivo display pronounced reductions in mTOR activity, translation, histone modifications associated with gene activity and transcription. Pausing can be induced directly in cultured ES cells and sustained for weeks without appreciable cell death or deviations from cell cycle distributions. We show that paused ES cells display a remarkable global suppression of transcription, maintain a gene expression signature of diapaused blastocysts and remain pluripotent. These results uncover a new pluripotent stem cell state corresponding to the epiblast of the diapaused blastocyst and indicate that mTOR regulates developmental timing at the peri-implantation stage. Our findings have implications in the fields of assisted reproduction, regenerative medicine, cancer, metabolic disorders and ageing.
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“Similar intermediate states have been described during embryonic development, where cells enter a paused, pluripotent state in response to environmental stress.62”
From Paneth-like transition drives resistance to dual targeting of KRAS and EGFR in colorectal cancer (2026) · cites this paper as [62]
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“For example, of the 131 cells that undergo apoptosis in C. elegans, the great majority (105) are neurons, thus highlighting the importance of PCD for CNS development.22”
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“Lysosomes are also important in apoptosis, a form of programmed cell death (PCD) that is requisite for the appropriate development of the mammalian nervous system.21,22.”
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Metadata
- Authors
- Bulut-Karslioglu, A., Biechele, S., Jin, H., Macrae, T. A., Hejna, M., Gertsenstein, M., Song, J. S., & Ramalho-Santos, M.
- Affiliations
- University of California, San Francisco (US)
- University of Illinois Urbana-Champaign (US)
- Toronto Centre for Phenogenomics (CA)
- Venue
- Nature
- Published
- 2016-11-23
- Volume
- 540
- Issue
- 7631
- Pages
- 119-123
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC5143278
- OpenAlex
- W2553053943
- Cited by
- 283
- References
- 39
- Citations by year
- 2026: 17 · 2025: 40 · 2024: 37 · 2023: 32 · 2022: 32 · 2021: 42 · 2020: 23 · 2019: 27 · 2018: 17 · 2017: 14 · 2016: 2
- FWCI
- 10.40
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- National Science Foundation
- National Institutes of Health (R01OD012204)
- University of California, San Francisco
- National Cancer Institute (P30CA082103, R01CA163336)
- National Institute of General Medical Sciences (R01GM113014)
- National Institute of Diabetes and Digestive and Kidney Diseases (P30DK063720)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Reproductive Medicine › Reproductive Biology and Fertility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- mTOR inhibition, delayed implantation, blastocyst development, pluripotency, epiblast, embryonic stem cells, long-term culture, transcriptional repression, histone modifications, peri-implantation development, developmental timing, assisted reproduction, regenerative medicine, rapamycin, mouse embryo, translational regulation, nutrient sensing
- MeSH terms
- Mouse Embryonic Stem Cells, Animals, Blastocyst, Cells, Cultured, Female, Germ Layers, Histones, Male, Transcription, Genetic, Protein Biosynthesis, Down-Regulation, Pluripotent Stem Cells, Embryonic Development, Mice, TOR Serine-Threonine Kinases, In Vitro Techniques
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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