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Inductive and selective effects of GSK3 and MEK inhibition on nanog heterogeneity in embryonic stem cells.
Stem Cell Rep. 10, 58-69 (2018)
Embryonic stem cells (ESCs) display heterogeneous expression of pluripotency factors such as Nanog when cultured with serum and leukemia inhibitory factor (LIF). In contrast, dual inhibition of the signaling kinases GSK3 and MEK (2i) converts ESC cultures into a state with more uniform and high Nanog expression. However, it is so far unclear whether 2i acts through an inductive or selective mechanism. Here, we use continuous time-lapse imaging to quantify the dynamics of death, proliferation, and Nanog expression in mouse ESCs after 2i addition. We show that 2i has a dual effect: it both leads to increased cell death of Nanog low ESCs (selective effect) and induces and maintains high Nanog levels (inductive effect) in single ESCs. Genetic manipulation further showed that presence of NANOG protein is important for cell viability in 2i medium. This demonstrates complex Nanog-dependent effects of 2i treatment on ESC cultures.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
2i ; Gsk3 ; Mek ; Nanog ; Cell Fate ; Cell Heterogeneity ; Continuous Time-lapse Imaging ; Embryonic Stem Cells ; Pluripotency ; Single-cell Tracking; Ground-state Pluripotency; Self-renewal; Es Cells; Dna Methylation; Expression; Dynamics; Protein; Quantification; Identification; Maintenance
ISSN (print) / ISBN
2213-6711
Journal
Stem Cell Reports
Quellenangaben
Volume: 10,
Issue: 1,
Pages: 58-69
Publisher
Cell Press
Publishing Place
Maryland Heights, MO
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Peer reviewed