Inductive and Selective Effects of GSK3 and MEK Inhibition on Nanog Heterogeneity in Embryonic Stem Cells
| Autor: | Fabian J. Theis, Andreas Reimann, Dirk Loeffler, Simon Hastreiter, Daniel L. Coutu, Konstantinos D. Kokkaliaris, Oliver Hilsenbeck, Philipp S. Hoppe, Stavroula Skylaki, Michael Schwarzfischer, Konstantinos Anastassiadis, Timm Schroeder |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
2i MAP Kinase Kinase 2 Gene Expression Gsk3 Biochemistry Nanog Gene Knockout Techniques Glycogen Synthase Kinase 3 Mice continuous time-lapse imaging lcsh:QH301-705.5 reproductive and urinary physiology lcsh:R5-920 cell fate Kinase Mek embryonic stem cells pluripotency cell heterogeneity single-cell tracking Cell Differentiation Nanog Homeobox Protein Cell Fate Cell Heterogeneity Continuous Time-lapse Imaging Embryonic Stem Cells Pluripotency Single-cell Tracking Cell biology embryonic structures Single-Cell Analysis biological phenomena cell phenomena and immunity lcsh:Medicine (General) Signal Transduction Homeobox protein NANOG Programmed cell death Cell fate determination Biology Article Cell Line 03 medical and health sciences Genetics Animals Viability assay Protein Kinase Inhibitors urogenital system Dual effect Cell Biology Embryonic stem cell 030104 developmental biology lcsh:Biology (General) Leukemia inhibitory factor Developmental Biology |
| Zdroj: | Stem Cell Reports, 10 Stem Cell Reports, Vol 11, Iss 1, Pp 58-69 (2018) Stem Cell Reports Stem Cell Rep. 10, 58-69 (2018) |
| DOI: | 10.3929/ethz-b-000274995 |
| Popis: | Summary 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. Graphical Abstract Highlights • Continuous long-term single-cell quantification of 2i effects on murine ESCs • 2i enriches for a Nanog high population through a selective cell death effect • 2i also upregulates Nanog expression and prevents its downregulation • The viability of Nanog−/− cells is compromised in 2i Schroeder and colleagues show that GSK3 and MEK inhibition (2i) has several simultaneous effects on murine ESC cultures. They applied long-term time-lapse imaging to quantify cell death, proliferation, and Nanog expression dynamics in single ESCs. This demonstrated that 2i treatment of ESCs leads to more uniform and high Nanog expression due to both selective and inductive effects. |
| Databáze: | OpenAIRE |
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