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Updated: Jun 10, 2026
Efficient Neural Differentiation using Single-Cell Culture of Human Embryonic Stem Cells
Published on: January 18, 2020
Inhibition of ERK1/2 prevents neural and mesendodermal differentiation and promotes human embryonic stem cell
Jie Na1, Miho K Furue, Peter W Andrews
1School of Medicine, Tsinghua University, Beijing, China. obermair@hifo.uzh.ch
Abstract:
Extracellular signal-regulated kinases (ERKs) have many important functions during embryogenesis. However, their role in embryonic stem (ES) cells is controversial. Previous studies reported that, in contrast to mouse ES cells, human ES cells differentiate if ERK1/2 is inhibited. We reexamined the role of ERK1/2 in human ES cells using a chemically defined culture system and found that when ERK1/2 is blocked with specific chemical inhibitors, neural and mesendodermal differentiation is prevented, but cells become sensitive to BMP-induced differentiation. Inhibition of ERK1/2 significantly reduced the clonogenicity of human ES cells by preventing cell adhesion and survival. When this negative effect was avoided, we were able to maintain human ES cell self-renewal for more than 3months in the presence of ERK1/2 inhibitors in a chemically defined culture system containing FGF2 and activin A but no BMP4. Our results suggest that the functional outcome of FGF/ERK1/2 signaling in human ES cells is influenced by the relative levels of activin A/TGFbeta and BMP activity. Moreover, activation of ERK1/2 in human ES cells is required for proper neural and mesendodermal differentiation. In contrast to mouse ES cells, a low level of BMP4 is sufficient to initiate extraembryonic differentiation when ERK1/2 is inhibited.
Insights
In human embryonic stem cells, blocking extracellular signal-regulated kinases (ERKs) prevents differentiation but impairs self-renewal. Maintaining ERK1/2 activity is crucial for neural and mesendodermal development.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Cell Signaling
Background:
- Extracellular signal-regulated kinases (ERKs) play critical roles in embryogenesis, but their function in human embryonic stem cells (hESCs) remains debated.
- Previous research indicated that inhibiting ERK1/2 in hESCs leads to differentiation, contrasting with findings in mouse ES cells.
Purpose of the Study:
- To re-evaluate the role of ERK1/2 signaling in hESCs within a chemically defined culture system.
- To investigate the impact of ERK1/2 inhibition on hESC differentiation, self-renewal, and response to growth factors.
Main Methods:
- Utilized a chemically defined culture system for hESCs.
- Employed specific chemical inhibitors to block ERK1/2 activity.
- Assessed differentiation, clonogenicity, cell adhesion, and survival under various culture conditions.
Main Results:
- ERK1/2 inhibition prevented neural and mesendodermal differentiation but increased sensitivity to BMP-induced differentiation.
- Inhibition of ERK1/2 reduced hESC clonogenicity by impairing cell adhesion and survival.
- Sustained hESC self-renewal (>3 months) was achieved with ERK1/2 inhibitors by managing FGF2, activin A, and BMP4 levels.
Conclusions:
- The effect of FGF/ERK1/2 signaling in hESCs is modulated by the balance of activin A/TGFbeta and BMP signaling.
- ERK1/2 activation is essential for appropriate neural and mesendodermal differentiation in hESCs.
- Unlike mouse ES cells, low BMP4 levels can induce extraembryonic differentiation when ERK1/2 is inhibited in hESCs.
