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Updated: Jun 8, 2026
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An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
Published on: June 1, 2018
Dynamic instability of genomic methylation patterns in pluripotent stem cells
Steen Kt Ooi1, Daniel Wolf, Odelya Hartung
1Department of Genetics and Development, Columbia University, New York, USA. thb12@columbia.edu.
Epigenetics & Chromatin
|September 28, 2010
Summary
DNA methylation is less stable in pluripotent stem cells than in somatic cells, requiring DNMT3L for de novo methylation. This epigenetic instability, especially in female cells, has implications for stem cell therapies.
Area of Science:
- Epigenetics
- Stem Cell Biology
- Genomics
Background:
- Genomic methylation patterns are established during gametogenesis and maintained in somatic cells.
- Previous studies suggested less stable methylation patterns in embryonic stem (ES) cells compared to differentiated somatic cells.
- The mechanisms of de novo and maintenance methylation in pluripotent stem cells versus differentiating somatic cells were unclear.
Purpose of the Study:
- To investigate the mechanisms of de novo and maintenance DNA methylation in pluripotent stem cells.
- To compare DNA methylation dynamics in pluripotent stem cells with those in differentiated somatic cells.
- To assess the role of DNMT3L in DNA methylation in ES cells.
Main Methods:
- Ablation of DNA methyltransferase regulator DNMT3L in mouse ES cells.
- Analysis of de novo methylation of newly integrated retroviral DNA.
- Longitudinal culture of ES cells to assess DNA methylation stability.
- Comparison of methylation loss rates between male and female ES cells.
- Analysis of human ES and induced pluripotent stem cell lines.
Main Results:
- Ablation of DNMT3L in mouse ES cells abolished de novo methylation.
- ES cells lacking DNMT3L exhibited time-dependent DNA methylation loss.
- Wild-type female ES cells showed faster DNA methylation loss than male ES cells.
- Human ES and induced pluripotent stem cell lines displayed significant, variable methylation loss.
- DNMT3L is required for DNA methylation in stem cells but not expressed in differentiating somatic cells.
Conclusions:
- DNA methylation in pluripotent stem cells is dynamic and error-prone, unlike in differentiated cells.
- DNMT3L is essential for DNA methylation in stem cells.
- Epigenetic variability in pluripotent stem cells, particularly in cultured female cells, has negative implications for clinical applications.
- Error-prone maintenance methylation can lead to unpredictable phenotypic variation in clonal stem cell populations.
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