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Human Embryonic Stem Cells Do Not Change Their X Inactivation Status during Differentiation
Sanjeet Patel1, Giancarlo Bonora1, Anna Sahakyan1
1Department of Biological Chemistry, Molecular Biology Institute, Jonsson Comprehensive Cancer Center, Bioinformatics Program, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Cell Reports
|December 20, 2016
Summary
The X chromosome state in human embryonic stem cells (ESCs) remains unchanged during differentiation. This suggests X chromosome dosage compensation is not essential for ESC differentiation, with only XiXIST+Xa ESCs being suitable for applications.
Area of Science:
- Cell Biology
- Genetics
- Epigenetics
Background:
- Embryonic stem cell (ESC) applications depend on accurate chromatin modifications during differentiation.
- The behavior of the X chromosome during ESC differentiation is not well understood.
- Human ESCs exhibit diverse X chromosome states: two active (XaXa), one active and one inactive (XiXIST+Xa;XiXa), or one active and an eroded inactive X (XeXa).
Purpose of the Study:
- To investigate the stability of X chromosome states during human ESC differentiation.
- To determine if X chromosome dosage compensation is necessary for ESC differentiation.
- To identify the most suitable X chromosome state in ESCs for downstream applications.
Main Methods:
- Establishment of human ESC lines with distinct X chromosome states (XiXa, XeXa, XaXa).
- Tracking of X chromosome status throughout the differentiation process.
- Analysis of XIST RNA expression and X-linked gene expression in differentiated cells.
Main Results:
- The pre-existing X chromosome state in primed ESCs is maintained after differentiation.
- Differentiated XeXa and XaXa cells did not exhibit XIST expression or induce X inactivation.
- Differentiated XeXa and XaXa cells showed higher X-linked gene expression compared to XiXa cells.
Conclusions:
- X chromosome dosage compensation is not a requirement for successful ESC differentiation.
- The XiXIST+Xa ESC state is optimal for ESC applications.
- Other X chromosome states (XaXa, XeXa) in ESCs represent aberrant outcomes of derivation and propagation methods.
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