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Updated: Jul 4, 2026
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Combined DNA-RNA Fluorescent In situ Hybridization (FISH) to Study X Chromosome Inactivation in Differentiated Female Mouse Embryonic Stem Cells
Published on: June 14, 2014
X chromosome inactivation in nuclear transfer ES cells
S Shibata1, T Wakayama, T Yokota
1Department of Stem Cell Biology, Graduate School of Medicine, Kanazawa University, Kanazawa, Japan. shinwa@med.kanazawa-u.ac.jp
Cytogenetic and Genome Research
|June 12, 2008
Summary
Nuclear transfer ES (ntES) cells show normal X chromosome inactivation (XCI), unlike cloned mice. This suggests epigenetic reprogramming for safe therapeutic cloning applications.
Area of Science:
- Stem cell biology
- Epigenetics
- Mammalian reproductive technology
Background:
- Nuclear transfer ES (ntES) cells are derived from cloned blastocysts and hold promise for regenerative medicine.
- Cloned mammals often exhibit abnormalities, indicating potential issues with gene regulation and epigenetic reprogramming.
- Previous studies noted random X chromosome inactivation (XCI) in cloned embryos, not always aligning with Xce strength, suggesting incomplete epigenetic reprogramming.
Purpose of the Study:
- To investigate X chromosome inactivation (XCI) in nuclear transfer ES (ntES) cell lines.
- To determine if epigenetic marks related to XCI are properly reprogrammed during ntES cell establishment.
- To assess the suitability of ntES cells for therapeutic cloning by examining XCI patterns.
Main Methods:
- Established ntES cell lines from differentiated embryoid bodies of a female mouse ES cell line.
- Examined Xist RNA localization, histone modifications at the Xist locus, and XCI choice in ntES cells.
- Compared XCI patterns in ntES cells with their parental ES cell line.
Main Results:
- No substantial differences were observed between ntES cell lines and the parental ES cell line regarding Xist RNA localization and histone modifications.
- ntES cells exhibited normal XCI choice, consistent with the parental ES line.
- These findings suggest successful reprogramming of the Xist locus and associated epigenetic marks during nuclear transfer and ntES cell derivation.
Conclusions:
- Nuclear transfer and subsequent ntES cell establishment effectively reprogram epigenetic marks involved in XCI.
- Normal XCI choice in ntES cells contrasts with skewed XCI in cloned mice, supporting their potential for safe therapeutic cloning.
- The findings bolster the goal of utilizing ntES cells for clinical applications in regenerative medicine.
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