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Cellular and Molecular Life Sciences

, Volume 69, Issue 23, pp 4067–4077 | Cite as

Reestablishment of the inactive X chromosome to the ground state through cell fusion-induced reprogramming

  • Hyun Woo Choi
  • Jong Soo Kim
  • Hyo Jin Jang
  • Sol Choi
  • Jae-Hwan Kim
  • Hans R. Schöler
  • Jeong Tae Do
Research article

Abstract

The restricted gene expression pattern of a differentiated cell can be reversed by fusion of the somatic cell with a more developmentally potent cell type, such as an embryonic stem (ES) cell. During this reprogramming process, somatic cells obtain most of the characteristics of pluripotent cells. Reactivation of an inactive X chromosome (Xi) is an important epigenetic marker confirming the pluripotent reprogramming of somatic cells. Female somatic cells contain one active X chromosome (Xa) and one Xi, and following the fusion of these cells with male ES cells, the Xi becomes activated, resulting in XaXaXaY fusion hybrid cells. To monitor Xi reactivation, transgenic female neural stem cells (fNSCs) carrying a green fluorescent protein (GFP) reporter gene expressed on the Xa (X-GFP), but not on the Xi, were used for reprogramming. XaXiGFP NSCs, whose GFP reporter was silenced, were fused with HM1 ES cells (XY) to induce pluripotent reprogramming. The XiGFP of NSCs were found to be activated on day 4 post-fusion, indicating reactivation of the Xi. Hybrid cells showed pluripotent cell-specific characteristics cells including inactivation of the NSC marker Nestin, DNA demethylation of Oct4, DNA methylation of Nestin, and reactivation of the Xi. Following differentiation of the (GFP-positive) hybrid cells through embryoid body formation, the proportion of GFP-negative cells was found to be approximately 26 %, indicating that there was random inactivation of one of the three Xas. Here, we showed that the Xi of somatic cells is reprogrammed to the Xa state and that cellular differentiation occurs randomly, i.e., regardless of the Xa or Xi state, indicating that the memory of the Xi of somatic cells has been erased and reset to the ground state (i.e., inner cell mass-like state), indicating that random X-chromosome inactivation occurs upon differentiation.

Keywords

Reprogramming Stem cells Cell fusion X-chromosome inactivation Pluripotency 

Notes

Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (Grant 20110019489) and by the Next Generation Bio-Green 21 Program funded by the Rural Development Administration (Grant PJ008009).

Supplementary material

18_2012_1139_MOESM1_ESM.docx (1.1 mb)
Supplementary material 1 (DOCX 1090 kb)

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Copyright information

© Springer Basel AG 2012

Authors and Affiliations

  • Hyun Woo Choi
    • 1
    • 2
  • Jong Soo Kim
    • 1
  • Hyo Jin Jang
    • 1
    • 2
  • Sol Choi
    • 1
  • Jae-Hwan Kim
    • 4
  • Hans R. Schöler
    • 3
  • Jeong Tae Do
    • 1
  1. 1.Department of Animal Biotechnology, College of Animal Bioscience and TechnologyKonkuk UniversitySeoulRepublic of Korea
  2. 2.Department of Biomedical ScienceCHA UniversityGangnam-gu, SeoulRepublic of Korea
  3. 3.Department of Cell and Developmental BiologyMax Planck Institute for Molecular BiomedicineMünsterGermany
  4. 4.Department of Biomedical ScienceCHA UniversityGyeonggi-doRepublic of Korea

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