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First published online January 10, 2008
Stem Cells Vol. 26 No. 3 March 2008, pp. 783 -788
doi:10.1634/stemcells.2007-0907; www.StemCells.com
© 2008 AlphaMed Press

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STEM CELL GENOMICS AND PROTEOMICS

Nuclear Transfer Alters the DNA Methylation Status of Specific Genes in Fertilized and Parthenogenetically Activated Mouse Embryonic Stem Cells

Takafusa Hikichia, Takashi Kohdab, Sayaka Wakayamaa, Fumitoshi Ishinob, Teruhiko Wakayamaa

aLaboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe Institute, Kobe, Japan;
bDepartment of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan

Key Words. Parthenogenesis • Nuclear transfer • Reprogramming • Embryonic stem cells

Correspondence: Takafusa Hikichi, Ph.D., 2-2-3 Minatojima-minamimachi Chuo-ku, Kobe 650-0047, Japan. Telephone: 81-78-306-3049; Fax: 81-78-306-3095; e-mail: hikichi{at}cdb.riken.jp

Received October 28, 2007; accepted for publication December 27, 2007.
First published online in STEM CELLS EXPRESS   January 10, 2008.



Recent cloning technology has been demonstrated successfully using nuclear transfer (NT) techniques to generate embryonic stem (ES) cells. Mice can be cloned from adult somatic cells or ES cells by NT, and such cloned embryos can be used to establish new NT-ES cell lines. However, ES cells derived from parthenogenetic embryos show epigenetic disorders and low potential for normal differentiation unless used to produce subsequent generations of NT-ES lines. Thus, enucleated oocytes can initialize epigenetic modification, but the extent and efficacy of this remain unclear. In this study, our goal was to clarify why the contribution rate of ES cells derived from parthenogenetic embryos (pES) cells appears to improve after NT. We compared gene expression profiles between pES and NT-pES cell lines using DNA microarray analysis and allele-specific DNA methylation analysis. Although changes in expression level were observed for 4% of 34,967 genes, only 81 (0.2%) showed common changes across multiple cell lines. In particular, the expression level of a paternally expressed gene, U2af1-rs1, was significantly increased in all NT-pES cell lines investigated. The methylation status at the upstream differentially methylated region of U2af1-rs1 was also changed significantly after NT. This was observed in NT-pES cells, but also in conventionally produced NT-ES cells, which has never been reported previously. These results suggest that NT affects the epigenetic status of a few gene regions in common and that a change in the methylation status of U2af1-rs1 could be used as a genetic marker to investigate the effects of NT.

Disclosure of potential conflicts of interest is found at the end of this article.






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