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First published online July 13, 2006
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2005-0427v1
24/11/2549    most recent
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Submitted on August 31, 2005
Accepted on July 1, 2006

Stem Cell Genetics and Genomics

Dimethyl Sulfoxide (DMSO) Increases Expression of Dnmt3as and Affects Genome-wide DNA Methylation Profiles in Mouse Embryoid Body

Misa Iwatani 1, Kohta Ikegami 1, Yuliya Kremenska 1, Naka Hattori 1, Satoshi Tanaka 1, Shintaro Yagi 1, Kunio Shiota 1*

1 Laboratory of Cellular Biochemistry, Department of Animal Resource Sciences/Veterinary Medical Sciences, The University of Tokyo, Tokyo, Japan

* To whom correspondence should be addressed. E-mail: ashiota{at}mail.ecc.u-tokyo.ac.jp.


  Abstract

Dimethyl sulfoxide (DMSO), an amphipathic molecule, is widely used not only as a solvent for water-insoluble substances, but also as a cryopreservant for various types of cells. Exposure to DMSO sometimes causes unexpected changes in cell fates. Because mammalian development and cellular differentiation are controlled epigenetically by DNA methylation and histone modifications, DMSO likely affects the epigenetic system. The effects of DMSO on transcription of three major DNA methyltransferases (Dnmts) and five well studied histone modification enzymes were examined in mouse embryonic stem cells (ES cells) and embryoid bodies (EBs) by RT-PCR. Addition of DMSO (0.02-1.0%) to EBs in culture induced an increase in Dnmt3a mRNA level with increasing dosage. Increased expression of two subtypes of Dnmt3a in protein level was confirmed by Western blotting. Southern blot analysis revealed that DMSO caused hypermethylation of two kinds of repetitive sequences in EBs. Furthermore, Restriction Landmark Genomic Scanning (RLGS), by which DNA methylation status can be analyzed on thousands of loci in genic regions, revealed that DMSO affected DNA methylation status at multiple loci, inducing hypomethylation as well as hypermethylation depending on the genomic loci. In conclusion, DMSO has an impact on the epigenetic profile: upregulation of Dnmt3as' expression and alteration of genome-wide DNA methylation profiles with phenotypic changes in EBs.

Key Words. Dimethyl Sulfoxide (DMSO), DNA methylation, Epigenetics, Differentiation




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[Abstract] [Full Text] [PDF]


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