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First published online May 11, 2006
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2005-0537v1
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Submitted on October 31, 2005
Accepted on May 3, 2006

Embryonic Stem Cells

Equivalency of Nuclear Transfer-Derived Embryonic Stem Cells to those Derived from Fertilized Mouse Blastocyst

S. Wakayama 1, M. L. Jakt 2, M. Suzuki 3, R. Araki 4, T. Hikichi 5, S. Kishigami 5, H. Ohta 5, N. Van Thuan 5, E. Mizutani 6, Y. Sakaide 5, S. Senda 3, S. Tanaka 3, M. Okada 2, M. Miyake 7, M. Abe 8, S. I. Nishikawa 2, K. Shiota 3, T. Wakayama 5*

1 Center for Developmental Biology, RIKEN , Kobe, Japan; Department of Life Science, Graduate School of Science and Technology, Kobe University, Kobe, Japan
2 Stem Cell Biology Laboratory, Center for Developmental Biology, RIKEN, Kobe, Japan
3 Cellular Chemistry, Animal Resource Sciences/Veterinary Medical Sciences, The University of Tokyo, Tokyo, Japan
4 Transcriptome Research Center, National Institute of Radiological Sciences (NIRS), Chiba-shi, Japan
5 Center for Developmental Biology, RIKEN, Kobe, Japan
6 Center for Developmental Biology, RIKEN, Kobe, Japan; Graduate School of Agricultural Science, Tohoku University, Sendai, Japan
7 Department of Life Science, Graduate School of Science and Technology, Kobe University, Kobe, Japan
8 Transcriptome Research Center, National Institute of Radiological Sciences (NIRS), Chiba-shi, Japan

* To whom correspondence should be addressed. E-mail: teru{at}cdb.riken.jp.


  Abstract

Therapeutic cloning, whereby nuclear transfer (NT) is used to generate embryonic stem cells (ESCs) from blastocysts, has been demonstrated successfully in mice and cattle. However, if NT-ESCs have abnormalities, such as those associated with the offspring produced by reproductive cloning, then their scientific and medical utilities might prove limited. To evaluate the characteristics of NT-ESCs, we established more than 150 NT-ESC lines from adult somatic cells of several mouse strains. Here, we show that these NT-ESCs were able to differentiate into all functional embryonic tissues in vivo. Moreover, they were identical to blastocyst-derived ESCs in terms of their expression of pluripotency markers, in the presence of tissue-dependent differentially DNA methylated regions, in DNA microarray profiles and in high-coverage gene expression profiling. Importantly, the NT procedure did not cause irreversible damage to the nuclei. These similarities of NT-ESCs and ESCs indicate that murine therapeutic cloning by somatic cell NT can provide a reliable model for preclinical stem cell research.

Key Words. nuclear transfer, cloning, embryonic stem, reprogramming




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