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In vitro development of mouse somatic nuclear transfer embryos: effects of donor cell passages and electrofusion

Published online by Cambridge University Press:  01 August 2008

Gang Zhang ,
Qing-Yuan Sun  and
Da-Yuan Chen
Gang Zhang
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100080, P. R. China. Department of Cell and Systems Biology, University of Toronto, Toronto, Ontario, Canada.
Qing-Yuan Sun
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100080, P. R. China.
Da-Yuan Chen*
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, P. R. China. State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100080, P. R. China.
*
All correspondence to: Da-Yuan Chen. State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, P. R. China. Tel: +86 010 64807052. Fax: +86 010 64807090. e-mail: chendy@ioz.ac.cn.
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Summary

In this study, C57BL/6 adult male mouse ear fibroblast cells and Kunming mouse M2 oocytes were used as donors and recipients, respectively, to investigate the effect of passage number on donor cells and electrofusion times on the in vitro development of nuclear transfer (NT) embryos. The results demonstrated firstly that when the ear fibroblast cells from either 2–4, 5–7 or 8–10 passages were used as donors, respectively, to produce NT embryos, the number of passages undergone by the donor cells had no significant effect on the in vitro development of NT embryos. The developmental rates for morula/blastocyst were 15.2, 13.3 and 14.0%, respectively, which were not significantly difference (p > 0.05). Secondly, when the NT embryos were electrofused, there was no significant difference between the fusion ratio for the first electrofusion and the second electrofusion (p > 0.05). The developmental rates of the 2-cell and 4-cell stages that had undergone only one electrofusion, however, were significantly higher than those that had had two electrofusions (65.7% compared with 18.4% and 36.4% compared with 6.1%; p < 0.01), furthermore the NT embryos with two electrofusions could not develop beyond the 4-cell stage. This study suggests that this protocol might be an alternative method for mouse somatic cloning, even though electrofusion can exert negative effects on the development of NT embryos.

Keywords

Ear fibroblast cellsElectrofusionMouseNuclear transfer
Type
Research Article
Information
Zygote , Volume 16 , Issue 3 , August 2008 , pp. 223 - 227
Copyright
Copyright © Cambridge University Press 2008

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