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Effects of sucrose treatment on the development of mouse nuclear transfer embryos with morula blastomeres as donors

Published online by Cambridge University Press:  01 February 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, 100101, China. Department of Cell and Systems Biology, University of Toronto, 25 Harbord Street, RW618, Toronto, Ontario, M5S 3G8, Canada.
Qing-Yuan Sun
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, China.
Da-Yuan Chen*
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing, 100101, China.
*
1All correspondence to: Da-Yuan Chen. State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing, 100101, China. Tel: +86 010 64807052. Fax: +86 010 64807316. e-mail: chendy@ioz.ac.cn
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Summary

In this study, nuclear transfer (NT) embryos were produced by using C57Bl/6 mouse morula blastomeres and Kunming mouse metaphase II (MII) oocytes as donors and recipients, respectively, to investigate the effects of sucrose treatment of MII oocytes with different concentrations on the manipulation time of NT, electrofusion and the in vitro and in vivo development of reconstructed embryos. The results demonstrated that: (i) when the oocytes were enucleated with 1, 2 and 3% sucrose treatment, respectively, the enucleating rates were not affected by the different sucrose concentrations, but the manipulation time had significant difference and the mean nuclear transfer manipulation times of every oocyte were 180 ± 10 s, 130 ± 10 s and 120 ± 10 s, respectively; (ii) different sucrose concentrations had no significant effects on the fusion rate and the in vitro developmental potential of the NT embryos (p > 0.05). Furthermore, 59 embryos were transplanted into the oviducts of two recipients. In the end, three dead full-term developed fetuses were obtained on 21 days post coitus (dpc). These results suggested that the mouse MII oocytes enucleated via sucrose treatment might be an alternative source for mouse cloning and could support the embryonic NT embryos developed to term in vivo.

Keywords

Morula blastomeresMouseNuclear transferSucrose treatment
Type
Research Article
Information
Zygote , Volume 16 , Issue 1 , February 2008 , pp. 15 - 19
Copyright
Copyright © Cambridge University Press 2008

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