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Effect of volume of oocyte cytoplasm on embryo development after parthenogenetic activation, intracytoplasmic sperm injection, or somatic cell nuclear transfer

Published online by Cambridge University Press:  01 August 2008

Wakayama Sayaka
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Kishigami Satoshi
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Nguyen Van Thuan
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Ohta Hiroshi
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Hikichi Takafusa
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Mizutani Eiji
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Bui Hong Thuy
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
Miyake Masashi
Affiliation:
Department of Life Science, Graduate School of Science and Technology, Kobe University, Kobe 657–8501, Japan.
Wakayama Teruhiko*
Affiliation:
Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, 2–2–3 Minatojima-minamimachi Chuo-ku, Kobe 650–0047, Japan. Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, Japan.
*
All correspondence to: Wakayama Teruhiko. Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN Kobe, 2–2–3 Minatojima-minamimachi Chuo-ku, Kobe 650–0047, Japan. Tel: +81 78 306 3049. Fax: +81 78 306 0101. e-mail [email protected]

Summary

Animal cloning methods are now well described and are becoming routine. Yet, the frequency at which live cloned offspring are produced remains below 5%, irrespective of the nuclear donor species or cell type. One possible explanation is that the reprogramming factor(s) of each oocyte is insufficient or not properly adapted for the receipt of a somatic cell nucleus, because it is naturally prepared only for the receipt of a gamete. Here, we have increased the oocyte volume by oocyte fusion and examined its subsequent development. We constructed oocytes with volumes two to nine times greater than the normal volume by the electrofusion or mechanical fusion of intact and enucleated oocytes. We examined their in vitro and in vivo developmental potential after parthenogenetic activation, intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). When the fused oocytes were activated parthenogenetically, most developed to morulae or blastocysts, regardless of their original size. Diploid fused oocytes were fertilized by ICSI and developed normally and after embryo transfer, we obtained 12 (4–15%) healthy and fertile offspring. However, enucleated fused oocytes could not support the development of mice cloned by SCNT. These results suggest that double fused oocytes have normal potential for development after fertilization, but oocytes with extra cytoplasm do not have enhanced reprogramming potential.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2008

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