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Development and spindle formation in rat somatic cell nuclear transfer (SCNT) embryos in vitro using porcine recipient oocytes

Published online by Cambridge University Press:  01 August 2009

Atsushi Sugawara ,
Satoshi Sugimura ,
Yumi Hoshino  and
Eimei Sato
Atsushi Sugawara*
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Sendai 981–8555, Japan.
Satoshi Sugimura
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1–1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan.
Yumi Hoshino
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1–1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan.
Eimei Sato
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1–1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai 981–8555, Japan.
*
All correspondence to: Atsushi Sugawara. Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Sendai 981–8555, Japan. Tel:/Fax: +81 22 717 8687. e-mail: [email protected]
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Summary

Cloning that uses somatic cell nuclear transfer (SCNT) technology with gene targeting could be a potential alternative approach to obtain valuable rat models. In the present study, we determined the developmental competence of rat SCNT embryos constructed using murine and porcine oocytes at metaphase II (MII). Further, we assessed the effects of certain factors, such as: (i) the donor cell type (fetal fibroblasts or cumulus cells); and (ii) premature chromosome condensation (PCC) with normal spindle formation, on the developmental competence of rat interspecies SCNT (iSCNT) embryos. iSCNT embryos that had been constructed using porcine oocytes developed to the blastocyst stage, while those embryos made using murine MII oocytes did not. Rat iSCNT embryos constructed with green fluorescent protein (GFP)-expressing fetal fibroblasts injected into porcine oocytes showed considerable PCC with a normal bipolar spindle formation. The total cell number of iSCNT blastocyst derived from GFP-expressing fetal fibroblasts was higher than the number derived from cumulus cells. In addition, these embryos expressed GFP at the blastocyst stage. This paper is the first report to show that rat SCNT embryos constructed using porcine MII oocytes have the potential to develop to the blastocyst stage in vitro. Thus the iSCNT technique, when performed using porcine MII oocytes, could provide a new bioassay system for the evaluatation of the developmental competence of rat somatic cells.

Keywords

Donor cell typesPorcine oocytesPremature chromosome condensationRat iSCNT
Type
Research Article
Information
Zygote , Volume 17 , Issue 3 , August 2009 , pp. 195 - 202
Copyright
Copyright © Cambridge University Press 2009

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