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Production of somatic cell nuclear transfer embryos using in vitro-grown and in vitro-matured oocytes in rabbits

Published online by Cambridge University Press:  26 March 2014

Hironobu Sugimoto
Affiliation:
Taniguchi Hospital, 1–5-20 Onishi, Izumisano, Osaka 598-0043, Japan. Division of Biological Science, Graduate School of Biology-oriented Science and Technology, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan.
Yuta Kida
Affiliation:
Division of Biological Science, Graduate School of Biology-oriented Science and Technology, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan.
Noriyoshi Oh
Affiliation:
Division of Biological Science, Graduate School of Biology-oriented Science and Technology, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan.
Kensaku Kitada
Affiliation:
Kitayama Labes Co. Ltd., Ina Research Laboratory, Ina, Nagano 396-0021, Japan.
Kazuya Matsumoto
Affiliation:
Division of Biological Science, Graduate School of Biology-oriented Science and Technology, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan.
Kazuhiro Saeki
Affiliation:
Division of Biological Science, Graduate School of Biology-oriented Science and Technology, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan.
Takeshi Taniguchi
Affiliation:
Taniguchi Hospital, 1–5-20 Onishi, Izumisano, Osaka 598-0043, Japan.
Yoshihiko Hosoi*
Affiliation:
Department of Genetic Development, Kinki University, Wakayama 649-6493, Japan. Division of Biological Science, Graduate School of Biology-oriented Science and Technology, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan.
*
All correspondence to: Y. Hosoi. Department of Genetic Development, Kinki University, Wakayama 649-6493, Japan. Tel: +81 736 77 3888. Fax: +81 736 77 4754. e-mail: hosoi@waka.kindai.ac.jp

Summary

We examined growing oocytes collected from follicles remaining in superovulated rabbit ovaries, that were grown (in vitro growth, IVG) and matured (in vitro maturation, IVM) in vitro. We produced somatic cell nuclear transfer (SCNT) embryos using the mature oocytes and examined whether these embryos have the ability to develop to the blastocyst stage. In addition, we examined the effects of trichostatin A (TSA), a histone deacetylase inhibitor (HDACi), on the developmental competence of SCNT embryos derived from IVG–IVM oocytes. After growth for 7 days and maturation for 14–16 h in vitro, the growing oocytes reached the metaphase II stage (51.4%). After SCNT, these reconstructed embryos reached the blastocyst stage (20%). Furthermore, the rate of development to the blastocyst stage and the number of cells in the blastocysts in SCNT embryos derived from IVG–IVM oocytes were significantly higher for TSA-treated embryos compared with TSA-untreated embryos (40.6 versus 21.4% and 353.1 ± 59.1 versus 202.5 ± 54.6, P < 0.05). These results indicate that rabbit SCNT embryos using IVG–IVM oocytes have the developmental competence to reach the blastocyst stage.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2014 

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