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BOR - Papers in Press, published online ahead of print January 31, 2007.
Biol Reprod 2007, 10.1095/biolreprod.106.052779
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BIOLOGY OF REPRODUCTION 76, 900–905 (2007)
DOI: 10.1095/biolreprod.106.052779
© 2007 by the Society for the Study of Reproduction, Inc.

Production of Live Piglets Following Cryopreservation of Embryos Derived from In Vitro-Matured Oocytes1

Hiroshi Nagashima 2, Katsumi Hiruma , Hitoshi Saito , Ryo Tomii , Satoshi Ueno , Naoki Nakayama , Hitomi Matsunari , and Mayuko Kurome 

Laboratory of Developmental Engineering, Department of Life Science, School of Agriculture, Meiji University, Tama, Kawasaki 214-8571, Japan

ABSTRACT

We have successfully produced healthy piglets following cryopreservation of embryos derived from oocytes matured and fertilized in vitro. The appropriate timing of cryopreservation pretreatment (removal of cytoplasmic lipid droplets [delipation] and vitrification) was initially determined using parthenogenetic embryos derived from in vitro-matured (IVM) oocytes. Viable embryos were obtained at the highest rate when embryos were delipated at the four- to eight-cell stages (Day 2 of embryo culture) and were vitrified approximately 15 h later (Day 3) by means of the minimum volume cooling method. After cryopreservation of embryos derived from oocytes matured and fertilized in vitro under the most appropriate conditions, 401 embryos were transferred to five recipient gilts, and the recipients all became pregnant. At autopsy of one of the recipients, which had received 47 embryos, eight fetuses (17.0%) were found. Three recipients each gave birth to two to four piglets (1.4%–6.0%). These results demonstrate that normal offspring can be produced from vitrified porcine embryos derived from IVM oocytes by a strategic combination of delipation and vitrification at the early cleavage stages. This approach has great potential in the reproduction of micromanipulated porcine embryos, such as cloned and sperm-injected embryos, produced from IVM oocytes.

developmental biology,, embryo, in vitro fertilization

FOOTNOTES

1Supported by the 21st Century Green Frontier Research Project, National Institute of Agrobiological Science, Japan; the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN); and Biotechnology Venture Research and Development Base Maintenance Enterprise Project for Private Universities, Ministry of Education, Culture, Sports, Science and Technology, Japan, 2002–2006.

Correspondence: 2Hiroshi Nagashima, Laboratory of Developmental Engineering, Department of Life Science, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki, Kanagawa 214-8571, Japan. FAX: 81 44 934 7824; e-mail: hnagas{at}isc.meiji.ac.jp






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Copyright © 2007 by the Society for the Study of Reproduction.