Cloned Porcine Embryos can Maintain Developmental Ability after Cryopreservation at the Morula Stage

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  • NAKANO Kazuaki
    Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University
  • MATSUNARI Hitomi
    Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University
  • NAKAYAMA Naoki
    Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University
  • OGAWA Buko
    Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University
  • KUROME Mayuko
    Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University Institute of Molecular Animal Breeding and Biotechnology, Ludwig-Maximilians-University Munich
  • TAKAHASHI Masashi
    National Agricultural Research Center for Kyushu Okinawa Region
  • MATSUMOTO Mitsuhito
    National Agricultural Research Center for Kyushu Okinawa Region
  • MURAKAMI Hitoshi
    National Agricultural Research Center for Kyushu Okinawa Region
  • KAJI Yuji
    National Agricultural Research Center for Kyushu Okinawa Region
  • NAGASHIMA Hiroshi
    Laboratory of Developmental Engineering, Department of Life Sciences, School of Agriculture, Meiji University Meiji University International Cluster for Bio-Resource Research

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The aim of the present study was to clarify the overall efficiency of porcine somatic cell nuclear transfer (SCNT) by incorporating cryopreservation of the cloned embryos before transfer. The SCNT embryos reconstructed with preadipocytes and in vitro-matured (IVM) oocytes were cultured to harvest morula stage embryos; they were then subjected to delipation (removal of cytoplasmic lipid droplets) and vitrification. After warming and culture, the embryos developing to blastocysts were transferred to recipients to obtain cloned piglets. From 372 reconstructed embryos, 188 (50.5%) reached the morula stage and 117 (31.5%) developed to blastocysts after vitrification. Transfer of 98 (26.3%) morphologically normal blastocysts gave rise to 6 (1.6%) piglets, including 1 stillborn. The efficiency of the cloned piglet production was comparable with that obtained using SCNT embryos without cryopreservation (2.7%, 17/635). Here, we demonstrate that porcine somatic cell cloning can be performed without a significant reduction in efficiency even when the SCNT embryos are cryopreserved before transfer.<br>

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