Effects of Timing of Fusion and Culture Medium on the Development of Bovine Eggs Receiving Blastomeres from 8 to 32-cell Embryos In Vitro

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  • KATO Yoko
    Laboratory of Animal Reproduction, College of Agriculture, Kinki University
  • NITTA Ryohei
    Laboratory of Animal Reproduction, College of Agriculture, Kinki University
  • TAKANO Hiroshi
    Nara Prefectual Livestock Experimental Station
  • TSUNODA Yukio
    Laboratory of Animal Reproduction, College of Agriculture, Kinki University

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  • ウシ核移植卵の体外発生能に及ぼす融合時期ならびに培養液の影響
  • ウシ カク イショク ラン ノ タイガイ ハッセイノウ ニ オヨボス ユウゴウ

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Abstract

Effects of timing of fusion and culture medium on the development of bovine eggs receiving blastomeres from 8-to 32-cell embryos were examined in vitro. Donor embryos were produced from immature oocytes obtained from ovaries following in-vitro maturation, fertilization and culture for 3 to 4 days. Recipient oocytes were also used after in-vitro maturation for 22 hours in media supplemented with various agents. Electrofusion (750V/cm for 50μsec, DC pulses 2 times) was performed at 30 or 40 hours from maturational culture of recipient oocytes. Reconstituted eggs were co-cultured with follicular cells in TCM-199 medium supplemented with hormones (LH, FSH, estradiol) and/or SOD + thioredoxin, or in M16 medium replaced glucose with gultamine. The results obtained were as follows. Overall fusion rate was 31 to 57%. Reconstituted eggs fused at 40 hours developed to 8-cell stage onwards at the higher rate than those of fused at 30 hours (7 to 23% VS 7 to 32%). However, supplements of hormones, SOD + thioredoxin or gultamine did not improve the developmental ability of nuclear transfer eggs.

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