Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein |
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Authors: | Atsushi IDETA Yoshito AOYAGI Kanami TSUCHIYA Yuuki NAKAMURA Kou HAYAMA Atsushi SHIRASAWA Kenichiro SAKAGUCHI Naomi TOMINAGA Yoshiyuki NISHIMIYA Sakae TSUDA |
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Institution: | 1)Research and Development Group, Zen-noh Embryo Transfer (ET) Center, Hokkaido 080–1407, Japan;2)Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Hokkaido 062–8517, Japan |
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Abstract: | Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming. |
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Keywords: | Antifreeze protein Bovine embryo Nonfreezing preservation |
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