title: Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein.
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.
authors.affiliation: Research and Development Group, Zen-noh Embryo Transfer (ET) Center, Hokkaido 080-1407, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ideta
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Aoyagi
authors.forename: Kanami
authors.initials: K
authors.lastname: Tsuchiya
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Kou
authors.initials: K
authors.lastname: Hayama
authors.forename: Atsushi
authors.initials: A
authors.lastname: Shirasawa
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Sakaguchi
authors.forename: Naomi
authors.initials: N
authors.lastname: Tominaga
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Nishimiya
authors.forename: Sakae
authors.initials: S
authors.lastname: Tsuda
mesh_terms: D000818:Animals; D021343:Antifreeze Proteins, Type III / Q000737:chemistry*; D002417:Cattle; D002470:Cell Survival; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media; D046149:Embryo Culture Techniques; D029941:Fish Proteins / Q000737:chemistry; D008856:Microscopy, Fluorescence; D010473:Perciformes; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors
chemical_list: D021343:Antifreeze Proteins, Type III; D003470:Culture Media; D029941:Fish Proteins; D011994:Recombinant Proteins