title: Cryopreservation and In Vitro culture of Preimplantation Embryos in Djungarian Hamster (Phodopus sungorus).
abstract: Although embryo cryobanking was applied to Syrian golden and to Campbell's hamsters, no attempt has been made at freezing embryos in Djungarian hamsters. Four-cell stage embryos were flushed from the reproductive ducts of pregnant females before noon of the third-day post coitum and frozen in 0.25-ml straws according to standard procedures of slow cooling. A mixture of permeating (ethylene glycol) and non-permeating (sucrose) cryoprotectants was used. The thawing was performed by incubating at RT for 40 s followed by 40 s in a water bath at 30.0°C. Most (66.7%) of the non-frozen four-cell embryos developed up to the morula stage in rat one-cell embryo culture medium (R1ECM). The use of hamster embryo culture medium (HECM) yielded fewer morulas (18.2%) during the same 24-h period of culture. The rate of embryo's surviving the freezing-thawing procedures, as estimated by light microscopy, was 60.7-68.8%. After 24-h culturing in R1ECM, 64.7% of frozen-thawed four-cell embryos developed and all of them reached the morula stage. Supplementation of R1ECM with GM-CSF (2 ng/ml) improved the rate of Djungarian hamster frozen-thawed embryo development: 100% of the four-cell stage embryos developed, 50% of them achieved the morula stage, and 50% developed even further and reached the blastocyst stage within 24 h of culturing. This study reports the world's first successful transfer of frozen-thawed Djungarian hamster embryos yielding term pups. Taken together, the results of this study demonstrate the possibility of applying some key reproductive technologies, that is, embryo freezing/cryopreservation and in vitro culture, to Djungarian hamsters.
authors.affiliation: Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: E Yu
authors.initials: EY
authors.lastname: Brusentsev
authors.affiliation: Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: T O
authors.initials: TO
authors.lastname: Abramova
authors.affiliation: Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: I N
authors.initials: IN
authors.lastname: Rozhkova
authors.affiliation: Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: T N
authors.initials: TN
authors.lastname: Igonina
authors.affiliation: Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: V A
authors.initials: VA
authors.lastname: Naprimerov
authors.affiliation: Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, Moscow, Russia.
authors.forename: N Yu
authors.initials: NY
authors.lastname: Feoktistova
authors.affiliation: Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: S Ya
authors.initials: SY
authors.lastname: Amstislavsky
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D006224:Cricetinae; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003470:Culture Media; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000008:administration & dosage; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage; D008969:Molecular Sequence Data; D009028:Morula / Q000502:physiology; D017111:Phodopus / Q000196:embryology*; D011247:Pregnancy
chemical_list: D003470:Culture Media; D004815:Epidermal Growth Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor