Stepwise In-straw Dilution and Direct Transfer Using Open Pulled Straws (OPS) in the Mouse: A Potential Model for Field Manipulation of Vitrified Embryos

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概要

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• In the present study, mouse blastocysts were employed to investigate the feasibility and efficiency of stepwise in-straw dilution and direct transfer using the open pulled straw (OPS) method. In experiment I, the effects of various vitrification solutions (VS) on embryo survival were examined. After thawing, the expanded blastocyst rates (97.59% and 95.05%) and hatching rates (80.48% and 78.95%) achieved in the EDFS30 [15% ethylene glycol (EG), 15% dimethyl sulfoxide (DMSO), Ficoll, and sucrose] and EFS40 [40% EG, Ficoll, and sucrose] groups were no different from those (96.15% and 83.33%) of the control group. However, the rates in the EFS30 [30% EG, Ficoll, and sucrose] (87.80% and 55.43%) and EDFS40 [20% EG, 20% DMSO, Ficoll, and sucrose] (95.69% and 70.97%) groups were significantly lower than those (96.15% and 83.33%) of the control group (P<0.05). In the experiment II, the effects of the volume of VS in the OPS on the survival of embryos after in-straw thawing were investigated. When the length of the VS in the column was less than 1 cm, the in vitro viability of embryos thawed by stepwise in-straw dilution was no different among the experimental and control groups. The embryos could be successfully thawed by immersing the OPS in 0.5 M sucrose for 3 min and then 0.25M sucrose for 2 min. In experiment III, the effect of immersion time of the OPS in diluent (PBS) on the viability of vitrified embryos was investigated. After in-straw thawing, OPSs were immersed immediately in 1 ml PBS for 0 to 30 min. When the immersion time of the OPSs in PBS was less than 12 min, in vitro development of the in-straw thawed embryos was no different from that of the controls. In experiment IV, in-straw thawed blastocysts were directly transferred to pseudopregnant mice to examine their in vivo developmental viability. The pregnancy (91.67%) and birth rates (42.42%) of embryos in-straw thawed and directly transferred were no different from those of the unvitrified controls (90.90% and 40%) and embryos thawed by the conventional method (84.61% and 46.94%). These results demonstrate that mouse embryos vitrified with OPS could be successfully thawed by stepwise in-straw dilution and transferred directly to a recipient and that this method might be a model for field manipulation of vitrified embryos in farm animals.

• 日本繁殖生物学会の論文

著者

• Hou Yun-peng

  College Of Animal Science And Technology And State Key Laboratory For Agrobiotechnology China Agricu

• ZHU Shi-En

  Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, China Agricu

• Yang Qi-en

  Laboratory Of Animal Embryonic Biotechnology College Of Animal Science And Technology China Agricult

• Zhou Guang-bin

  College Of Animal Science And Technology And State Key Laboratory For Agrobiotechnology China Agricu

• Yang Zhong-qiang

  Laboratory Of Animal Embryonic Biotechnology College Of Animal Science And Technology China Agricult

• HOU Yun-Peng

  Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, China Agricu

• ZHOU Guang-Bin

  Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, China Agricu

• HOU Yun-Peng

  Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, China Agricultural University

• ZHU Shi-En

  Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, China Agricultural University

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• Tetraspanin CD9 in Bovine Oocytes and Its Role in Fertilization
• Piezo-assisted In Vitro Fertilization of Mouse Oocytes with Spermatozoa Retrieved from Epididymides Stored at 4 Degree C
• Stepwise In-straw Dilution and Direct Transfer Using Open Pulled Straws (OPS) in the Mouse : A Potential Model for Field Manipulation of Vitrified Embryos
• Stepwise In-straw Dilution and Direct Transfer Using Open Pulled Straws (OPS) in the Mouse: A Potential Model for Field Manipulation of Vitrified Embryos

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