Piezo-assisted In Vitro Fertilization of Mouse Oocytes with Spermatozoa Retrieved from Epididymides Stored at 4 Degree C

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概要

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• This study was performed to investigate the effect of partial zona pellucida incision by piezo micromanipulation (ZIP) on the in vitro fertilizing ability of stored mouse spermatozoa. The storage conditions were optimized by storing the mouse epididymides at 4 C in mineral oil or in the mouse body for up to 4 days after death, and the retrieved spermatozoa were used to fertilize fresh oocytes. No significant difference was observed in fertilization rates between the treatments when epididymides were stored for up to 2 days, but the fertilization rates in mineral oil were higher (P<0.05) than those in the mouse body at 3 (41.4 vs. 16.2%) and 4 days (26.0 vs. 15.8%). Spermatozoa retrieved from epididymides stored in mineral oil were then used to fertilize fresh and vitrified oocytes with or without ZIP treatment. The fertilization rates of the ZIP fresh oocytes were higher than those of the zona-intact oocytes at each time point (1 to 4 days). After ZIP, the fertilization rates of spermatozoa stored for 1 and 2 days (91.2 and 86.6%, respectively) were similar (P>0.05) to that of fresh spermatozoa (91.9%). In regard to vitrified oocytes, the fertilization rates of zona-intact and ZIP oocytes using fresh spermatozoa were 46.7% and 84.7%, while the fertilization rates of vitrified ZIP oocytes using spermatozoa stored for 1 to 4 days ranged from 49.3 to 79.6%. When 2-cell embryos derived from ZIP fresh and vitrified oocytes inseminated with 2 day-stored spermatozoa were transferred into recipient females, 47.9 and 15.0% of the embryos developed to term, respectively. These results indicate that storing mouse epididymides at 4 C in mineral oil is more suitable than storage in the mouse body and that the ZIP technique improves the in vitro fertilizing ability of stored mouse spermatozoa in fresh oocytes and significantly increases the fertilization rate of vitrified oocytes with fresh spermatozoa.

• 2008-04-01

著者

• Zhu Shi-en

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

• Liu Ying

  Laboratory Animal Resource Center Tsukuba University

• Liu Ying

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

• Fan Zhi-qiang

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

• 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

• Zhou Guang-bin

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

• Meng Qing-gang

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

• ZHOU Guang-Bin

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

• LI Xiu-Wei

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

• MENG Qing-Gang

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

• WANG Yan-Ping

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

• HOU Yun-Peng

  National Key Laboratories for Agricultural Biotechnology, China Agricultural University

• Li Xiu-wei

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

• Wang Yan-ping

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

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