Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Development of Effective Cryopreservation Method for Mouse Oocytes

생쥐 난자의 효율적인 냉동보존 방법 확립을 위한 연구

  • Choi, Su-Jin (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women's Healthcare Center) ;
  • Kim, Soo-Kyung (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women's Healthcare Center) ;
  • Kim, Ji-Sun (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women's Healthcare Center) ;
  • Cho, Jae-Won (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women's Healthcare Center) ;
  • Jun, Jin-Hyun (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women's Healthcare Center) ;
  • Byun, Hye-Kyung (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital and Women's Healthcare Center)
  • 최수진 (삼성제일병원 생식생물학 및 불임 연구실) ;
  • 김수경 (삼성제일병원 생식생물학 및 불임 연구실) ;
  • 김지선 (삼성제일병원 생식생물학 및 불임 연구실) ;
  • 조재원 (삼성제일병원 생식생물학 및 불임 연구실) ;
  • 전진현 (삼성제일병원 생식생물학 및 불임 연구실) ;
  • 변혜경 (삼성제일병원 생식생물학 및 불임 연구실)
  • Published : 2004.03.30
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Abstract

Objective: The purpose of this study was to evaluate the efficacy and effect of various cryopreservation method on the survival and the cytoskeletal stability of metaphase II mouse oocyte. Methods: Mouse ovulated oocytes were collected and cryopreserved by a modified slow-freezing method with 1.5 M 1, 2-propanediol (PrOH)+0.1 M sucrose or by vitrification using cryo loop and EM grid with 40% ethylene glycol+0.6 M sucrose. Four hours after thawing, intact oocytes were fixed and stained with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-$\beta$-tubulin antibody to visualize spindle and propidium iodide (PI) to visualize chromosome. Spindle morphology was classified as follows: normal (barrel-shaped), slightly and absolute abnormal (multipolar or absent). Results: Survival rate of the frozen-thawed oocytes in vitrification group was significantly higher than that of slow-freezing group (62.7% vs. 24.4%, p<0.01). Vitrification with cryo loop showed significantly higher survival rate than that with EM grid (67.7% vs. 53.5%, p<0.05). On the other hand, proportion of normal spindle and chromosome configurations of the frozen-thawed oocytes between two vitrification group was not significantly different. Conclusion: For mouse ovulated oocytes, vitrification with cryo loop may be a preferable procedure compared to slow-freezing method. Further study should be needed to investigate developmental competency of frozen-thawed mouse oocytes.

Keywords

References

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