Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Vitrification of Mouse Embryos in Ethylene Glycol-based Solutions

에틸렌 글리콜 동결 보호제를 이용한 생쥐 배아의 유리화 동결 보존

  • Kim, Mi-Young (Department of Obstetrics and Gynecology, Chonnam National University Medical School) ;
  • Lee, Eun-Suk (Department of Obstetrics and Gynecology, Chonnam National University Medical School) ;
  • Lee, Seok-Won (Department of Obstetrics and Gynecology, Chonnam National University Medical School) ;
  • Lee, Yu-Il (Department of Obstetrics and Gynecology, Chonnam National University Medical School)
  • 김미영 (전남대학교 의과대학 산부인과학교실) ;
  • 이은숙 (전남대학교 의과대학 산부인과학교실) ;
  • 이석원 (전남대학교 의과대학 산부인과학교실) ;
  • 이여일 (전남대학교 의과대학 산부인과학교실)
  • Published : 2005.06.30
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Abstract

Objective: This study was conducted to find an optimal condition for the vitrification of mouse morulae and expanded blastocysts. Materials and Methods: Mouse embryos were obtained at 2-cell stage and cultured to morula and expanded blastocyst stage in Human Tubal Fluid (HTF) medium supplemented with 10% Serum Substitute Supplement (SSS). The vitrification solutions used were EFS30, EFS35 and EFS40 that contains 30%, 35% and 40% ethylene glycol, respectively, with 18% ficoll and 0.5 M sucrose diluted in Dulbecco's phosphate-buffered saline (DPBS) medium supplemented with 10% SSS. The vitrification procedure was performed in EFS solution with three steps, followed by thawing in 6 steps with 0.5 M sucrose, and then survival and hatching-hatched rate per embryos recovered were compared among six groups. Results: After 24 h culture in different vitrification and thawing solution, the survival rate of morula embryos was 94.1%, 85.4% and 59.7% for EFS30, EFS35 and EFS40 group, respectively. Hatching rate of morula embryos after 72 h culture was 30.6%, 25% and 11.3% for EFS30, EFS35 and EFS40 group, respectively. The survival rate of expanded blastocyst embryos after 24 h culture was 90.4%, 98.5% and 100% for EFS30, EFS35 and EFS40 group, respectively. Hatching rate of expanded blastocyst embryos after 48 h culture was 46.2%, 57.6% and 64.3% for EFS30, EFS35 and EFS40 group, respectively. Conclusion: The EFS30 solution was the best for vitrification of mouse morulae. The EFS40 solution was the best for vitrification of mouse expanded blastocysts. The mouse expanded blastocyst was better than mouse morula for vitrification of mouse embryos.

Keywords

References

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