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대한생식의학회지 제27권 제1호 2010년
인간 포배기 배아의 초자화 동결에 관한 연구: Ⅱ. 초자화 동결이 포배기 배아의 착상 및 임신에 미치는 영향
대구마리아 산부인과1, 서울마리아 산부인과 & 마리아 기초의학 연구소2, 대구대학교 축산학과3
김수희1, 이상원1, 이주희1, 강상민1, 오희정1, 이승민1, 이성구1, 윤혜균2, 윤산현2, 박세필2, 송해범3, 임진호2,
Study on the Vitrification of Human Blastocysts: Ⅱ. Effect of Vitrification on the Implantation and the Pregnancy of Human Blastocysts
Su Hee Kim1, Sang Won Lee1, Ju Hee Lee1, Sang Min Kang1, Hee Jeong Oh1, Seoung Min Lee1, Seong Goo Lee1, Hye Gyun Yoon2, San Hyun Yoon2, Se Pill Park2, Hai Bum Song3, Jin Ho Lim2
Taegu-Maria Obestetrics/Gynecology1, Seoul-Maria Obstetrics/Gynecology & Maria Infertility Medical Institute2, Department of Animal Science, Taegu University3, Korea
Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derived from IVF were cocultured with cumulus cells in YS medium containing 20% Hff for 5 days. Two or three of the best blastocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer (-2℃/min to -7℃, manual seeding at -7℃, -0.3℃/min to -38℃ and plunged into LN2). The blastocysis frozen by slow freezing were thawed at 36℃ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps(10% glycerol for 5 min 10% glycerol + 20% ethylene glycol for 5 in, 25% glycerol + 25% ethylene glycol and directly LN2 within 1 min). The blastocysts frozen by vitrification were thawed at 20℃ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfer. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the sow freezing (clinical pregnancy rate; 40.3%), ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will chosen as a good method of human embryo freezing in IVF-ET programs.
키워드 : Vitrification, slow freezing, Human blastocyst embryos
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