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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 산부인과;대구마리아 산부인과;대구마리아 산부인과;대구마리아 산부인과;대구마리아 산부인과;대구마리아 산부인과;대구마리아 산부인과;서울마리아 산부인과 & 마리아 기초의학 연구소;서울마리아 산부인과 & 마리아 기초의학 연구소;서울마리아 산부인과 & 마리아 기초의학 연구소;대구대학교 축산학과;서울마리아 산부인과 & 마리아 기초의학 연구소;

김수희;이상원;이주희;강상민;오희정;이승민;이성구;윤혜균;윤산현;박세필;송해범;임진호;,

Study on the Vitrification of Human Blastocysts: II. Effect of Vitrification on the Implantation and the Pregnancy of Human Blastocysts

Kim, Su-Hee;Lee, Sang-Won;Lee, Ju-Hee;Kang, Sang-Min;Oh, Hee-Jeong;Lee, Seoung-Min;Lee, Seong-Goo;Yoon, Hye-Gyun;Yoon, San-Hyun;Park, Se-Pill;Song, Hai-Bum;Lim, Jin-Ho;

Taegu-Maria Obstetrics/Gynecology;Taegu-Maria Obstetrics/Gynecology;Taegu-Maria Obstetrics/Gynecology;Taegu-Maria Obstetrics/Gynecology;Taegu-Maria Obstetrics/Gynecology;Taegu-Maria Obstetrics/Gynecology;Taegu-Maria Obstetrics/Gynecology;Seoul-Maria Obstetrics/Gynecology & Maria Infertility Medical Institute;Seoul-Maria Obstetrics/Gynecology & Maria Infertility Medical Institute;Seoul-Maria Obstetrics/Gynecology & Maria Infertility Medical Institute;Department of Animal Science, Taegu University;Seoul-Maria Obstetrics/Gynecology & Maria Infertility Medical Institute;

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 derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts 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 ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ 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 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ 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 transfers. 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 slow 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 be chosen as a good method of human embryo freezing in IVF-ET programs.

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