Hum. Reprod. Advance Access originally published online on October 7, 2004
Human Reproduction 2005 20(1):122-128; doi:10.1093/humrep/deh540
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Successful pregnancy following blastocyst cryopreservation using super-cooling ultra-rapid vitrification
1 Division of Infertility Clinic, Lee Women's Hospital, 2 Institute of Biochemistry, Chung-Shan Medical University and 5 Department of Obstetrics and Gynecology, Chung-Shan Medical University Hospital, Taichung, 3 Department of Biotechnology, Chungtai Institute of Health Science and Technology, 4 Department of Obstetrics and Gynecology, College of Medicine and the Hospital, National Taiwan University, Taipe, and 6 Department of Medicine, China Medical University, Taiwan
7 To whom correspondence should be addressed at: Lee Women's Hospital, 263 Pei-Tun Road, Taichung 406, Taiwan. Email: msleephd{at}giga.net.tw
BACKGROUND: Blastocysts were cryopreserved by a new two-step ultra-rapid cooling in super-cooled liquid nitrogen (–205°C). METHODS: There were 308 mouse blastocysts collected from fertile B6CBF1 mice and 249 human blastocysts collected from 51 couples treated with IVF. The blastocysts were super-cooled by a Vit-MasterTM and cryoloops after treatment in 50 and 100% vitrification solution (VS) for 2 min and 30 s, respectively. The 100% VS was composed of 20% ethylene glycol, 20% dimethylsulphoxide and 0.5 mol/l sucrose in human tubular fluid medium with 20% human serum albumin. The embryos were warmed after treatment in 0.25 and 0.125 mol/l sucrose for 2 and 3 min, respectively. The survival of embryos was observed after re-swell. RESULTS: The survival rate (SR) and hatching rate (HR) of mouse blastocysts in the super-cooled, the cryosolution-treated and control groups were not significantly different (SR, 87, 95.5 and 100%; HR, 50, 33 and 44.6%, respectively; P>0.05). After 96 super-cooled human blastocysts were warmed, 60 survival blastocysts were transferred into 13 patients. The successful SR and pregnancy rate (PR) for the super-cooled blastocyst group were 77.1% (74 out of 96) and 53.8% (seven out of 13). CONCLUSION: The ultra-rapid vitrification of blastocysts with a successful SR and PR could be used to replace classical slow cooling.
Key words: blastocyst cryopreservation/super-cooling,ultra-rapid vitrification
* These authors contributed equally to this work.
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