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Hum. Reprod. Advance Access originally published online on July 3, 2007
Human Reproduction 2007 22(9):2509-2514; doi:10.1093/humrep/dem206
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Effect of using slush nitrogen (SN2) on development of microsurgically manipulated vitrified/warmed mouse embryos

Dong Ryul Lee1,3, Yun Hee Yang1, Jin Hee Eum1, Jin Seong Seo1, Jung Jae Ko1, Hyung Min Chung1,2 and Tae Ki Yoon1

1 Fertility Center of CHA General Hospital, CHA Research Institute, Pochon CHA University, 606-5 Yeoksam-dong, Gangnam-gu, Seoul 135-081, Republic of Korea 2 Chabiotech Co. Ltd, Seoul 135-081, Republic of Korea

3 Correspondence address. Tel: +82-2-3468-3422; Fax: +82-2-501-8704; E-mail: drleedr{at}cha.ac.kr

BACKGROUND: This study evaluated the effect of vitrification using slush nitrogen (SN2) on cryopreservation of micromanipulated mouse embryos.

METHODS: The zona pellucida of 4-cell embryos was either left intact or dissected or dissected with biopsy of an intact blastomere. In a second study, a blastomere was destroyed and either removed (removed group) or not removed (remained group) prior to vitrification/freezing. The micromanipulated embryos were equilibrated and loaded into an open pulled straw (OPS), and plunged into liquid nitrogen (LN2) or SN2.

RESULTS: When using LN2 vitrification, recovery and blastocyst formation rates of embryos were lower for zona pellucida-opened and/or blastomere-biopsied embryos compared with zona pellucida-intact embryos. Using SN2 for vitrification resulted in increased survival and development of vitrified/warmed embryos in both the zona pellucida-opened and blastomere-biopsy groups. Similar results were observed when using embryos with a destroyed blastomere either removed or left remaining before vitrification. However, the number of total and apoptotic cells were similar for both LN2 and SN2. In addition, using SN2 increased the rate of intact recovery and blastocyst formation in warmed hemi-8-cell embryos derived from the same embryo.

CONCLUSIONS: These results suggest that vitrification using SN2 is useful in cryopreservation of micromanipulated embryos obtained from a variety of programs, including assisted hatching, preimplantation genetic diagnosis and nuclear transfer.

Key words: vitrification/slush nitrogen (SN2)/micromanipulated embryos/mouse embryo/embryonic development

Submitted on April 2, 2007; resubmitted on June 3, 2007; accepted on June 13, 2007.


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