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Mona Sheikhi1,*, Kjell Hultenby2, Boel Niklasson1, Monalill Lundqvist1 and Outi Hovatta11Division of Obstetrics and Gynaecology, Department of Clinical, Science, Technology and Intervention, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden 2Division of Clinical Research Centre, Department of Laboratory Medicine, Karolinska Institutet, SE 141 86 Stockholm, Sweden *Correspondence address. Tel: +46-8-5858-0000; Fax: +46-8-58587250; E-mail: mona.sheikhi{at}karolinska.seReceived March 17, 2010. Revision received November 10, 2010. Accepted November 22, 2010. BACKGROUND Cancer therapy is one of many conditions which may diminish the ovarian reserve. Banking of human ovarian tissue has become an option for the preservation of female fertility. We have shown that vitrification is an excellent method to cryopreserve ovarian tissue. To carry out vitrification in a clinical setting, we have developed a clinical grade closed system to avoid direct contact of ovarian tissue with liquid nitrogen. METHODS Ovarian tissue was obtained by biopsy from 12 consenting women undergoing Caesarean section. Tissues were vitrified in cryotubes, using dimethyl sulphoxide, 1,2-propanediol, ethylene glycol and polyvinylpyrrolidon as cryoprotectants. Non-vitrified and warmed-vitrified tissue was compared by light and electron microscopic morphology of the follicles within the tissues. RESULTS We did not see any differences in the light or electron microscopic ultrastructure of oocytes between non-vitrified and vitrified tissues. No irreversible subcellular alterations in vitrified tissues were seen. CONCLUSIONS The ultrastructure of follicles within the vitrified human ovarian tissue was well preserved using cryotube in a closed vitrification system to avoid direct contact of liquid nitrogen. The system is compatible with the European tissue directive. © The Author 2011. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This ArticleHum. Reprod. (2011) 26 (3): 594-603. doi: 10.1093/humrep/deq357 First published online: January 8, 2011
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