Solid-surface vitrification is an appropriate and convenient method for cryopreservation of isolated rat follicles

Reproductive Biology and Endocrinology

Research

Markus Montag¹  Yubin Li²  Jiang Bian²  Tao Li²  Weijie Xing²  Canquan Zhou²  Yanwen Xu² 

[1] Department of Gynecological Endocrinology and Reproductive Medicine, University Clinics Bonn, Sigmund-Freud-Str. 25, 53105, Bonn, Germany;Reproductive Medicine Center, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China;

关键词: Granulosa Cell;    Rough Endoplasmic Reticulum;    Alginate Hydrogel;    Perivitelline Space;    Vitrification Solution;   

DOI  :  10.1186/1477-7827-8-42

 received in 2009-12-30, accepted in 2010-05-11,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundCryopreservation of isolated follicles may be a potential option to restore fertility in young women with cancer, because it can prevent the risks of cancer transmission. Several freezing protocols are available, including slow-rate freezing, open-pulled straws vitrification (OPS) and solid-surface vitrification (SSV, a new freezing technique). The purpose of our study was to investigate the effects of these freezing procedures on viability, ultrastructure and developmental capacity of isolated rat follicles.MethodsIsolated follicles from female Sprague-Dawley rats were randomly assigned to SSV, OPS and slow-rate freezing groups for cryopreservation. Follicle viability assessment and ultrastructural examination were performed after thawing. In order to study the developmental capacity of thawed follicles, we performed in vitro culture with a three-dimensional (3D) system by alginate hydrogels.ResultsOur results showed that the totally viable rate of follicles vitrified by SSV (64.76%) was slightly higher than that of the OPS group (62.38%) and significantly higher than that of the slow-rate freezing group (52.65%; P < 0.05). The ultrastructural examination revealed that morphological alterations were relatively low in the SSV group compared to the OPS and slow-rate freezing groups. After in vitro culture within a 3D system using alginate hydrogels, we found the highest increase (28.90 ± 2.21 μm) in follicle diameter in follicles from the SSV group. The estradiol level in the SSV group was significantly higher than those in the OPS and slow-rate freezing groups at the end of a 72-hr culture period (P < 0.05).ConclusionsOur results suggest that the SSV method is an appropriate and convenient method for cryopreservation of isolated rat follicles compared with the conventional slow-rate freezing method and the OPS method.

【 授权许可】

Unknown   
© Xing et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.