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KMID : 0358420130560060382
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Korean Journal of Obstetrics and Gynecology
2013 Volume.56 No. 6 p.382 ~ p.388
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Influence of the vitrification solution on the angiogenic factors in vitrificated mouse ovarian tissue
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Choi Won-Jun
Lee Ji-Hye
Park Mi-Hyun
Choi In-Young
Park Ji-Kwon
Shin Jeong-Kyu
Lee Soon-Ae
Paik Won-Young
Lee Jong-Hak
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Abstract
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Objective:To investigate the effect of the dimethyl sulfoxide (DMSO) and EFS-40 during vitrification on the expression of angiogenic factors in vitrified mouse ovarian tissue.
Methods:The ovarian tissues were obtained from 5 or 6 weeks aged ICR mouse. Ovarian tissues were divided into four groups: ovarian tissue without cryopreservation (control, group I), ovarian tissue vitrified with 15% DMSO (group II), ovarian tissue vitrified with EFS-40 (group III), and ovarian tissue slowly frozen with 10% DMSO (group IV). Thawing was carried out at room temperature. Levels of messenger RNA (mRNA) and protein for vascular endothelial growth factor-A (VEGF-A) and angiopoietin-2 (Angpt-2) were checked in ovarian tissues of four groups recovered on day 7 after cryopreservation. Reverse transcription-polymerase chain reaction and Western blot analysis were used to identify the levels of angiogenic factors in mouse ovarian tissues.
Results:Levels of mRNA and protein for VEGF-A and Angpt-2 were significantly decreased in cryopreserved group (group II, III and IV) than control group (group I) (P< 0.05). The significant differences of levels of mRNA and protein for VEGF-A and Angpt-2 between cryopreservation methods were observed (P< 0.05). Group III showed highest expression of mRNA and protein for VEFG-A and Angpt-2 than other cryopreservation groups (P< 0.05).
Conclusion:These findings suggest that EFS-40 is more efficient vitrification solution for preservation of angiogenic factors than 15% DMSO during vitrification of mouse ovarian tissue. Future studies should investigate to improve the vitrification solution for ovarian tissue vitrification.
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KEYWORD
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Angiopoietin-2, Ovary, Vascular endothelial growth factor A, Vitrification
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