Influence of the vitrification solution on the angiogenic factors in vitrificated mouse ovarian tissue
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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.
Abstract
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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