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Sucrose affecting successful transplantation of vitrified-thawed mouse ovarian tissues.

ABSTRACT: PURPOSE: The aim of this experiment is to detect effects of varying levels of sucrose on vitrified ovarian tissues. METHODS: Ovarian tissues of mice were vitrified-thawed. Mice were randomly assigned to the fresh control group and experimental groups. According to different concentration of sucrose in vitrification solution, the experimental groups were randomly divided into Group I (0.2 M sucrose), Group II (0.4 M sucrose), Group III (0.8 M sucrose) and Group IV (1.6 M sucrose). Cytology was followed throughout the oophorectomy and transplantation period. Hormone levels and density of follicle were measured 1 month after transplantation. RESULTS: The number of days before the resumption of estrous cycles in control group was significantly smaller than those in all of experimental groups. The serum estradiol levels of mice and the follicular density of ovarian grafts in control group were significantly higher than those in all of experimental groups. In addition, the number of days before the resumption of estrous cycles in Group II and Group III were smaller than those in Group I and Group IV. The serum estradiol levels of mice and the follicular density of ovarian grafts in Group II and Group III were significantly higher than those in Group I and Group IV. However, no difference was observed in the number of days before the resumption of estrous cycles and the serum estradiol levels between Group II and Group III. A similar follicular density was also observed in Group II and Group III. CONCLUSION: Sucrose concentration of 0.4 M or 0.8 M in cryoprotective media is suitable for vitrifying mouse ovarian tissues.

SUBMITTER: Zhang JM

PROVIDER: S-EPMC2654937 | biostudies-other | 2009 Mar

REPOSITORIES: biostudies-other

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Project description:BackgroundAfter ovarian tissue transplantation, ischemia-reperfusion injury and free radicals cause follicle depletion and apoptosis. Therefore, the use of antioxidants to reduce the production of free radicals is an important method to address the consequences of ischemia-reperfusion injury. Resveratrol is a natural active polyphenol compound with anti-inflammatory, antitumor, strong antioxidant and anti-free radical properties. The aim of this study was to investigate whether resveratrol could improve the effect of autologous ovarian transplantation after cryopreserve-thawn mouse ovarian tissue.MethodsWhole-ovary vitrification and autotransplantation models were used to investigate the effects of resveratrol. Six-week-old female mice from the Institute of Cancer Research (ICR) were subjected to vitrification. All ovaries were preserved in liquid nitrogen for 1 week before being thawed. After thawing, ovarian tissues were autotransplanted in the bilateral kidney capsules. Mice (n = 72) were randomly divided into four groups to determine the optimal concentration of resveratrol (experiment I). Treatments were given as follows: saline, 5 mg/kg resveratrol, 15 mg/kg resveratrol and 45 mg/kg resveratrol, which were administered orally for one week. Grafted ovaries were collected for analysis on days 3, 7, and 21 after transplantation. Ovarian follicle morphology was assessed by hematoxylin and eosin staining. Serum FSH and E2 levels were measured to estimate the transplanted ovarian reserve and endocrine function. Other mice were randomly divided into two groups-saline and 45 mg/kg resveratrol to further evaluate the effect of resveratrol and explore the mechanisms underlying this effect (experiment II). Ovarian follicle apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assays. Immunohistochemistry, qRT-PCR and western blotting (MDA, SOD, NF-κB, IL-6 and SIRT1) were used to explore the mechanisms of resveratrol. Moreover, oocytes derived from autotransplanted ovaries at 21 days were cultured and fertilized in vitro.ResultsThe proportions of morphologically normal (G1) follicles at 3, 7 and 21 days were significantly higher in the 45 mg/kg resveratrol group than in the saline group. The TUNEL-stained follicles (%) at 7 days were significantly decreased in the 45 mg/kg resveratrol group compared with the saline group. Western blot analysis revealed that SOD2 and SIRT1 levels were significantly higher in the 45 mg/kg resveratrol group than in the saline group at day 7 and that MDA and NF-κB levels were lower in the saline group on day 3. Likewise, IL-6 was lower in the saline group on day 7. These results are basically consistent with the qRT-PCR results. In addition, the mean number of retrieved oocytes and fertilization and cleavage were significantly increased in the 45 mg/kg resveratrol group compared with the saline group.ConclusionsAdministration of resveratrol could improve the quality of cryopreserved mouse ovarian tissue after transplantation and the embryo outcome, through anti-inflammatory and antioxidative mechanisms.

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Sucrose affecting successful transplantation of vitrified-thawed mouse ovarian tissues.

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