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. 2017 Oct;19(3):492-505.
doi: 10.22074/cellj.2017.4513. Epub 2017 Aug 19.

Comparison of Allotransplantation of Fresh and Vitrified Mouse Ovaries to The Testicular Tissue under Influence of The Static Magnetic Field

Vida Sadat Kazemein Jasemi  1 , Firooz Samadi  1 , Hussein Eimani  2   3 , Saeed Hasani  1 , Rouhollah Fathi  2 , Abdolhossein Shahverdi  2
Affiliations

Comparison of Allotransplantation of Fresh and Vitrified Mouse Ovaries to The Testicular Tissue under Influence of The Static Magnetic Field

Vida Sadat Kazemein Jasemi et al. Cell J. 2017 Oct.

Abstract

Objectives: The aim of this study was to investigate the effects of static magnetic field (SMF) during transplantation of the ovarian tissue into the testis.

Materials and methods: In this experimental study, ovaries of 6- to 8-week-old female Naval Medical Research Institute (NMRI) mice were randomly divided into four groups: i. Fresh ovaries were immediately transplanted into the testicular tissue (FOT group), ii. Fresh ovaries were exposed to the SMF for 10 minutes and then transplanted into the testicular tissue (FOT+ group), iii. Vitrified-warmed ovaries were transplanted into the testicular tissue (VOT group), and iv. Vitrified-warmed ovaries were transplanted into the testicular tissue and the transplantation site was then exposed to the SMF for 10 minutes (VOT+ group).

Results: The lowest percentages of morphologically dead primordial follicles and the highest percentages of morphologically intact primordial follicles were seen in the FOT+ group (4.11% ± 2.88 and 41.26% ± 0.54, respectively). Although the lowest significant percentage of maturation, embryonic development and fertility was observed in the VOT group as compared to the other groups, the difference in the fertility rate was not significant between the VOT and VOT+ groups. Estrogen and progesterone concentrations were significantly higher in the FOT+ group than those of the control mice.

Conclusions: It is concluded that, exposure of the vitrified-warmed ovaries to SMF retains the structure of the graft similar to that of fresh ovaries.

Keywords: Apoptosis; Magnetic Field; Mice; Transplantation; Vitrification.

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Conflict of interest statement

There is no conflict of interest in this study.

Figures

Fig.1
Fig.1
Image of the static magnetic field generating system. Vitrified-warmed ovaries were transplanted into the testicular tissue and the transplantation site was then exposed to a magnetic field for 10 minutes (VOT+ group).
Fig.2
Fig.2
Morphology of mouse ovarian histological sections transplanted into the testis. A. Fresh ovaries were immediately transplanted into the testicular tissue (FOT group), B. Fresh ovaries were exposed to the magnetic field for 10 minutes and then transplanted into the testicular tissue (FOT+ group), C. Vitrified-warmed ovaries were transplanted into the testicular tissue (VOT group), and D. Vitrifiedwarmed ovaries were transplanted into the testicular tissue and the transplantation site was then exposed to a magnetic field for 10 minutes (VOT+ group). White arrow indicates preantral follicle. Yellow arrow indicates primary follicle. T indicates the host tissue (testis). Green arrow indicates the host tissue spermatozoa (scale bars: 10 μm).
Fig.3
Fig.3
Immunohistochemical analysis for blood vessels (using the marker CD31) in the ovarian tissue transplanted into the testis. The liver tissue was used as A. Positive and B. Negative controls for CD-31, C. Fresh ovaries were immediately transplanted into the testicular tissue (FOT group), D. Fresh ovaries were exposed to the magnetic field for 10 minutes and then transplanted into the testicular tissue (FOT+ group), E. Vitrified-warmed ovaries were transplanted into the testicular tissue (VOT group), and F. Vitrified-warmed ovaries were transplanted into the testicular tissue and the transplantation site was then exposed to a magnetic field for 10 minutes (VOT+ group). Black arrows indicate blood vessels (scale bars: 10 μm).
Fig.4
Fig.4
Immunohistochemical analysis for caspase-3 in the ovarian tissue transplanted into the testis. The thymus tissue was used as A. Positive, B. Negative controls for caspase-3, C. Fresh ovaries were immediately transplanted into the testicular tissue (FOT group), D. Fresh ovaries were exposed to the magnetic field for 10 minutes and then transplanted into the testicular tissue (FOT+ group), E. Vitrified-warmed ovaries were transplanted into the testicular tissue (VOT group), and F. Vitrified-warmed ovaries were transplanted into the testicular tissue and the transplantation site was then exposed to a magnetic field for 10 minutes (VOT+ group). Positive staining is shown as brown coloration of the cytoplasm of the cells, and white arrow indicates caspase-3 staining (scale bars: 10 μm).
Fig.5
Fig.5
In vitro maturation, fertilization and cleavage of mouse germinal vesicle (GV) oocytes obtained from the ovarian tissue transplanted into the testis. A. Mouse GV oocyte, B. Oocytes extruded a polar body after overnight in vitro maturation, C. Fertilised 2-pronuclear zygote, and D-F. Development of oocytes derived from in vitro culturing after fertilization (scale bars: 10 μm).
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