Document Type : Original Article


1 Department of Animal and Poultry Physiology, Faculty of Animal Sciences, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran

2 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

3 Department of Anatomy, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran

4 Department of Biology, Payame Noor University, Tehran, Iran


This study was designed to investigate the effects of applying 1 mT static magnetic field (SMF) during the vitrification process, on the viability of ovarian follicles after vitrification-warming and autotransplantation. The study was conducted in two phases. In the first phase, ovaries of female NMRI mice (6 to 8 weeks old) were randomly divided into three groups: 1- Freshly isolated ovaries fixed in Bouin solution (control group), 2- Ovaries vitrified-warmed without exposure to magnetic field (V1 group) and 3- Ovaries exposed to magnetic field during equilibration step of the vitrification process (V2 group). In the second phase, the vitrified (V1 and V2 groups) and fresh ovarian tissues were autografted into the back muscles of the mice from which the ovaries were extracted. In both phases, morphological aspects and molecular characteristics of active-apoptotic caspase-3 antibody were evaluated. Results indicated the lower percentages of morphologically intact primordial, primary and antral follicles in the V1 group (67.6, 49.5 and 17.6%, respectively) than those of control (97.3, 85.4 and 42.1%, respectively) and V2 (94.1, 78.8 and 40.9%, respectively) groups. In addition, the mean percentages of morphologically intact follicles in the V1 group were statistically lower than those in other groups, after transplantation. The rate of apoptosis in preantral follicles of the V1 group was significantly higher than that in the other groups. It was concluded that exposure of mice ovaries to SMF during vitrification resulted in greater resistance to injuries.


Main Subjects

  1. Posillico S, Kader A, Falcone T, et al. Ovarian tissue vitrification: Modalities, challenges and potentials. Curr Womens Health Rev 2010; 6(4):352-366.
  2. Shaw J, Jones G. Terminology associated with vitrification and other cryopreservation procedures for oocytes and embryos. Hum Reprod Update 2003; 9(6):583-605.
  3. Kojima SI, Kaku M, Kawata T, et al. Cranial suture-like gap and bone regeneration after transplantation of cryopreserved MSCs by use of a programmed freezer with magnetic field in rats. Cryobiology 2015; 70(3):262-268.
  4. Pang XF, Deng B. Infrared absorption spectra of pure and magnetized water at elevated temperatures. Europhys Lett 2011; 92(6):65001.
  5. Lee SYS, Sun CHB, Kuo TF, et al. Determination of cryoprotectant for magnetic cryopreservation of dental pulp tissue. Tissue Eng Part C Methods 2012; 18: 397-407.
  6. Ideta A, Hayama K, Urakawa M, et al. Cryopreservation of bovine biopsed embryo under a magnetic field. Reprod Fertil Dev 2006; 19(1): 178.
  7. Otero L, Rodríguez AC, Pérez‐Mateos M, et al. Effects of magnetic fields on freezing: Application to biological products. Compr Rev Food Sci Food Safe 2016; 15(3):646-667.
  8. Ghodbane S, Lahbib A, Sakly M, et al. Bioeffects of static magnetic fields: Oxidative stress, genotoxic effects, and cancer studies. Biomed Res Int 2013; 602987. doi: 10.1155/2013/602987.
  9. Rosen AD. Mechanism of action of moderate-intensity static magnetic fields on biological systems. Cell Biochem Biophys 2003;39(2):163-173.
  10. Behbahanian A, Eimani H, Zeinali B, et al. In vitro maturation, fertilization and embryo culture of oocytes obtained from vitrified auto-transplanted mouse ovary. Int J Fertil Steril 2013, (6) 278-285.
  11. Fathi R, Valojerdi MR, Eimani H, et al. Sheep ovarian tissue vitrification by two different dehydration protocols and needle immersing methods. Cryo letters 2011; 32(1):51-56.
  12. Liu J, Van der Elst J, Van den Broecke R, et al. Early massive follicle loss and apoptosis in heterotopically grafted newborn mouse ovaries. Hum Reprod 2002; 17(3):605-611.
  13. Kawata T, Abedini S, Kaku M, et al. Effects of DMSO (dimethyl sulfoxide) free cryopreservation with program freezing using a magnetic field on periodontal ligament cells and dental pulp tissues. Biomed Res 2012;23(3):438-443.
  14. Kyono K, Doshida M, Toya M, et al. New freezing method by pulsed magnetic field effects; whole ovaries of cynomolgus monkeys and rabbits. Reprod Biomed Online 2010; 20 (Suppl. 3): S12.
  15. Moriguchi H, Zhang Y, Mihara M, et al. Successful cryopreservation of human ovarian cortex tissues using supercooling. Sci Rep 2012; 2: 537.
  16. Stange BC, Rowland RE, Rapley BI, et al. ELF Magnetic field increase amino acid uptake into Vicia faba L. roots and alter ion movement across the plasma membrane. Bioelectromagnetics 2002; 33: 347-354.
  17. Antov Y, Barbul A, Mantsur H, et al. Electroendocytosis exposure of cells to pulsed low electric fields enhances adsorption and uptake of macromolecules. Biophys J 2005; 88: 2206-2223.
  18. Teodori L, Grabarek J, Smolewski P, et al. Exposure of cells to static magnetic field accelerates loss of integrity of plasma membrane during apoptosis. Cytometry 2002; 49:113-118.
  19. Rosen AD. A proposed mechanism for the action of strong static magnetic fields on biomembranes. Int J Neurosci 1993; 73(1-2):115-119.
  20. Rosen AD. Membrane response to static magnetic fields: Effect of exposure duration. Biochim Biophys Acta1993; 1148(2):317-320.