Evaluation of Cryopreservation of Bovine Ovarian Tissue by Analysis of Reactive Species of Oxygen, Toxicity, Morphometry, and Morphology

Abstract

Ovarian tissue cryopreservation has been widely investigated for preserving female fertility. In the present study, we aimed to compare the effects of three concentrations (1, 1.5, and 3 M) of dimethylsulfoxide (DMSO) on the vitrification of ovarian tissue. The ovarian cortex was divided into control and vitrified groups: (i) 1 M-DMSO, (ii) 1.5 M-DMSO, and (iii) 3 M-DMSO. Follicles from all fragments were analyzed for DMSO-induced deleterious effects, morphological and morphometric aspects, and concentration of reactive oxygen species. Additionally, the fragments were cultured to assess the integrity and return of follicular development post-vitrification. All DMSO concentrations resulted in a higher percentage of degenerated preantral follicles than before the cryopreservation process. After vitrification, the cryopreserved ovarian fragments showed similar percentages of intact follicles; however, the 3 M DMSO concentration differed from the control. Analyzing free radical production, we found that the 3 M DMSO concentration had higher levels of oxidative stress than the lower DMSO. After in vitro cultivation of the vitrified/warmed fragments, the 1 M DMSO concentration exhibited higher percentages of morphologically intact follicles than the other concentrations. Therefore, we suggest that bovine preantral follicles can be cryopreserved in situ with greater efficiency in 1 M DMSO.

Keywords

• vitrification
• DMSO
• in vitro culture of preantral follicles
• ROS