Comparison of survival and apoptosis parameters of ovarian tissue follicles in two vitrification methods of needle-immersion and cryo-support in cancer patients.

Abstract

Objective: Ovarian tissue (OT) cryopreservation is a useful technique for preserving fertility potential in women with cancer. Vitrification is a relatively new method. Several devices were discussed. Among these methods, cryo-support and needle-immersion vitrification (NIV) stand out as particularly popular. The aim of this study is the comparison of these devices in terms of follicle quality and DNA status.

Materials and methods: The OT from 20 cancer patients was transferred with Dulbecco's Phosphate-Buffered Saline supplemented with 5% serum and maintained at 4 °C for 1 hour. After preparation of OT vitrified by two freezing solutions with different concentrations of cryoprotectant, small fragments of OT (~5×1×1 mm) were attached to an insulin needle, and in another group, the tissue fragments (~5×5×1 mm) were loaded onto the Ova Cryo Device Type M, also called cryo-support, and placed in liquid nitrogen. After warming, small tissue pieces were prepared to assess the quality of primordial, primary, and secondary follicles using eosin-hematoxylin staining. An immune-histochemical investigation, including anti-p53 and anti-Caspase 3, was conducted to examine the apoptosis pathway.

Results: The data showed a larger number of high-quality follicles in the cryo-support group (p<0.05). Also, the level of apoptosis (p53) molecules is higher in the NIV method (p<0.05). However, the levels of caspase 3 were not significantly different between the NIV approach and the cryo-support vitrification group.

Conclusion: A recent study revealed that the cryo-support device is more effective in increasing high-quality follicles and reducing p53, which is associated with early stages of apoptosis. This device may improve clinical outcomes and may be recommended for cryopreservation programs.