Vitrification technique for female germinative tissue cryopreservation and banking

Area of Science:

• Reproductive biology
• Cryobiology
• Tissue engineering

Background:

• Ovarian tissue cryopreservation is crucial for fertility preservation in women facing sterilizing medical treatments.
• Vitrification offers a promising method for ovarian tissue banking, but requires optimized protocols and devices.
• Assessing the morphological and quantitative integrity of ovarian follicles post-vitrification is essential.

Purpose of the Study:

• To evaluate a novel device and protocol for the vitrification and warming of human ovarian germinative tissue.
• To assess the morphological integrity and follicular count of vitrified-ovarian tissue compared to fresh tissue.

Main Methods:

• Human ovarian cortical tissue fragments were obtained.
• Tissue fragments underwent either histological analysis (control) or vitrification/warming using a specialized device and protocol.
• Morphological analysis (microscopy) and follicular counting (primordial and primary) were performed on fresh and vitrified/warmed tissues.
• Statistical comparison using the Mann-Whitney test (p<0.05) was conducted.

Main Results:

• Ovarian follicles in various maturation stages were observed in both fresh and vitrified/warmed tissues, with a majority being healthy primordial and primary follicles.
• Vitrified/warmed tissue showed some stromal edema and hydropic degeneration but no necrosis or autolysis.
• No statistically significant difference in the number of primordial and primary follicles was found between fresh and vitrified/warmed tissue fragments (p>0.05).

Conclusions:

• The developed vitrification device and protocol effectively preserved human ovarian germinative tissue.
• Significant morphological changes were not observed between fresh and vitrified/warmed tissues.
• This method is a viable option for ovarian tissue banking to restore fertility in women undergoing sterilizing treatments.