P-97: The Effects of Catalase Addition to The Cryopreservation Medium on Follicles Apoptosis and Oxidative Stress in Human
Today, cryopreservation of ovarian tissue has been the effective procedure to restore fertility in can cer patients. It is well known that oxidative stress (OS) is a widespread phenomenon that occurs in ovary cryopreservation. Numerous studies show that follicular atresia in mammalian species due to the accumulation of toxic metabolites often results from oxidative stress. Therefore, currently there is a great interest to the use of antioxidants to prevent ROS generation and ROS-induced apoptosis during the ovary cryopreservation processes. It has been revealed that including catalase as an antioxidant in sperm freezing medium reduced lipid peroxidation. In this regards, the aim of this study was to investigate the effects of catalase addition to the cryopreservation medium on cell morphology, viability, apoptosis, ROS generation and lipid peroxidation in human ovary.
Materials and methods
Biopsies of ovarian cortex from cancer patient (n=14) were divided into 4 groups: without catalase, with catalase in freezing medium, with catalase in thawing medium and with catalase in both medium. After 2 weeks the morphology, viability and incidence of apoptosis were evaluated using Hematoxylin and eosin (H & E), Calcein- AM and Ethidium homodimer-1staining and TUNEL. H2O2 generation and LPO were assessed by means of DCFHDA fluorescence and MDA assay kit.
Compared with the control group, cryopreservation with catalase resulted in significant decrease in the percentage of DCFH-DA fluorescence, apoptosis, and increase in viability of the follicles. There was no difference between groups in follicular morphology.
Catalase addition to cryopreservation medium can be a potential tool against oxidative damage and apoptosis in human ovarian tissues.