Exogenous Melatonin Ameliorates the Negative Effect of Osmotic Stress in Human and Bovine Ovarian Stromal Cells

Abstract

Ovarian tissue cryopreservation transplantation (OTCT) is the most flexible option to preserve fertility in women and children with cancer. However, OTCT is associated with follicle loss and an accompanying short lifespan of the grafts. Cryopreservation-induced damage could be due to cryoprotective agent (CPA) toxicity and osmotic shock. Therefore, one way to avoid this damage is to maintain the cell volume within osmotic tolerance limits (OTLs). Here, we aimed to determine, for the first time, the OTLs of ovarian stromal cells (OSCs) and their relationship with reactive oxygen species (ROS) and mitochondrial respiratory chain activity (MRCA) of OSCs. We evaluated the effect of an optimal dose of melatonin on OTLs, viability, MRCA, ROS and total antioxidant capacity (TAC) of both human and bovine OSCs in plated and suspended cells. The OTLs of OSCs were between 200 and 375 mOsm/kg in bovine and between 150 and 500 mOsm/kg in human. Melatonin expands OTLs of OSCs. Furthermore, melatonin significantly reduced ROS and improved TAC, MRCA and viability. Due to the narrow osmotic window of OSCs, it is important to optimize the current protocols of OTCT to maintain enough alive stromal cells, which are necessary for follicle development and graft longevity. The addition of melatonin is a promising strategy for improved cryopreservation media.

Melatonin activates Nrf2/HO‐1 signalling pathway to antagonizes oxidative stress‐induced injury via melatonin receptor 1 (MT1) in cryopreserved mice ovarian tissue

“…Once the oxidases and antioxidant enzymes are out of balance, a large amount of ROS will be produced, causing oxidative stress and damaging tissues [8]. According to research, on bovine ovarian stromal cells, MLT may lessen the adverse consequences of osmotic pressure by increasing the protein expression of CAT and SOD (Asadi et al., 2022). This aligns with the findings of our research.…”

Rational synthesis of total damage during cryoprotectant equilibration: modelling and experimental validation of osmomechanical, temperature, and cytotoxic damage in sea urchin (Paracentrotus lividus) oocytes

“…Indeed, Wang et al (2011) found accumulated osmotic damage was temperature specific while Williams & Takahashi (1982) found membrane lysis of sea urchin eggs was mitigated at lower temperatures during hypertonic challenge. Supplements that prevent membrane and cytoskeletal regulation may reduce time-dependent osmotic damage ( Asadi, Najafi & Benson, 2022 ). However, there may be a trade-off between time-dependent osmotic damage and sensitivity to lysis during osmotic challenge, or loss of cell functionality with supplements that depolarizes F-actin, such as cytochalasin-D or cytochalasin-B ( Fujihira, Kishida & Fukui, 2004 ; Takamatsu et al, 2005 ; Wang et al, 2016 ).…”

3D hUC-MSC spheroids exhibit superior resistance to autophagy and apoptosis of granulosa cells in POF rat model

“…Therefore, POF has a significant effect on the reproductive health and quality of life of women. Current research suggests that the primary cause of POF is low ovarian reserve function due to atresia of the dominant follicles and failure of the primordial follicles in the ovary due to oxidative stress (Sun et al 2018, Asadi et al 2022, Jiang et al 2019. GCs play a crucial role in follicular development and maturation by producing estradiol and other maturation-promoting factors (Zhu et al 2022).…”