Abstract
Limited information exists about the effects of antioxidants on sperm motility and DNA integrity during vitrification in humans. This study compared the effects of reduced glutathione (GSH) and monothioglycerol (MTG) at different concentrations on post-thaw sperm motility and DNA integrity after vitrification in humans using 0.25 M trehalose as a cryoprotective agent, and found that supplementation of MTG or GSH at 0.5 mM resulted in significantly higher (P < 0.05) recovery rates of post-thaw total and progressive motilities. GSH was more powerful than MTG at the same concentration in cryoprotecting sperm motility (38.9% ± 3.6% vs 32.8% ± 2.4% compared to the control 26.8% ± 2.1% in recovery rate of progressive motility), but both had no significant influence on sperm DNA integrity during vitrification. It was concluded that sperm motility is more sensitive to oxidative stress during vitrification than sperm nuclear DNA, and supplementation of MTG or GSH in vitrification medium is beneficial in cryoprotecting sperm motility.
Highlights
Sperm cryopreservation has long been an important strategy in fertility preservation and assisted reproduction
This study compared the effects of reduced glutathione (GSH) and monothioglycerol (MTG) at different concentrations on post-thaw sperm motility and DNA integrity after vitrification in humans using 0.25 M trehalose as a cryoprotective agent, and found that supplementation of MTG or GSH at 0.5 mM resulted in significantly higher (P < 0.05) recovery rates of post-thaw total and progressive motilities
We measured DNA fragmentation index (DFI) of post-thawed sperm and the data summarized in Figure 4 indicates that MTG had no significant effect (P > 0.05) on sperm DNA integrity during vitrification at the concentrations tested compared to the control DFI (13.5% ± 1.8%)
Summary
Sperm cryopreservation has long been an important strategy in fertility preservation and assisted reproduction. Studies have shown that antioxidants quercetin, Vitamin E, melatonin, reduced glutathione (GSH) and MitoTEMPO are beneficial to improve post-thaw sperm motility after slow freezing [5] [6] [7] [8] and rapid freezing [9] [10] [11] [12]. Quercetin and GSH were reported to be efficient in protecting the integrity of sperm nuclear DNA during slow freezing [6] [7] [8] [10] and rapid freezing [9] [11]. Only butylhydroxytoluene (BHT, a synthetic analogue of vitamin E) and hypotaurine were reported to be beneficial in vitrifying human sperm motility and DNA integrity [16] [17]. Antioxidant monothioglycerol (MTG) has been found to be extremely effective in cryoprotecting mouse sperm motility, DNA integrity and fertilization ability [18] [19] [20]; this antioxidant has not yet been tested for human sperm cryopreservation
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