A Mohseni Meybodi1, H Mozdarani, Sh Zari Moradi, M R Akhoond. 1. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. anahitamohseni@gmail.com
Abstract
PURPOSE: The aim of this study was to investigate whether treatment of sperm from infertile patients would gluthatione could reduce sperm premature chromosome condensation (PCC). To reach the goals of this study, the frequency of sperm PCC formation in sperm of normal and sub-fertile men with/without glutathione treatment were compared and analyzed. METHODS: Hamster oocytes were retrieved after super ovulation by PMSG and HCG injection. Following treatment with hyaluronidase, zonae was removed by trypsin digestion. Sperm were classified into 3 groups according to morphology, movement and counts, treated with glutathione(10 μg/ml) and then processed by swim up method. After capacitation, zona-free oocytes were incubated with sperm then transferred to fresh media containing colcemid. Cells were fixed and slides prepared using Tarkowskie's standard air drying technique and after staining in 5% Giemsa, oocytes were analyzed at high magnification. RESULTS: Sperm penetration rate was higher and the rate of intact sperm head and PCC was lower in GSH treated samples compared to non treated groups. Sperm penetration rate was significantly higher in treated astheno sperm samples compared to non-treated ones (66.4% and 50. 97% respectively) (P < 0.001). We observed a significantly higher PCC frequency in infertile patients (P < 0.001). In addition, there was a significantly lower rate of intact sperm head in treated astheno sperm samples (17.49%) compared to non treated ones (26.79%) (P < 0.001). Finally, a significantly lower rate of PCC in treated astheno sperm samples comparing to non treated ones was seen (51.06% and 72.96% respectively) (P < 0. 001). CONCLUSIONS: Our results show that sperm PCC formation could be one of the major causes of failed fertilization in individuals with sperm abnormalities. Also sperm PCC formation may be involved in the etiology of some cases of idiopathic infertility. Given that the susceptibility of sperm to oxidative stress is significantly greater in idiopathic infertile men, our results show that treatment with glutathione could significantly reduce these stress factors and increase ART outcome.
PURPOSE: The aim of this study was to investigate whether treatment of sperm from infertilepatients would gluthatione could reduce sperm premature chromosome condensation (PCC). To reach the goals of this study, the frequency of sperm PCC formation in sperm of normal and sub-fertile men with/without glutathione treatment were compared and analyzed. METHODS:Hamster oocytes were retrieved after super ovulation by PMSG and HCG injection. Following treatment with hyaluronidase, zonae was removed by trypsin digestion. Sperm were classified into 3 groups according to morphology, movement and counts, treated with glutathione(10 μg/ml) and then processed by swim up method. After capacitation, zona-free oocytes were incubated with sperm then transferred to fresh media containing colcemid. Cells were fixed and slides prepared using Tarkowskie's standard air drying technique and after staining in 5% Giemsa, oocytes were analyzed at high magnification. RESULTS: Sperm penetration rate was higher and the rate of intact sperm head and PCC was lower in GSH treated samples compared to non treated groups. Sperm penetration rate was significantly higher in treated astheno sperm samples compared to non-treated ones (66.4% and 50. 97% respectively) (P < 0.001). We observed a significantly higher PCC frequency in infertilepatients (P < 0.001). In addition, there was a significantly lower rate of intact sperm head in treated astheno sperm samples (17.49%) compared to non treated ones (26.79%) (P < 0.001). Finally, a significantly lower rate of PCC in treated astheno sperm samples comparing to non treated ones was seen (51.06% and 72.96% respectively) (P < 0. 001). CONCLUSIONS: Our results show that sperm PCC formation could be one of the major causes of failed fertilization in individuals with sperm abnormalities. Also sperm PCC formation may be involved in the etiology of some cases of idiopathic infertility. Given that the susceptibility of sperm to oxidative stress is significantly greater in idiopathic infertilemen, our results show that treatment with glutathione could significantly reduce these stress factors and increase ART outcome.