Reduced glutathione in the freezing extender improves the in vitro fertility of ram epididymal spermatozoa

Hajihassani, Aazam; Ahmadi, Ebrahim; Shirazi, Abolfazl; Shams‐Esfandabadi, Naser

doi:10.1016/j.smallrumres.2019.02.016

Cited by 6 publications

(4 citation statements)

References 38 publications

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“…These altered chromatin gametes are susceptible to the attack by free radicals which finally leads to reducing the number of spermatogonia, spermatocytes, spermatids and spermatozoa (Houston et al, 2018). Also, high‐levels of ROS affect Ca ++ channels, reduce glutathione levels and sperm adenosine triphosphate reserves, therefore, can decrease spermatozoa mobility (Hajihassani et al, 2019; Mannowetz et al, 2018). In the present study, dose and treatment period played an important role in the changes of testicular tissue and seminiferous tubules.…”

Section: Discussionmentioning

confidence: 99%

Neutralizing the adverse effects of cyclophosphamide on the mouse testis and sperm parameters through pentoxifylline: A molecular and stereological study

2022

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Cyclophosphamide (CP) is one of the chemotherapeutic drugs, which plays its role by interfering with all rapidly proliferating tissues like cancer and testis. The aim of this study was to investigate the protective effect of pentoxifylline (PTX) on the sperm parameters, spermatogenesis indices, biochemical alterations and gene expressions, in adult male mice treated with CP. A total of 24 male NMRI mice were divided into four groups: control, CP group (15 mg/kg weekly), PTX (100 mg/kg daily) and CP + PTX and treated for 35 days with the intraperitoneal injection. A significant decrease in the spermatogenesis indices, Leydig cells, sperm motility, viability, count, tail length and daily sperm production was found in the CP group compared to the control group. The results of this study indicate that PTX prevented these adverse effects of CP and decreased the number of apoptotic cells. Moreover, the CP group showed decreased levels of total antioxidant capacity, testosterone, lipid peroxidation and the expression of cytochrome P450 and 3β‐hydroxysteroid, all of which were neutralized in the CP + PTX group. It seems that PTX has the potential to be used in therapeutic regimens of cancer patients to reduce the side effects of CP. However, more research is needed to evaluate this prevention in mice models of cancer.

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Section: Discussionmentioning

confidence: 99%

Neutralizing the adverse effects of cyclophosphamide on the mouse testis and sperm parameters through pentoxifylline: A molecular and stereological study

2022

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“…The spermatozoa were prepared according to Hajihassani et al. ( 2019 ). Briefly, 200 μl of frozen‐thawed epididymal sperm suspension was placed on 1 ml Histoprep and centrifuged at 300 × g for 5 min.…”

Section: Methodsmentioning

confidence: 99%

“…After in vitro maturation, the oocytes were transferred to the droplets of IVF-TALP and incubated at 39 • C with 5% CO 2 in humidified atmosphere until sperm addition. The spermatozoa were prepared according to Hajihassani et al (2019). Briefly, 200 µl of frozen-thawed epididymal sperm suspension was placed on 1 ml Histoprep and centrifuged at 300 × g for 5 min.…”

Section: In Vitro Embryo Productionmentioning

confidence: 99%

The effect of chemical treatment of the sheep embryo zona pellucida on the ability of blastocysts to hatch after vitrification and warming

Mousavi

Ahmadi

Shirazi

et al. 2021

Veterinary Medicine & Sci

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Background:The embryo release from the zona pellucida is of prerequisites of successful implantation.Objectives: Regarding the negative impact of embryo cryopreservation on the blastocysts hatchability, the aim of the present study was to investigate the effects of treating embryonic zona pellucida with pronase or acidic Tyrode's solution (ATS) before morula formation on the viability, freezability, and hatchability of vitrified-warmed resulted blastocysts. Methods:In the first experiment, the zona pellucida of 3-and 4-day-old embryos were treated with the above compounds for 30 or 45 s. Then, the competency of the treated embryos to reach to blastocyst stage and the hatchability of resulting blastocysts were investigated. In the second experiment, the cryo-survivability and hatching rate of blastocysts resulting from 3-day-old embryos treated with pronase and ATS for 30 s were tested. Results:In the first experiment and in contrast to the 45 s exposure, 30-s exposure of embryos to pronase or ATS did not have negative effect on the viability and development of embryos to blastocyst stage. In the second experiment, the freezability of blastocysts derived from 3-day-old embryos treated with pronase and ATS for 30 s was not different from that of the control group. However, the hatching rate of the pronase group was significantly higher than that of the control group. Conclusion:The results of the present study showed that reducing the thickness of zona pellucida of sheep embryos with pronase had no negative effect on the developmental competency and freezability of the treated embryos and improved the hatchability of vitrified-warmed blastocysts.

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“…Two slides per each sample were prepared and 200 spermatozoa were counted on each slide. The percent of spermatozoa with intact membrane was calculated by dividing the number of spermatozoa with swollen tail (intact membrane) to the total counted sperm 27 …”

Section: Methodsmentioning

confidence: 99%

Ram epididymal sperm frozen in an extender containing ethylene glycol have higher post‐thaw longevity and in vitro fertility

et al. 2021

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Background: Establishing an efficient, simple and inexpensive method for freezing ram epididymal sperm so that the quality and fertility of spermatozoa could be maintained for a longer period after thawing is of great practical value. Objectives:To optimize freezing and thawing protocol for ram epididymal sperm using either ethylene glycol (EG) or glycerol (GLY) as cryoprotectants (CPAs). Then, to evaluate the post-thaw longevity and in vitro fertility of spermatozoa that were frozen and thawed according to the optimized protocol. Materials and methods:At first, an optimum protocol for freezing and thawing sperm using EG or GLY were investigated, and the next experiments were performed using the spermatozoa that had been frozen and thawed according to the optimized protocol for each CPA. In the next experiments, frozen-thawed and fresh sperm were diluted in an isotonic culture medium and subsequently incubated at 39 • C for 4 h. The motility characteristics and functional membrane integrity (FMI) of spermatozoa were evaluated after thawing, after dilution (t 0 ), and after incubation (t 4 ). The in vitro fertility of the spermatozoa was assessed at t 0 and t 4 .Results: For both CPAs, the highest motility parameters and FMI was found for spermatozoa frozen at 3 cm above LN2 and thawed at 50 and 65 • C (P < 0.05). In comparison to the spermatozoa of GLY group, the spermatozoa of the EG group had higher total and progressive motility at t 0 , as well as higher FMI, total and progressive motility, and linearity at t 4 (P < 0.05). Fertility of frozen-thawed sperm was higher than that of fresh sperm at t 0 (P < 0.05). Incubation treatment increased the fertility of fresh sperm while decreased the fertility of frozen-thawed sperm, and this decline was more severe in GLY than in the EG group. Conclusion:Based on the findings, EG can be a more suitable CPA for freezing ram epididymal sperm.

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Reduced glutathione in the freezing extender improves the in vitro fertility of ram epididymal spermatozoa

Cited by 6 publications

References 38 publications

Neutralizing the adverse effects of cyclophosphamide on the mouse testis and sperm parameters through pentoxifylline: A molecular and stereological study

Neutralizing the adverse effects of cyclophosphamide on the mouse testis and sperm parameters through pentoxifylline: A molecular and stereological study

The effect of chemical treatment of the sheep embryo zona pellucida on the ability of blastocysts to hatch after vitrification and warming

Ram epididymal sperm frozen in an extender containing ethylene glycol have higher post‐thaw longevity and in vitro fertility

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