Showing 7 results for Viability
Somaye Khosravi Farsani, Reza Mahmoudi, Mir Abbas Abdolvahhabi, Mehdi Abbasi, Fateme Malek, Aligholi Sobhani,
Volume 5, Issue 5 (7-2007)
Abstract
Background: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimizing.
Objective: The aim of this study was to improve the single step and step-wise vitrification effects on maturing mouse GVBD oocytes by ethylene glycol (EG) in conventional straws.
Materials and Methods: Oocytes with compact cumulus cells were cultured for 3hr in TCM199 supplemented with 10% fetal bovine serum (FBS) in 5% CO2 in air. GVBD oocytes were randomly allocated into three groups. (1) Control (non-vitrified group), (2) exposed to single-step vitrification (contained of EG 20%+0.5M sucrose), (3) exposed to step-wise vitrification (2%, 5%, 10%, 20%EG +0.5M sucrose). In vitrification groups,oocytes were thawed and underwent additional 21 hr maturation. Viability of oocytes and maturation to MII stage were analyzed using inverted microscope and additionally by staining of propidium iodide and Hoechst 33342.
Results: All non-vitrified oocytes were viable after 24 hr; however, viability of vitrified samples in single-step group was significantly lower than that of the step-wise and control Groups. Also, the maturation rate in the step-wise group was significantly higher (p < 0.05) compared to single-step.
Conclusion: These results suggest that step-wise vitrification of GVBD oocytes as compared to single step vitrification was better in the rate of survival and in vitro maturation of oocytes.
Abolfazl Barkhordari, Seyedhossein Hekmatimoghaddam, Ali Jebali, Mohammad Ali Khalili, Alireza Talebi, Marzieh Noorani,
Volume 11, Issue 9 (12-2013)
Abstract
Background: The extensive use of different nanoparticles has raised great concerns about their occupational and biological safety.
Objective: The aim of this study was to evaluate the cytotoxic effect of zinc oxide nanoparticles (ZnO NPs) on viability of spermatozoa.
Materials and Methods: Semen samples were obtained from 15 healthy persons, and were analyzed using WHO guidelines. Each semen sample was separately incubated with different concentrations of ZnO NPs (10, 100, 500, and 1000 μg/mL) at 37PoPC for 45, 90, and 180 minutes. Then, the cell death percentage of spermatozoa was measured by MTT assay. Mann-Whitney test was used for comparison of different times and concentrations.
Results: The maximum cell death percentage was 20.8%, 21.2%, and 33.2% after 45, 90, and 180 minutes, respectively. In case of concentration, the highest concentration (1000 μg/mL) of ZnO NPs led to the highest toxicity for all incubation times. Statistically, there were significant differences in cell viability after 180 minutes vs. 45 and 90 minutes.
Conclusion: This study indicated that cytotoxicity of ZnO NPs is dose and time dependent.
Mohamed Sheata Ali Mohamed,
Volume 13, Issue 10 (10-2015)
Abstract
Background: Spermatozoa cryopreservation is used for the management of infertility and some other medical conditions. The routinely applied cryopreservation technique depends on permeating cryoprotectants, whose toxic effects have raised the attention towards permeating cryoprotectants-free vitrification technique.
Objective: To compare between the application of slow cryopreservation and vitrification on human spermatozoa.
Materials and Methods: This was an experimental controlled study involving 33 human semen samples, where each sample was divided into three equal parts; fresh control, conventional slow freezing, and permeating cryoprotectants-free vitrification. Viability and mitochondrial membrane potential (MMP) of control and post-thawing spermatozoa were assessed with the sperm viability kit and the JC-1 kit, respectively, using fluorescence-activated cell sorting analysis.
Results: Significant reduction of the progressive motility, viability and MMP was observed by the procedure of freezing and thawing, while there was not any significant difference between both cryopreservation techniques. Cryopreservation resulted in 48% reduction of the percentage of viable spermatozoa and 54.5% rise in the percentage of dead spermatozoa. In addition, high MMP was reduced by 24% and low MMP was increased by 34.75% in response to freezing and thawing. Progressive motility of spermatozoa was correlated significantly positive with high MMP and significantly negative with low MMP in control as well as post-thawing specimens (r=0.8881/ -0.8412, 0.7461/ -0.7510 and 0.7603/ -0.7839 for control, slow and vitrification respectively, p=0.0001).
Conclusion: Although both cryopreservation techniques have similar results, vitrification is faster, easier and associated with less toxicity and costs. Thus, vitrification is recommended for the clinical application.
Seyyed Mohammad Reza Hashemnia, Somayeh Atari-Hajipirloo, Shiva Roshan- Milani, Nasim Valizadeh, Sonya Mahabadi, Fatemeh Kheradmand,
Volume 14, Issue 9 (9-2016)
Abstract
Background: The anticancer agent imatinib (IM) is a small molecular analog ofATP that inhibits tyrosine kinase activity of platelet derived growth factors (PDGFs)and stem cell factor (SCF) receptor in cancer cells. However these factors have a keyrole in regulating growth and development of normal Sertoli, Leydig and germcells.
Objective: The aim of this study was to determine cell viability, PDGF and SCFlevels in mouse normal Sertoli cells exposed to IM.
Materials and Methods: In this experimental study, the mouse TM4 Sertoli cellswere treated with 0, 2.5, 5, 10 and 20 μM IM for 2, 4 or 6 days. The cell viabilityand growth factors levels were assessed by MTT and ELISA methods, respectively.For statistical analysis, One-Way ANOVA was performed.
Results: IM showed significant decrease in Sertoli cell viability compared to controlgroup (p=0.001). However, IM increased PDGF and SCF level insignificantly(p>0.05).
Conclusion: Results suggested that IM treatment induced a dose dependentreduction of cell viability in Sertoli cells. It seems that treatment with this anticancerdrug is involved in the fertility process. Further studies are needed to evaluate therole of PDGF and SCF in this cell.
Shirin Barati, Mansoureh Movahedin, Hossien Batooli,
Volume 16, Issue 5 (5-2018)
Abstract
Background: Spermatogonial stem cells are the foundation of spermatogenesis and male fertility. So, their maintenance and culture are very important.
Objective: In this study, we assessed protective effects of the Calligonum on in vitro viability and apoptotic and antiapoptotic genes expression of spermatogonial stem cells.
Materials and Methods: After 24 hr of culture, the spermatogonial stem cells were treated with 30 μM dose of H2O2 and then 10 μg/ml the Calligonum extract was added for 3 wks. Viability was assessed by Trypan blue, apoptosis using PI-Annexin and finally Bax, Bcl-2 and P53 genes expression by Real-Time Polymerase chain reaction.
Results: After 3 wk of treatment, viability in the Calligonum extract+H2O2 group was significantly higher than H2O2 group alone (p=0.001). In the Calligonum extract+H2O2 group, apoptosis, as well as expression of apoptotic genes (Bax and P53), was significantly lower than the group treated with H2O2 alone.
Conclusion: The results of this study showed that 30 μM H2O2 increased apoptosis but decreased viability in spermatogonial stem cells. Calligonum has antioxidant properties that can reduce apoptosis, Bax and P53 expression and increase the viability and Bcl-2 expression.
Hamid Reza Momeni, Mohammad Hussein Abnosi, Najmeh Eskandari,
Volume 18, Issue 11 (11-2020)
Abstract
Background: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay which evaluates cellular mitochondrial activity is widely used for the assessment of cell proliferation and viability.
Objective: This study was performed to assess human sperm viability using MTT assay.
Materials and Methods: In this laboratory study, human-ejaculated semen samples (n = 56 from different donors) were used. The sperm viability was determined using quantitative MTT assay and the sperm motility was assessed according to World Health Organization guidelines. Sperm viability and the correlation between sperm viability and motility were analyzed.
Results: Data revealed a marked positive correlation between MTT reduction rate and the percentage of viable spermatozoa. The Pearson's correlation coefficients also showed a significant correlation between sperm viability and motility.
Conclusion: MTT assay which is based on mitochondrial functionality is a reliable method for evaluating human sperm viability and could be used as a diagnostic test for predicting sperm fertilization ability in clinical settings.
Nabi A, Entezari F, Miresmaeili Sm, Lorian K, Anbari F, Motamedzadeh L,
Volume 19, Issue 5 (5-2021)
Abstract
Background: Polyvinylpyrrolidone (PVP) is a chemical material used in intracytoplasmic sperm injection program.
Objective: The aim of this study was to investigate the ideal time that sperm can be safely incubated in PVP with less structure and DNA damage.
Materials and Methods: Thirty-one oligoasthenoteratospermia samples were used. Sperm samples were prepared by discontinuous density-gradients method and incubated in 10% PVP at different time intervals (0, 5, 10, 15, 20, and 30 min). The effect of PVP was assessed on sperm DNA fragmentation and viability via sperm chromatin dispersion assay and Eosin-nigrosin staining.
Results: Data showed there was a significant increase in sperm DNA fragmentation after 10 min (36.76 ± 7.99, p < 0.001), 15 min (37.81 ± 8.11, p < 0.0001), 20 min (38.62 ± 8.00, p < 0.0001), and 30 min (40.05 ± 7.69, p < 0.0001) compared to 0 min. The viability rate also significantly reduced after 10 min (57.71 ± 10.85, p = 0.04), 15 min (55.81 ± 10.87, p < 0.0001), 20 min (53.19 ± 11.44, p < 0.0001), and 30 min (50.24 ± 11.81, p < 0.0001) compared to 0 min.
Conclusion: As a result, sperm samples could be incubated with PVP for 10 min with less DNA damage. While, prolonged incubation may significantly damage the sperm DNA integrity and viability.
