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Mohamad Reza Darabi, Mohamad Hosein Nasr-Esfahani, Hosein Baharvand, Mohmad Mardani, Hojatolah Karimi-Jashni,
Volume 6, Issue 5 (7-2008)

Background: The values of embryonic stem cell and cloning are evident. Production of clone from embryonic stem cells can be achieved by introduction of stem cell into a tetraploid blastocyst. Tetraploid blastocyst can be produced in vitro by electrofusion of 2-cell embryos.
Objective: The aim of this study was to assess the effect of different voltages and durations on fusion rate of bovine 2-cell embryos and their subsequent development in vitro.   
Material and Methods: The in vitro produced bovine 2-cell embryos were categorized into 3 groups: (1) fused group (FG); 2-cell embryos fused by exposure to different voltages (0.5, 0.75, 1, 1.25 and 1.5 kV/cm) and durations (20, 40, 60, 80 and 100 μs), (2) exposed control group (ECG);  2-cell embryos exposed to different voltages and durations but remained unfused and (3) unexposed control group (UCG); embryos cultured without exposure to any voltage. The embryos from each group were cultured and fusion, cleavage and developmental rates were compared in each group.
Results: The results show that increased voltage, increases the fusion rate up to 88% for 1.5 kV/cm; however, the rate of cleavage and blastocyst formation decreases significantly to 18% and 10% respectively (p<0.05). Increased duration does not significantly increase fusion rate, however, in high voltage, increased duration decreases cleavage rate and blastocyst formation rate. Blastocyst formation rate in UCG showed a better development (32%) compared to FG (20%) or ECG (22.5%) (p<0.05).
Conclusion: It can be concluded that for optimal fusion, cleavage and development, one pulse of 0.75 kV/cm for 60μs should be applied.
Noush Afarin Khajavi, Shahnaz Razavi, Mohammad Mardani, Marziyeh Tavalaee, Mohammad Reza Deemeh, Mohammad Hossein Nasr-Esfahani,
Volume 7, Issue 2 (7-2009)

Background: Sperm selection for ICSI based on morphology and motility might not be relevant to chromatin integrity. Thus sperm selection based on sperm characteristics has been suggested.
Objective: The aim of this study was to compare the efficiency of Zeta method with routine Density Gradient Centrifugation method (DGC) for the selection of sperm with higher DNA integrity.
Materials and Methods: Semen samples were obtained from 63 individuals referring to Andrology Unit of Isfahan Fertility and Infertility Center. Semen analysis was carried out according to WHO criteria. Each semen sample was divided into three equal portions. One portion was used as control, the second portion was used for Zeta method and the third portion underwent DGC method. Each portion was evaluated to DNA integrity by TUNEL assay. Student t-test was carried out using SPSS and p-value lower than 0.05 was considered significant.
Results: The mean number of sperm DNA fragmentation in Zeta and DGC methods were significantly decreased compare to the control group (p<0.001). In addition, Zeta method was more efficient than the DGC method in the selection of sperm with intact DNA (p<0.001).
Conclusion: The Zeta method appears to be a suitable procedure to recover sperm with normal DNA integrity.
Mohammad Mardani, Ahmad Vaez, Shahnaz Razavi,
Volume 12, Issue 5 (6-2014)

Background: Currently, relation between reactive oxygen species (ROS) ROS concentration and semen quality was indicated. Saffron has traditionally been not only considered as a food additive but also as a medicinal herb, which has a good antioxidant properties.
Objective: The aim of this study was to evaluate the protection potency of saffron and vitamin E on sperm chromatin integrity.
Materials and Methods: Thirty adult male Wistar rats divided equally into saffron (100 mg/kg), vitamin E (100 mg/kg) and control (0.5cc distilled water /day) groups. After 60 days, cauda epididymis dissected and sperm cells were used for analysis of sperm chromatin packaging by chromomycin A3 (CMA3) staining, and sperm chromatin susceptibility to acid denaturation by acridine orange (AO) staining.
Results: The mean percentage of CMA3 positive sperm was significantly decreased in saffron and vitamin E groups relative to control group (p<0.001). Moreover, the AO staining results showed that the mean percentage of sperm with DNA damage was significantly decreased in saffron and vitamin E groups as compared with control group (p<0.001).
Conclusion: Our results purposed that saffron can protect sperm against DNA damage and chromatin anomalies.
Farahnaz Mardanian, Moones Kazeroonizadeh, Bahman Rashidi,
Volume 13, Issue 9 (10-2015)

Background: Infertility is an increasing medical and social problem. In vitro fertilization (IVF) has become a common and accessible treatment for a wide variety of indications that have variable outcomes. Natural killer (NK) cells have been identified as relevant immunological factors involved in reproductive success or failure.
Objective: The aim of this study was to compare the percentage of peripheral blood CD56+ (CD56dim and CD56bright) cells and the level of NK cell in patients with IVF failure with those of successful IVF control women.
Materials and Methods: We assessed the level of CD56dim CD16+ and CD56bright CD16- cells in 50 women under IVF treatment and compared between successful IVF and IVF failure with the flowcytometry technique.
Results: Of studied women, 68% did not response to IVF therapy and 32% had successful IVF, the level of CD56dim CD16+ cells in women with IVF failure was significantly higher than successful IVF (p<0.0001) but the level of CD56bright CD16- cells was not significantly different between women with IVF failure and successful IVF (p=0.28).
Conclusion: The results of present study demonstrated that the level of NK cells as a risk factor is associated with pregnancy loss in women with IVF failure. However, number of sample in this study is low and further studies with more sample size are needed to be done. We suggest considering treatment option for women undergoing repeated IVF failure with increased percentage of CD56dim cells and the level of peripheral blood NK cell.

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