O-7: The Comparison of Efficiency of Density Gradient Centrifugation and Zeta Methods in Separation of Mature and Normal Spermatozoa by TUNEL, SCD, Acridine Orange and CMA3 (Pages: 0-0)


Nasr Esfahani MH , Deemeh M , Tavalaee M , Razavi Sh , Khajavi N* *,

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Objective: The aim of this study was to examine the efficiency of Zeta method and Density Gradient Centrifugation method for the selection of normal sperms by TUNEL, Sperm Chromatin Dispersion (SCD), Acridine Orange (AO) and Chromomycin A3 (CMA3). Materials and Methods: The study was conducted on 63 patients. Semen analysis was carried out according to WHO criteria. Semen samples were divided into three equal portions. One portion was left untreated (control), the second portion was used for Zeta method and the third portion underwent DGC method. Then, all of the portions were evaluated to sperm morphology (Diff Quick staining ), protamine deficiency (CMA3) and DNA integrity ( SCD, AO and TUNEL). Coeficients of correlation and student t-test were carried out using SPSS and P-value lower than 0.05 was considered significant. Results: The mean number of sperm abnormality, protamine deficiency and DNA fragmentation in Zeta and DGC methods were significantly decreased as compared to the control group (p<0.005). In addition, Density Gradient Centrifugation method was superior to Zeta method in the selection of sperms with normal morphology (p<0.005) and Zeta method was superior to DGC method in the selection of sperms with intact DNA (p<0.005). Conclusion: Both Zeta and DGC methods were effective in the selection of sperm with better quality in terms of normal morphology, normal protamine content and DNA integrity. However, Density Gradient Centrifugation method was superior to Zeta method in the selection of sperms with normal morphology and Zeta method was superior to DGC method in the selection of sperms with intact DNA.