The Comparison Of Efficiency Of Density Gradient Centrifugation And Zeta Methods In Separation Of Mature Sperm With Normal Chromatin Structure

Document Type : Original Article


1 Reproduction and Development Department, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran

2 Isfahan Fertility and Infertility Center, Isfahan, Iran

3 Anatomy Department, Isfahan University of Medical Sciences, Isfahan, Iran


Objective: To examine the efficiency of both the Zeta and density gradient centrifugation (DGC) methods for the selection of normal chromatin sperm by TUNEL, sperm chromatin dispersion (SCD), acridine orange and chromomycin A3 (CMA3).
Materials and Methods: The study was performed on 63 patients. Semen analysis was carried out according to WHO criteria. Semen samples were divided into three equal portions. One portion was used as the control, the second portion was used for the Zeta method and the third portion underwent DGC method. All portions were evaluated for sperm morphology (Diff Quick staining), protamine deficiency (CMA3) and DNA integrity (SCD, AO and TUNEL). Coefficients of correlation and students t test were carried out using SPSS 11.5.
Results: The mean percentage of abnormal sperm, protamine deficiency and DNA fragmentation in the Zeta and DGC methods were significantly decreased as compared to the control group (p<0.005). In addition, the DGC method was higher in comparison to the Zeta method in the selection of sperm with normal morphology (p<0.005). The Zeta method was higher in comparison to the DGC method in the selection of sperm 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. Therefore, we suggested that the combined Zeta and DGC method should be used for selection of normal sperm.