[Evaluating the effectiveness of combination of Pure and Zeta methods in isolating mature sperms with normal chromatin structure]

ABSTRACT

Isolating sperms with normal chromatin content is considered vital in Intra Cytoplasmic Sperm Injection treatment. Today, researchers are trying to find new procedures to select sperms with normal chromatin content and morphology. This study examines the effectiveness of Zeta, Density Gradient Centrifugation [Pure Sperm] and combination of Pure-Zeta procedures in selecting sperms with normal chromatin structure using Sperm Chromatin Dispersion [SCD], and Chromomycin A3 [CMA3]. 28 patients participated in the study. Semen samples were analyzed according to WHO criteria. The samples were then divided into four equal portions of control, Zeta method, DGC [Pure Sperm] method and Pure-Zeta method. Sperm morphology [Diff Quick Staining], protamine deficiency [CMA3] and DNA integrity [SCD] were carried out for all the samples. The results were analyzed using ANOVA. P< 0.05 was considered significant. There was a significant reduction in the percentage of sperm protamine deficiency and DNA fragmentation in Zeta, Pure and Pure-Zeta method groups compared to the control group [p <0.05]. In terms of a normal morphology, Pure and Pure-Zeta procedures proved significantly different from the control group [p<0.05 normal morphology]. Zeta, Pure and Pure-Zeta procedures did no differ in isolating sperms with regard to normal morphology, protamine content and DNA fragmentation [p > 0.05]. Based on the result, Zeta and Pure methods improved selection of sperms with normal morphology, protamine content and DNA integrity. Yet, the combination of Pure-Zeta was more effective in terms of isolating spermatozoa with normal morphology, protamine content and DNA integrity than when Zeta and Pure methods were implemented individually