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

Mahsa Kheirollahi Kouhestani; Sayed Jamal Moshtaghian; Shahnaz Rezavi; MohammadHossein Nasr-Esfahani; MohammadReza Deemeh

Journal of Kermanshah University of Medical Sciences

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

Author(s):

Mahsa Kheirollahi Kouhestani¹,

Sayed Jamal Moshtaghian²,

Shahnaz Rezavi³,

MohammadHossein Nasr-Esfahani ^4,*,

MohammadReza Deemeh ⁵

1Dept. Physiology, School of Sciences, the University of Isfahan, Isfahan, Iran

2Dept. of Biology, the University of Isfahan, Isfahan, Iran

3Dept. of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4Dept. of Andrology and Embryology, Reproductive Medicine Research Center, Royan Institute, ACECR, Tehran, Iran

5Isfahan Fertility and Infertility Center, Isfahan, Iran

Journal of Kermanshah University of Medical Sciences:Vol. 13, issue 1; e79826

Published online:Jun 19, 2009

Article type:Research Article

Received:Oct 13, 2008

Accepted:Jun 02, 2009

How to Cite:Kheirollahi Kouhestani M, Moshtaghian S J, Rezavi S, Nasr-Esfahani M, Deemeh M. Evaluating the effectiveness of combination of Pure and Zeta methods in isolating mature sperms with normal chromatin structure.J Kermanshah Univ Med Sci.2009;13(1):e79826.

Abstract

Background: 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).

Methods: 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.

Results: 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 not differ in isolating sperms with regard to normal morphology, protamine content and DNA fragmentation (p>0.05).

Conclusion: 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..

Keywords

Zeta Method

Density Gradient Centrifugation

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