Abstract
This was a prospective multicenter study aiming at comparing the efficiency of sperm selection by density gradient centrifugation (DGC) in reducing sperm DNA fragmentation (SDF) in different ART centers. The study was designed using 290 semen samples collected from 10 different ART centers performing artificial insemination, in vitro fertilization and blind assessment of SDF at the University facilities. The results showed that while there was a significant reduction in the SDF levels in sperm isolated from the gradient centrifuged pellet (DGC) compared to neat semen samples (NSS), there was also significant inter-center variability in the efficiency to reduce SDF values by DGC (78.5% to 29.2%). Surprisingly, for some patients, the level of SDF actually increased following sperm selection. The main conclusions derived from this study were that 1) isolation of sperm from the gradient pellet by DGC must be performed using validated, optimized protocols; 2) routine comparison of SDF values in NSS semen and in processed sperm after DGC or swim-up must be recommended as part of the internal quality control (QC) of ART laboratories to test the efficacy of sperm processing; and 3) SDF values in processed spermatozoa should be obtained to compare with the pregnancy rate when insemination or fertilization is about to be performed, otherwise, attempts to predict pregnancy outcome from SDF could be biased or are essentially meaningless.
Highlights
Predicting the success rate of human pregnancy using estimates of sperm DNA damage to date has been a very controversial issue, and that is no doubt further complicated by a plethora and permutation of techniques and procedures that vary in competency and delivery from laboratory to laboratory, all of which result in a fundamental lack of standardization
Given the wide range of variables that contribute to the success of an ART outcome, it is not surprising that this variability is reflected in the ability of sperm DNA fragmentation (SDF) to predict pregnancy outcome
In order to improve the predictive power of SDF it is important that we account and describe the precise conditions used when conducting our sperm selection procedure; these issues might include how long the sperm was stored after ejaculation and prior to sperm selection, what specific type of sperm selection procedure was employed, for how long and under what conditions was the sperm sample stored or incubated prior to fertilization and what was the quality of the donor oocytes to be used? it is logical to assume that the quality of the oocyte contributes a minimum of 50% to predictive nature of sperm DNA for pregnancy outcome
Summary
Predicting the success rate of human pregnancy using estimates of sperm DNA damage to date has been a very controversial issue, and that is no doubt further complicated by a plethora and permutation of techniques and procedures that vary in competency and delivery from laboratory to laboratory, all of which result in a fundamental lack of standardization. These variables include 1) the type of fertilization technique OPEN ACCESS. Given the wide range of variables that contribute to the success of an ART outcome, it is not surprising that this variability is reflected in the ability of sperm DNA fragmentation (SDF) to predict pregnancy outcome. This concept is well illustrated if we consider just one of the variables, such as sperm selection. In order to improve the predictive power of SDF it is important that we account and describe the precise conditions used when conducting our sperm selection procedure; these issues might include how long the sperm was stored after ejaculation and prior to sperm selection, what specific type of sperm selection procedure was employed, for how long and under what conditions was the sperm sample stored or incubated prior to fertilization and what was the quality of the donor oocytes to be used? it is logical to assume that the quality of the oocyte contributes a minimum of 50% to predictive nature of sperm DNA for pregnancy outcome
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