Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation

Abstract

ObjectiveThis study aimed to assess and compare sperm motility, concentration, and morphology recovery rates, before and after processing through sperm washing followed by swim-up or discontinuous density gradient centrifugation in normospermic individuals.MethodsFifty-eight semen samples were used in double intrauterine insemination procedures; 17 samples (group 1) were prepared with sperm washing followed by swim-up, and 41 (group 2) by discontinuous density gradient centrifugation. This prospective non-randomized study assessed seminal parameters before and after semen processing. A dependent t-test was used for the same technique to analyze seminal parameters before and after semen processing; an independent t-test was used to compare the results before and after processing for both techniques.ResultsThe two techniques produced decreases in sample concentration (sperm washing followed by swim-up: P<0.000006; discontinuous density gradient centrifugation: P=0.008457) and increases in motility and normal morphology sperm rates after processing. The difference in sperm motility between the two techniques was not statistically significant. Sperm washing followed by swim-up had better morphology recovery rates than discontinuous density gradient centrifugation (P=0.0095); and the density gradient group had better concentration recovery rates than the swim-up group (P=0.0027).ConclusionThe two methods successfully recovered the minimum sperm values needed to perform intrauterine insemination. Sperm washing followed by swim-up is indicated for semen with high sperm concentration and better morphology recovery rates. Discontinuous density gradient centrifugation produced improved concentration recovery rates.

Fluorescence-Based Ratiometric Analysis of Sperm Centrioles (FRAC) Finds Patient Age and Sperm Morphology Are Associated With Centriole Quality

“…These 31 patients were all undergoing infertility treatments but had varying diagnoses; their sperm phenotypes were classified as either eumorphic sperm (normal morphology) (n = 22) or teratospermia (poor morphology) based on Kruger's strict criteria (n = 9) (see patient details in Supplementary Table 1). Every sample was processed using gradient centrifugation, which resulted in two fractions: a pellet of dense sperm, generally regarded as higher quality, and used for intrauterine insemination, IUI (Karamahmutoglu et al, 2014;Fácio et al, 2016); and an accumulation of lighter sperm at the interface of the gradient phases, which has lower quality sperm (Branigan et al, 1999). The pellet sperm of every sample was collected (n = 31), and the interface from studies 2 and 3 were also collected (n = 19).…”

Reactive oxygen species in reproduction: harmful, essential or both?

“…Sperm washing by centrifugation and swim-up allow the separation of mobile and normal spermatozoa from abnormal spermatozoa and allows a better selection before IVF (Sakkas et al, 2000;Hammadeh et al, 2001;Henkel and Schill, 2003;Ren et al, 2004;Ricci et al, 2009;Jayaraman et al, 2012;Fácio et al, 2016;Santiago-Moreno et al, 2017;Kocaman, 2019;Rivera-Egea et al, 2019). Density gradient centrifugation was introduced to separate spermatozoa depending on chromosome packaging (Yamanaka et al, 2016) to select functional spermatozoa, while the swim-up method was designed to isolate highly motile sperm (Jayaraman et al, 2012;Beydola et al, 2014;Volpes et al, 2016).…”

“…However, other reports demonstrated that the duration of sperm-oocyte incubation does not significantly increase embryo viability (Swenson et al, 2000;Barraud-Lange et al, 2008). These conflicting reports may be due to the continuous amelioration of sperm washing techniques that minimizes sperm toxicity levels and culture medium composition that prevents oocyte and embryo contamination (Lane and Gardner, 2007;Morbeck et al, 2014;Fácio et al, 2016;Morbeck et al, 2017). Each IVF laboratory has its own manipulations techniques that can differently effect outcomes IVF.…”

Sperm preparation after freezing improves motile sperm count, motility, and viability in frozen-thawed sperm compared with sperm preparation before freezing-thawing process

“…Density gradient centrifugation (DGC) and swim-up are the two sperm preparation techniques most commonly used. In the literature, studies that choose either swim-up or DGC as a selection technique obtaining good results in both cases can be found [13][14][15]. The common point of these selection techniques is the fact that both remove seminal plasma present in fresh ejaculates and lead to a selection of a group of spermatozoa which exhibit better sperm features.…”