Differential impact of four sperm preparation techniques on sperm motility, morphology, DNA fragmentation, acrosome status, oxidative stress, and mitochondrial activity: A prospective study

Abstract

Background: Suboptimal human semen handling in vitro may induce sperm damage.However, the effects of semen swim-up, pellet swim-up, density gradient, and density gradient followed by SU on sperm motility, morphology, DNA fragmentation, acrosome reaction, intracellular reactive oxygen species, and mitochondrial activity were not fully understood.Objectives: To study the impact of four sperm preparation techniques on sperm functional parameters. Materials and methods:This study was conducted on 60 infertile men with a minimum sperm concentration of 20 × 10 6 /ml and total sperm motility of ≥30%. Each raw semen sample was divided into four aliquots. Each aliquot was prepared by one of the tested techniques. Various sperm characteristics were assessed before and after sperm preparation.Results: Density gradient and density gradient followed by SU resulted in significantly higher DNA fragmentation percentages compared with semen swim-up (p < 0.001 and p < 0.001, respectively) and pellet swim-up (p < 0.001 and p < 0.001, respectively). Significantly higher percentages of spermatozoa with intact acrosome were detected in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen. The percentage of reactive oxygen species-positive spermatozoa was significantly higher after pellet swim-up (p < 0.001), density gradient (p < 0.001), and density gradient followed by SU (p < 0.001) than raw semen. In addition, the percentages of 100% stained midpiece (active mitochondria) were significantly higher in semen swim-up (p < 0.001) and pellet swim-up (p < 0.001) compared with raw semen. Discussion and conclusion:To the best of our knowledge, this is the first report comparing the impact of these techniques on various sperm functional parameters.Semen swim-up was more effective than density gradient in selecting better spermatozoa in terms of DNA integrity, reactive oxygen species levels, acrosome status, and

Effects of Diluent pH on Enrichment and Performance of Dairy Goat X/Y Sperm

“…The sperm acrosome contains many hydrolytic enzymes that are closely related to fertilization, and acrosome integrity facilitates the fertilization process (van der Horst, 2021). Several mitochondria are distributed at the neck of the sperm, which provides energy for sperm metabolism and movement, and participate in several fertilization processes, such as sperm hyperactivation, capacitation, and acrosome reaction (Nesci et al, 2020;Raad et al, 2021). Additionally, ROS is the main free radical in sperm cells.…”

Sperm Selection for ICSI: Do We Have a Winner?

“…Moreover, the latter is less efficient than SU in selecting sperm cells with intact DNA [ 66 ]. While these theories have been reinforced by more recent studies [ 67 ], in contrast, other authors have shown that DGC is better than SU to select a higher percentage of capacitated sperm [ 68 ] with hyper-activated motion even in cases of subfertility [ 69 ]. Other scientists suggest that neither method provides a cell population with acceptable levels of intact DNA.…”

Microfluidic Systems for Isolation of Spermatozoa from Testicular Specimens of Non-Obstructive Azoospermic Men: Does/Can It Improve Sperm Yield?

“…The technical parameters of the device were designed to optimize the isolation of motile human spermatozoa with the inflow channel (semen; 100 µm × 50 µm; width × depth), inflow channel (media; 300 µm × 50 µm), common midchannel (laminar flow; 500 µm × 50 µm × 100 mm length). Centrifugation can negatively influence sperm motility [32], mitochondrial function [33], intact acrosomal status [33], and DNA integrity [34]. Using the microfluidic laminar flow and inertia spermatozoa isolation, it was demonstrated that isolated motile spermatozoa had significantly less DNA damage compared to processing sperm with centrifugation, density gradient and centrifugation, and swim-up of overlaid semen [35][36][37].…”