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

Raad, Georges; Bakos, Hassan W.; Bazzi, Marwa; Mourad, Youmna; Fakih, Fadi; Shayya, Salman; Mchantaf, Lama; Fakih, Chadi

doi:10.1111/andr.13038

Cited by 24 publications

(17 citation statements)

References 101 publications

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“…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.…”

Section: Discussionmentioning

confidence: 99%

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

Huang

et al. 2021

Front. Cell Dev. Biol.

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In this paper, on the basis of the differences in the hydrogen ion concentration (pH) of the diluent dairy goat semen on X/Y sperm motility, an X/Y sperm enrichment study was conducted to establish a simple and effective method for gender control in dairy goats. Dairy goat semen was diluted using different pH dilutions and was incubated. Then, the X/Y sperm ratio in the isolated upper sperm was determined using the double TaqMan qPCR method. The internal pH change pattern of sperm cells at different pH dilutions was measured using BCECF-AM probe, and the functional parameters of the isolated sperm were tested with the corresponding kit. Next, an in vitro fertilization test was conducted using isolated spermatozoa and oocytes to determine their fertilization rates, the percentages of female embryos, and the expression of genes related to developing potentially fertilized embryos. Results showed that the percentages of the X sperm cells in the upper sperm layer were 67.24% ± 2.61% at sperm dilution pH of 6.2 and 30.45% ± 1.03% at sperm dilution pH of 7.4, which was significantly different from 52.35% ± 1.72% of the control group (pH 6.8) (P < 0.01). Results also showed that there is a relationship between the external pHo and internal pHi of sperm cells. Furthermore, the percentages of female embryos after the in vitro fertilization of the isolated upper sperm with mature oocytes at pH 6.2 and 7.4 were 66.67% ± 0.05 and 29.73% ± 0.04%, respectively, compared with 48.57% ± 0.02% in the control group (pH 6.8). Highly significant differences occurred between groups (P < 0.01). Additionally, no significant difference was observed during the expression of genes related to embryonic development between the blastocysts formed from sperm isolated by changing the pH of the diluent and the control sperm (P > 0.05). Therefore, this study successfully established a simple and effective method for enriched X/Y sperms from dairy goats, which is important for regulating the desired sex progeny during dairy goat breeding and for guiding dairy goat production.

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Section: Discussionmentioning

confidence: 99%

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

Huang

et al. 2021

Front. Cell Dev. Biol.

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“…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.…”

Section: Classic Methodsmentioning

confidence: 99%

Sperm Selection for ICSI: Do We Have a Winner?

Baldini

Ferri

Baldini

et al. 2021

Cells

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In assisted reproductive technology (ART), the aim of sperm cells’ preparation is to select competent spermatozoa with the highest fertilization potential and in this context, the intracytoplasmic sperm injection (ICSI) represents the most applied technique for fertilization. This makes the process of identifying the perfect spermatozoa extremely important. A number of methods have now been developed to mimic some of the natural selection processes that exist in the female reproductive tract. Although many studies have been conducted to identify the election technique, many doubts and disagreements still remain. In this review, we will discuss all the sperm cell selection techniques currently available for ICSI, starting from the most basic methodologies and continuing with those techniques suitable for sperm cells with reduced motility. Furthermore, different techniques that exploit some sperm membrane characteristics and the most advanced strategy for sperm selection based on microfluidics, will be examined. Finally, a new sperm selection method based on a micro swim-up directly on the ICSI dish will be analyzed. Eventually, advantages and disadvantages of each technique will be debated, trying to draw reasonable conclusions on their efficacy in order to establish the gold standard method.

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“…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].…”

Section: Microfluidics and Potential Use In Spermatozoa Isolation From Noa Testicular Samplesmentioning

confidence: 99%

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

Smith

Cantatore²,

Ohl

2021

JCM

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Intracytoplasmic sperm injection (ICSI) has allowed reproduction options through assisted reproductive technologies (ARTs) for men with no spermatozoa within the ejaculate (azoospermia). In men with non-obstructive azoospermia (NOA), the options for spermatozoa retrieval are testicular sperm extraction (TESE), testicular sperm aspiration (TESA), or micro-surgical sperm extraction (microTESE). At the initial time of spermatozoa removal from the testis, spermatozoa are immobile. Independent of the means of spermatozoa retrieval, the subsequent steps of removing spermatozoa from seminiferous tubules, determining spermatozoa viability, identifying enough spermatozoa for oocyte injections, and isolating viable spermatozoa for injection are currently performed manually by laboratory microscopic dissection and collection. These laboratory techniques are highly labor-intensive, with yield unknown, have an unpredictable efficiency and/or success rate, and are subject to inter-laboratory personnel and intra-laboratory variability. Here, we consider the potential utility, benefits, and shortcomings of developing technologies such as motility induction/stimulants, microfluidics, dielectrophoresis, and cell sorting as andrological laboratory add-ons to reduce the technical burdens and variabilities in viable spermatozoa isolation from testicular samples in men with NOA.

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

Cited by 24 publications

References 101 publications

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

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

Sperm Selection for ICSI: Do We Have a Winner?

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

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