Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates

Gloria, Alessia; Carluccio, Augusto; Wegher, Laura; Robbe, Domenico; Contri, Alberto

doi:10.1186/s40104-016-0088-6

Cited by 25 publications

(33 citation statements)

References 40 publications

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“…To carry out simultaneous measurements of MMP and ROS on the same aliquots, a different fluorochrome would be needed; this possibility is currently being investigated. In contrast, Gloria et al (2016) were unable to detect a change in MMP levels between selected and unselected sperm samples using iodixanol as the separation medium.…”

Section: Discussionmentioning

confidence: 76%

“…In contrast, Gloria et al. () were unable to detect a change in MMP levels between selected and unselected sperm samples using iodixanol as the separation medium.…”

Section: Discussionmentioning

confidence: 92%

“…The effect of sperm selection using silane-silica-coated colloids immediately after semen collection on post-thaw sperm quality has not been investigated. Gloria et al (2016) were unable to show a beneficial effect when preparing "normal" quality bull semen on iodixanol, either as a density gradient (several layers of different densities) or as a single layer, although they could show an improvement in quality from poor quality ejaculates.…”

Section: Introductionmentioning

confidence: 95%

“…Gloria et al. () were unable to show a beneficial effect when preparing “normal” quality bull semen on iodixanol, either as a density gradient (several layers of different densities) or as a single layer, although they could show an improvement in quality from poor quality ejaculates.…”

Section: Introductionmentioning

confidence: 99%

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Effects of single layer centrifugation (SLC) on bull spermatozoa prior to freezing on post‐thaw semen characteristics

Nongbua

Johannisson

Edman³

et al. 2017

Reprod Domestic Animals

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Single layer centrifugation (SLC) has been shown to select the most robust spermatozoa from the ejaculate in several species. Here the effects of SLC prior to freezing on various parameters of frozen-thawed bovine sperm quality are reported. Semen from 8 bulls was layered on top of a species-specific colloid, Bovicoll. After centrifugation for 20 min at 300 g, the resulting sperm pellet was resuspended in OPTIXcell (IMV Technologies, l'Aigle, France); the SLC-selected sperm samples and uncentrifuged controls were frozen. On thawing, all sperm samples were analysed for membrane integrity, production of reactive oxygen species, mitochondrial membrane potential (MMP) and chromatin integrity. The SLC-treated samples had a higher percentage of live, superoxide-positive spermatozoa than uncentrifuged samples (27.9 ± 5.1% versus 21.7 ± 6.7%; p = .03). They had a higher proportion of spermatozoa with high mitochondrial membrane potential than uncentrifuged samples (55.9 ± 8.2% versus 40.5 ± 15.1%; p = .03) and also a lower proportion of spermatozoa with low mitochondrial membrane potential than non-treated samples (42.0 ± 8.5% versus 55.9 ± 14.4%; p = .04). No significant effects of treatment were found for membrane integrity or chromatin integrity. The effect of bull was significant on the proportions of dead, superoxide-positive spermatozoa and live, hydrogen peroxide-negative spermatozoa, as well as on membrane integrity, but it was not significant for mitochondrial membrane potential or chromatin integrity. These results suggest that SLC selects the most metabolically active bull spermatozoa from the rest of the population in normal ejaculates; the pattern of reactive oxygen species production may be different in SLC-selected spermatozoa compared to unselected samples.

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Section: Discussionmentioning

confidence: 76%

“…In contrast, Gloria et al. () were unable to detect a change in MMP levels between selected and unselected sperm samples using iodixanol as the separation medium.…”

Section: Discussionmentioning

confidence: 92%

Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Effects of single layer centrifugation (SLC) on bull spermatozoa prior to freezing on post‐thaw semen characteristics

Nongbua

Johannisson

Edman³

et al. 2017

Reprod Domestic Animals

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“…Other methods of sperm selection, such as the swimup method reduce the use of centrifugation, but this technique is still necessary to concentrate the sample. Studies report that high centrifugal forces often promote higher sperm recovery (WAITE et al, 2008;LEN et al, 2010;GLORIA et al, 2016) Some studies suggested that a reduction in the force and duration of centrifugation could reduce sperm damage and improve the sperm selection protocol using Percoll gradient (GUIMARÃES et al, 2014;KIM et al, 2013). However, these studies evaluated only the first centrifugation for sperm selection disregarding the second centrifugation for sperm concentration.…”

Section: Introductionmentioning

confidence: 99%

Use of a colloid to optimize centrifugation in the selection of bovine sperm for IVF

Gonçalves

Leivas

Míssio

et al. 2018

SCA

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This study aimed to evaluate the effect of the force and duration of centrifugation and the impact of cushioned centrifugation on sperm selection by Percoll gradient, on sperm quality and development kinetics of in vitro produced bovine embryos. Two experiments were performed. In Experiment I, a pool of semen was selected by Percoll gradients and the pellet was divided into four groups and distributed in a 2 × 2 factorial, with two forces (2200 × g or 9000 × g) and two durations (1 min or 3 min) of centrifugation. In Experiment II, semen was divided into two groups and selected by Percoll gradient with Cushion Fluid (CF) or without CF (Control) in the second centrifugation. The morphofunctionality, biochemical characteristics and fertilizing capacity of the selected sperms were evaluated. In addition, the development of the resulting bovine embryos was monitored for 48 h post-insemination. Duncan and Chi-square tests (P < 0.05) were used to compare the means. In Experiment I, there was a significant increase in sperm vigor (P < 0.05) after sperm selection in all treatments. The force and duration of centrifugation did not have any effect on sperm motility, vigor, and recovery rate among the different treatments (P > 0.05). In Experiment II, the recovery rate and reactive oxygen species (ROS) production in semen were similar among treatments (P > 0.05) although a higher ROS production was observed in the CF fertilization medium. Total fertilization rate was superior in the CF group (65.4 ± 5.3%) compared to that in Control (39.6 ± 4.9%). However, the normal fertilization and cleavage rate did not differ between the Control (94 ± 6.3% and 58.3 ± 8.3%) and CF (89 ± 7.1% and 75.0 ± 7.3%) groups. The reduction in the force and duration of centrifugation did not decrease the sperm recovery during selection by the Percoll gradient and the use of CF in the second centrifugation did not affect the normal fertilization and development of bovine IVF embryos up to 48 h. Key words: Bovine embryo. Cushion centrifugation. In vitro production. Percoll gradients. Sperm recovery. ResumoEsse estudo objetivou avaliar o efeito da força e duração da centrifugação, e o impacto da centrifugação amortecida na seleção espermática por gradientes de Percoll, na qualidade espermática e cinética do desenvolvimento de embriões bovinos produzidos in vitro. Dois experimentos foram realizados. No experimento I, um pool de sêmen foi selecionado por gradientes de Percoll e o pellet dividido em quatro grupos e distribuído em um fatorial 2 x 2, com duas forças (2200 e 9000 X g) e dois tempos (1 e 3 min) de centrifugação. No Experimento II, o sêmen foi dividido em dois grupos e selecionado com (CF) ou sem CushionFluid (Controle) na segunda centrifugação. A morfofuncionalidade, características bioquímicas e capacidade fecundante dos espermatozoides selecionados foram avaliadas. Além disso, o desenvolvimento dos embriões bovinos resultantes foi monitorado por 48 horas pós-inseminação. Os testes Duncan e Qui-quadrado (P<0,05) foram usados para ...

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Sperm Syringe: 3D Sorting Platform for Assisted Reproduction

Parast

O'Bryan

Nosrati

2022

Adv Materials Technologies

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sperm injection (ICSI), [3] in vitro fertilization (IVF), [4] and intrauterine insemination (IUI). [5] In ICSI, as the most invasive method, an individual sperm is selected and injected directly into an egg. In IVF or IUI, a subpopulation of pre-selected sperm is introduced close to the egg to achieve fertilization. [3][4][5] The quality of selected sperm is crucial to ART, contributing significantly to the treatment success rate, live-birth rate, and offspring health. [6] However, the current clinical sperm selection practices are highly manual, subjective, and prone to operator errors, resulting in a suboptimal ART success rate, stagnating at ≈33% per cycle over the past 30 years. [7,8] The inherent challenge is to mimic the highly parallelized and 3D selection process in vivo that enables a single-cell level sorting mechanism within a relatively short timeframe. [9] Specifically, the 3D folded structure of the female reproductive tract breaks down the initial semen volume (≈1-4 mL) into a few microliters per selection event within the folded epithelial of the fallopian tube. These subfractioning events can process hundreds of millions of sperm in just a few hours. [10] Current clinical practices, however, only provide a 2D bulk scale selection alternative.Density gradient centrifugation (DGC) [11] and swim-up [12,13] are the most commonly used methods for human sperm selection in fertility clinics. In swim-up assay (SU), highly motile sperm are selected based on their ability to swim upward from the raw semen sediment into a fresh sperm media, where they are collected after ≈60 min. This method is not applicable in the case of asthenozoospermia (poor sperm motility). [14] In DGC, human sperm are separated based on their density by being forced to cross a viscosity gradient via centrifugation force that could cause sperm DNA damage and alter sperm morphology and function. [11,15] The sperm preparation method that is used in clinics is mainly chosen based on the count and motility of sperm in the human semen sample. [9] Both SU and DGC are time consuming (30-60 min), require multiple preparation steps, depend on the clinician expertise, and differ significantly from the natural selection process in vivo. [16] Moreover, the relatively long processing time of these methods and the low percentage of recovered sperm in the swim-up method (less than 20%) have considerably restricted the clinical workflow, especially Selection of high-quality sperm is crucial to assisted reproduction. However, conventional clinical methods for sperm selection are manual and prone to operator errors. This article presents 'sperm syringe', a scalable technology that mimics the highly parallelized 3D selection process in vivo via a 3D network of 560 microchannels to select high-quality sperm. Sperm syringe retrieves more than 41% of healthy sperm from the initial sperm sample in under 15 min, providing a sufficient volume (≈500 µL) and number (1,600,000) of high-quality sperm for fertility treatments. Experiments with bull and human sper...

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Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates

Cited by 25 publications

References 40 publications

Effects of single layer centrifugation (SLC) on bull spermatozoa prior to freezing on post‐thaw semen characteristics

Effects of single layer centrifugation (SLC) on bull spermatozoa prior to freezing on post‐thaw semen characteristics

Use of a colloid to optimize centrifugation in the selection of bovine sperm for IVF

Sperm Syringe: 3D Sorting Platform for Assisted Reproduction

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