Removal of DNA-fragmented spermatozoa using flow cytometry and sorting does not improve the outcome of intracytoplasmic sperm injection

Geyter, Christian De; Gobrecht-Keller, Ursula; Ahler, Astrid; Fischer, Manuel

doi:10.1007/s10815-019-01571-1

Cited by 9 publications

(6 citation statements)

References 29 publications

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“…The oxidative attack on sperm DNA can lead to the formation of oxidised bases, making the DNA strand prone to fragmentation. These spermatozoa are essentially cryptic in terms of SDF detection, waiting 'under the surface', ultimately to be detected, depending on the degree of damage imposed, for instance, by ex-vivo manipulation or iatrogenic damage before use in ART.Indeed, it has been shown that the decrease in SDF rates seen after the use of sperm selection techniques for ART does not necessarily translate to improvements in pregnancy rates(De Geyter et al, 2019;Gosálvez et al, 2013). Moreover, a few double-strand DNA breaks are sufficient to delay cell cycle progression(van den Berg et al, 2018).…”

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confidence: 99%

Sperm DNA fragmentation testing: Summary evidence and clinical practice recommendations

et al. 2020

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confidence: 99%

Sperm DNA fragmentation testing: Summary evidence and clinical practice recommendations

et al. 2020

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“…Until recently, FACS sorting procedure of spermatozoa was mainly used for sex sorting. However, a few recent studies (Ribeiro et al, 2013;De Geyter et al, 2019) showed that using a YO-PRO-1 staining and FACS sorting the proportion of DNA-fragmented spermatozoa in human semen samples can be reduced to 5% as a result of apoptotic cell removal. In general, fragmentation of nuclear DNA is one of the main features of apoptosis (Ribeiro et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…However, the reproductive outcomes regarding this method are unknown. As an alternative to MACS, FACS sorting of YO-PRO-1 positive cells were recently used to remove apoptotic cells and thus reduced the number of DNAfragmented spermatozoa in order to improve the outcomes in assisted reproduction (Ribeiro et al, 2013;De Geyter et al, 2019). Thus, beside the sperm sexing FACS sorting might be used also to improve semen quality.…”

Section: Introductionmentioning

confidence: 99%

Optimization of Facs Sorting for the Improvement of Livestock Semen Quality

Vasicek,

Svoradova,

Balazi

et al. 2021

J microb biotech food sci

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Fluorescence-activated cell sorting (FACS) of spermatozoa was mainly used for sex sorting. Recently, FACS has been used to eliminate DNA-damaged human spermatozoa using YO-PRO-1 staining. Fragmentation of sperm DNA is considered as one of the reasons of male infertility. Moreover, YO-PRO-1 can effectively mark apoptotic as well as dead spermatozoa. Till now, only two FACS instruments were used for the spermatozoa sorting. However, both of them are not more commercially available from different reasons. In this study, we used novel FACSMelody Cell Sorter for the elimination of apoptotic and dead cells from the rabbit and ram semen samples in order to improve their overall quality. Briefly, semen samples were stained using YO-PRO-1 dye (apoptotic and dead cells) and/or propidium iodide (PI; only dead cells). Three different sorting experiments were performed: E1 – YO-PRO-1 and PI stained rabbit sperm cells were sorted into the tubes containing 1 ml of PBS; E2 – PI stained rabbit sperm cells were sorted into tubes that were washed with FBS prior adding PBS; and E3 – YO-PRO-1 and PI stained ram sperm cells were sorted into tubes washed with FBS prior adding PBS. As a sheath fluid sterile PBS was used. All samples, control (before sorting), negatively and positively sorted fractions, were analysed using CASA for motility assessment. Moreover, all sorted samples were re-stained with PI for viability assessment. In conclusion, elimination of dead (PI+) sperm from rabbit samples might improve their quality, since their progressive motility increased significantly (P<0.001) after sorting from 40 to 65%. However, ram spermatozoa seem to be sensitive to sorting procedure thus further optimisation of this procedure is required.

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“…Sperm negatively selected via Annexin-V FACS have been shown to possess intact chromatin [ 107 ]. Additionally, in a human clinical trial, spermatozoa from the population that did not bind Annexin-V during FACS were used for ICSI resulted in improved pregnancy and live birth rates, as well as reduced miscarriage rates over embryos generated with sperm selected from SU [ 112 ].…”

Section: Sorting Semen: Significance and Methodsmentioning

confidence: 99%

An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section II

Orsolini

Meyers

Dini

2021

Animals

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As the use of assisted reproductive technologies (ART) and in vitro embryo production (IVP) expand in the equine industry, it has become necessary to further our understanding of available semen selection techniques. This segment of our two-section review will focus on the selection of spermatozoa based on quality and sex for equine intracytoplasmic sperm injection (ICSI), as well as current and future developments in sperm sorting technologies. Ultimately, novel methods of semen selection will be assessed based on their efficacy in other species and their relevance and future application towards ARTs in the horse.

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Removal of DNA-fragmented spermatozoa using flow cytometry and sorting does not improve the outcome of intracytoplasmic sperm injection

Cited by 9 publications

References 29 publications

Sperm DNA fragmentation testing: Summary evidence and clinical practice recommendations

Sperm DNA fragmentation testing: Summary evidence and clinical practice recommendations

Optimization of Facs Sorting for the Improvement of Livestock Semen Quality

An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section II

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