Isolation of spermatozoa with low levels of fragmented DNA with the use of flow cytometry and sorting

Ribeiro, Sofia; Sartorius, Gideon; Pletscher, Flurina; Geyter, Maria De; Zhang, Hong; Geyter, Christian De

doi:10.1016/j.fertnstert.2013.05.030

Cited by 23 publications

(27 citation statements)

References 62 publications

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“…In this paper (Ribeiro et al., ) was found that both dead and apoptotic spermatozoa are labeled; while the dye does not stain the viable spermatozoa. Compared with the conventional swim‐up method, the sorted viable population showed a significantly reduced number of spermatozoa with fragmented DNA (Ribeiro et al., ). This finding also shed light on the origin of DNA fragmentation in human spermatozoa.…”

Section: The Membrane: Increased Permeability Phospholipid Transposimentioning

confidence: 81%

“…The combination of YoPro‐1 and PI has been extensively used in the laboratory of the authors, and has proven to show changes in the membrane at a much earlier stage than the combination SYBR‐14/PI (Gallardo Bolanos et al., ). The YoPro‐1/PI assay show better correlation with motility, and, a human study, demonstrate that YoPro‐1 negative spermatozoa also show better DNA integrity (Ribeiro et al., ). In this paper (Ribeiro et al., ) was found that both dead and apoptotic spermatozoa are labeled; while the dye does not stain the viable spermatozoa.…”

Section: The Membrane: Increased Permeability Phospholipid Transposimentioning

confidence: 97%

“…The YoPro‐1/PI assay show better correlation with motility, and, a human study, demonstrate that YoPro‐1 negative spermatozoa also show better DNA integrity (Ribeiro et al., ). In this paper (Ribeiro et al., ) was found that both dead and apoptotic spermatozoa are labeled; while the dye does not stain the viable spermatozoa. Compared with the conventional swim‐up method, the sorted viable population showed a significantly reduced number of spermatozoa with fragmented DNA (Ribeiro et al., ).…”

Section: The Membrane: Increased Permeability Phospholipid Transposimentioning

confidence: 97%

See 2 more Smart Citations

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Peña

Rodriguez

Gil

et al. 2018

Reprod Domestic Animals

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Contents Flow cytometry is increasingly used in research and also in clinical andrology. Recent developments in instrumentation, availability of probes and bioinformatics expand the possibilities of flow cytometry well beyond the classical two parametric analyses in use. In this paper, an overview of recent developments in flow cytometry will be presented under the perspective of the authors; aspects such a multicolor assays and computational cytometry will be discussed as well.

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Section: The Membrane: Increased Permeability Phospholipid Transposimentioning

confidence: 81%

Section: The Membrane: Increased Permeability Phospholipid Transposimentioning

confidence: 97%

Section: The Membrane: Increased Permeability Phospholipid Transposimentioning

confidence: 97%

See 1 more Smart Citation

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Peña

Rodriguez

Gil

et al. 2018

Reprod Domestic Animals

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“…Using FCM, various sperm features were measured such as sperm concentration, 31 sperm viability and acrosomal integrity, 32 33 34 mitochondrial function, 35 and DNA integrity. 36 37 38 Fluorescence-activated cell sorting applied in FCM is used for sorting sperm population with defined properties, e.g., with a reduced number of spermatozoa with fragmented DNA 39 and a selection of spermatogenic cells from testicular biopsies. 40 …”

Section: Discussionmentioning

confidence: 99%

Evaluation of the expression of sperm proteins in normozoospermic and asthenozoospermic men using monoclonal antibodies

Capková¹,

Kubátová²,

Děd³

et al. 2016

Asian J Androl

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Recent studies have shown that infertility affects estimated 15% of all couples. Male infertility is the primary or contributory cause in 60% of these cases. Consequently, the application of assisted reproduction is increasing. These methods could benefit from an extended evaluation of sperm quality. For this reason, we analyzed sperm proteins from 30 men with normal spermiograms and 30 men with asthenozoospermia. Ejaculates of both groups were tested by flow cytometry (FCM) and fluorescence with a set of well-characterized anti-human sperm Hs-monoclonal antibodies (MoAbs), which were generated in our laboratory. No statistically significant differences were found between normospermics and asthenospermics in the expression of the sperm surface protein clusterin, evaluated with Hs-3 MoAb, and semenogelin, evaluated with Hs-9 MoAb. However, FCM revealed quantitative differences in the acrosomal proteins between normozoospermic and asthenozoospermic men, namely, in glyceraldehyde-3-phosphate dehydrogenase, evaluated with Hs-8 MoAb, valosin-containing protein, evaluated with Hs-14 MoAb, and ATP synthase (cAMP-dependent protein kinase II, PRKAR2A), evaluated with MoAb Hs-36. Asthenozoospermic men displayed a highly reduced expression of intra-acrosomal proteins, with a likely decrease in sperm quality, and thus a negative impact on successful reproduction. Asthenozoospermia seems to be a complex disorder involving intra-acrosomal proteins.

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“…The researchers behind the study showed that it is possible to separate the dead spermatozoa and the spermatozoa with increased amount of DNA fragmentation from the normal spermatozoa. This makes it possible to optimize the sperm sample before fertility treatment like ICSI is initiated (Ribeiro et al ., ). However, recent research has shown that methods where ICSI is optimized remains controversial (Tavalaee et al ., ; Troya & Zorrilla, ).…”

Section: Historical Overviewmentioning

confidence: 97%

DNA fragmentation in spermatozoa: a historical review

Rex

Aagaard²,

Fedder

2017

Andrology

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SummarySperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated. In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage. The present decade continues within this research area. Some of the more novel methods recently submerging are sorting of cells with increased DNA fragmentation and hyaluronic acid (HA) binding techniques. The clinical value of these tests remains to be elucidated. In spite of half a century of research within the area, this analysis is not routinely implemented into the fertility clinics. The underlying causes are multiple. The abundance of methods has impeded the need for a clinical significant threshold. One of the most promising methods was commercialized in 2005 and has been reserved for larger licensed laboratories. Myriads of reviews and meta‐analyses on studies using different assays for analysis of DNA fragmentation, different clinical Artificial Reproductive Treatments (ART), different definitions of successful ART outcome and small patient cohorts have been published. Although the area of DNA fragmentation in spermatozoa is highly relevant in the fertility clinics, the need for further studies focusing on standardization of the methods and clinical implementation persists.

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Isolation of spermatozoa with low levels of fragmented DNA with the use of flow cytometry and sorting

Cited by 23 publications

References 62 publications

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Evaluation of the expression of sperm proteins in normozoospermic and asthenozoospermic men using monoclonal antibodies

DNA fragmentation in spermatozoa: a historical review

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