Sperm Chromatin Structure Assay (SCSA®): 30 Years of Experience with the SCSA®

Evenson, Donald P.

doi:10.1007/978-1-4419-6857-9_9

Cited by 31 publications

(37 citation statements)

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“…4 C). The HDS fraction contains sperm with an increased histone/protamine ratio and the presence of unprocessed protamines [ 22 ] and greater round morphology [ 69 ]. The HDS values were higher in treated WT compared to nontreated WT spermatozoa ( P < 0.05).…”

Section: Resultsmentioning

confidence: 99%

“…Altered chromatin structure measured by the susceptibility of sperm DNA to acid-induced denaturation at sites of existing DNA strand breaks [ 66 ] was assessed with the sperm chromatin structure assay (SCSA) [ 56 , 67 – 69 ]. A minimum of 10 000 of AO-labeled spermatozoa were analyzed for each sample using a MACSQuant Analyzer flow cytometer (Miltenyi Biotec, Inc., Auburn, CA) equipped with a 585/625-nm filter and a laser tuned to 488-nm line excitation.…”

Section: Methodsmentioning

confidence: 99%

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Absence of Peroxiredoxin 6 Amplifies the Effect of Oxidant Stress on Mobility and SCSA/CMA3 Defined Chromatin Quality and Impairs Fertilizing Ability of Mouse Spermatozoa1

Ozkosem

Feinstein

Fisher

et al. 2016

Biology of Reproduction

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Oxidative stress, the imbalance between reactive oxygen species production and antioxidant defenses, is associated with male infertility. Peroxiredoxins (PRDXs) are antioxidant enzymes with a wide distribution in spermatozoa. PRDX6 is highly abundant and located in all subcellular compartments of the spermatozoon. Infertile men have lower levels of sperm PRDX6 associated with low sperm motility and high DNA damage. In order to better understand the role of PRDX6 in male reproduction, the aim of this study was to elucidate the impact of the lack of PRDX6 on male mouse fertility. Spermatozoa lacking PRDX6 showed significantly increased levels of cellular oxidative damage evidenced by high levels of lipid peroxidation, 8-hydroxy-deoxyguanosine (DNA oxidation), and protein oxidation (S-glutathionylation and carbonylation), lower sperm chromatin quality (high DNA fragmentation and low DNA compaction, due to low levels of protamination and a high percentage of free thiols), along with decreased sperm motility and impairment of capacitation as compared with wild-type (WT) spermatozoa. These manifestations of damage are exacerbated by tert-butyl hydroperoxide treatment in vivo. While WT males partially recovered the quality of their spermatozoa (in terms of motility and sperm DNA integrity), Prdx6−/− males showed higher levels of sperm damage (lower motility and chromatin integrity) 6 mo after the end of treatment. In conclusion, Prdx6−/− males are more vulnerable to oxidative stress than WT males, resulting in impairment of sperm quality and ability to fertilize the oocyte, compatible with the subfertility phenotype observed in these knockout mice.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Absence of Peroxiredoxin 6 Amplifies the Effect of Oxidant Stress on Mobility and SCSA/CMA3 Defined Chromatin Quality and Impairs Fertilizing Ability of Mouse Spermatozoa1

Ozkosem

Feinstein

Fisher

et al. 2016

Biology of Reproduction

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“…Fertility clinics were to ship sperm samples to these larger diagnostic centres in order to obtain a SCSA‐DFI value (Evenson, ). As it is encouraged that DFI is determined using the commercial SCSAsoft ® software the SCSA analysis have primarily been restricted to the larger licensed diagnostic laboratories (Evenson, ). In order for the analysis to be implemented into the individual fertility clinics, an investment in a flow cytometer is required.…”

Section: Discussionmentioning

confidence: 99%

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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“…Sperm DNA damage was detected using the Sperm Chromatin structure assay (SCSA, CellPro Biotech Co., Ltd., Ningbo, China) according to the manufacturer's protocol and expressed as sperm DNA fragmentation index (DFI). The SCSA is the current accepted gold standard for sperm DNA damage testing (Evenson, ). It was suggested to reduce DFI value by change of lifestyle and/or medical intervention when DFI > 25% in human (Evenson, ).…”

Section: Methodsmentioning

confidence: 99%

“…The SCSA is the current accepted gold standard for sperm DNA damage testing (Evenson, ). It was suggested to reduce DFI value by change of lifestyle and/or medical intervention when DFI > 25% in human (Evenson, ). Therefore, we divided semen samples into two groups (DFI < 25% and ≥25%) according to the threshold value of DFI (25%).…”

Section: Methodsmentioning

confidence: 99%

Methylation levels ofIGF2andKCNQ1in spermatozoa from infertile men are associated with sperm DNA damage

Pan

et al. 2019

Andrologia

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Abnormal imprinted genes methylation in spermatozoa has been shown to be associated with subfertility. However, the relationship between sperm DNA damage and specific imprinted genes methylation remains unclear. In this study, DNA methylation levels were determined at seven imprinted genes loci (H19, INS‐IGF2, KCNQ1, MEG3, MEST, PEG3 and SNRPN) in 66 semen samples using the MSRE‐qPCR method. The semen samples were divided into two groups according to the threshold value (25%) of DNA fragmentation index (DFI). We found that the mean methylation level at IGF2 (cg17037101) in the group with DFI ≥ 25% was lower than that in the group with DFI < 25% (13.7 ± 3% vs. 31.5 ± 5.3%, p = 0.0053). However, the methylation levels of other CpGs did not differ from the imprinted genes. Correlation analysis of DFI with the methylation levels of imprinted genes demonstrated that the IGF2 (cg17037101) methylation level was negatively correlated with sperm DFI (r = −0.448, p = 0.0038), and the KCNQ1 (cg24932449) methylation level was positively correlated with sperm DFI (r = 0.354, p = 0.0273). Our results suggest that the aberrant methylation of IGF2 and KCNQ1 genes may be associated with sperm DNA damage.

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Sperm Chromatin Structure Assay (SCSA®): 30 Years of Experience with the SCSA®

Cited by 31 publications

References 50 publications

Absence of Peroxiredoxin 6 Amplifies the Effect of Oxidant Stress on Mobility and SCSA/CMA3 Defined Chromatin Quality and Impairs Fertilizing Ability of Mouse Spermatozoa1

Absence of Peroxiredoxin 6 Amplifies the Effect of Oxidant Stress on Mobility and SCSA/CMA3 Defined Chromatin Quality and Impairs Fertilizing Ability of Mouse Spermatozoa1

DNA fragmentation in spermatozoa: a historical review

Methylation levels ofIGF2andKCNQ1in spermatozoa from infertile men are associated with sperm DNA damage

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