Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters

Chiamchanya, Charoenchai; Kaewnoonual, Nattpawit; Visutakul, Pachara; Manochantr, Sirikul; Chaiya, Jirattikan

doi:10.1038/aja.2009.60

Cited by 24 publications

(28 citation statements)

References 19 publications

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“…Xue et al (2014) compared swim-up and discontinuous density gradient centrifugation in patients with teratozoospermia, and their results suggested that enrichment with normal morphology spermatozoa with intact DNA could be achieved through swim-up or discontinuous density gradient centrifugation, when compared to unprocessed semen. Other authors have reported similar results (Ng et al ., 1992; Hammadeh et al ., 2001; Chiamchanya et al ., 2010). Furthermore, Xue et al .…”

Section: Discussionmentioning

confidence: 99%

Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation

Facio¹,

Previato²,

Machado-Paula³

et al. 2016

JBRA Assisted Reproduction

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ObjectiveThis study aimed to assess and compare sperm motility, concentration, and morphology recovery rates, before and after processing through sperm washing followed by swim-up or discontinuous density gradient centrifugation in normospermic individuals.MethodsFifty-eight semen samples were used in double intrauterine insemination procedures; 17 samples (group 1) were prepared with sperm washing followed by swim-up, and 41 (group 2) by discontinuous density gradient centrifugation. This prospective non-randomized study assessed seminal parameters before and after semen processing. A dependent t-test was used for the same technique to analyze seminal parameters before and after semen processing; an independent t-test was used to compare the results before and after processing for both techniques.ResultsThe two techniques produced decreases in sample concentration (sperm washing followed by swim-up: P<0.000006; discontinuous density gradient centrifugation: P=0.008457) and increases in motility and normal morphology sperm rates after processing. The difference in sperm motility between the two techniques was not statistically significant. Sperm washing followed by swim-up had better morphology recovery rates than discontinuous density gradient centrifugation (P=0.0095); and the density gradient group had better concentration recovery rates than the swim-up group (P=0.0027).ConclusionThe two methods successfully recovered the minimum sperm values needed to perform intrauterine insemination. Sperm washing followed by swim-up is indicated for semen with high sperm concentration and better morphology recovery rates. Discontinuous density gradient centrifugation produced improved concentration recovery rates.

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Section: Discussionmentioning

confidence: 99%

Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation

Facio¹,

Previato²,

Machado-Paula³

et al. 2016

JBRA Assisted Reproduction

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show abstract

“…The application of assisted reproduction procedures to select mature sperm with nondamaged DNA is important to minimize the generation of mutations and their transmission to offspring (Leduc et al, 2008). It has been described that DGC results in a marked reduction in DNA fragmentation (Sakkas et al, 2000; Morell et al, 2004; Chiamchanya et al, 2010). Our results also indicated a marked reduction in DNA fragmentation when comparing native with DGC sperm (Table 1).…”

Section: Discussionmentioning

confidence: 99%

Protamine/DNA Ratios and DNA Damage in Native and Density Gradient Centrifuged Sperm From Infertile Patients

Castillo

Lee

Mateo

et al. 2011

Journal of Andrology

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Protamines are the major nuclear proteins condensing DNA in the sperm nucleus. One of their proposed functions is the protection of the genetic message delivered by the sperm. To date, evidence of their involvement in DNA protection has been obtained by correlating the protamine P1/P2 ratio, protamine concentrations, or chromomycin A3 staining with DNA fragmentation. However, a correlation of the absolute protamine/DNA content with the DNA fragmentation in sperm from the same infertile patients as assessed with the comet assay has not been studied. Protamine/DNA ratios were calculated after protamine and DNA extraction, electrophoresis, and gel quantification of the protamines and DNA quantification in the sperm samples of 66 infertile patients before (native sample) and after a 2-step discontinuous PureSperm density gradient centrifuged (DGC) selection of the sperm. DNA fragmentation was assessed using the alkaline comet assay. In DGC sperm, the total protamine/DNA, P1/DNA, and P2/DNA ratios all correlated inversely with DNA damage in sperm from infertile patients. The detection of this inverse correlation between protamine/DNA ratios and DNA damage in DGC sperm adds support to the hypothesis that defective protamination is related to DNA damage in the clinically relevant subpopulation of sperm from infertile men.

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“…Chiamchanya et al [25] compared the sperm quality Figure 2. Effect of incubation temperatures and time on the motility and DNA fragmentation index (%DFI) of spermatozoa in raw semen (not frozen and thawed).…”

Section: Discussionmentioning

confidence: 99%

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Matsuura¹,

Takeuchi²,

Yoshida³

2010

Asian J Androl

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Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay (SCSA) parameters, that is, DNA fragmentation index (%DFI) and high DNA stainability (%HDS), were assessed on the fresh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation (DGC) was followed by swimup treatment in comparison with DGC alone (P < 0.01). Although the %DFI increased in a time-dependent manner with incubation both at room temperature (RT) and at 37ºC in air, the %DFI after 24 h at RT was significantly lower than that at 37ºC (P < 0.05). Incubation with 5% CO2 was effective in maintaining sperm motility (P < 0.01); however, it induced further elevation of %DFI (P < 0.001). Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.

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Comparative study of the effects of three semen preparation media on semen analysis, DNA damage and protamine deficiency, and the correlation between DNA integrity and sperm parameters

Cited by 24 publications

References 19 publications

Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation

Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation

Protamine/DNA Ratios and DNA Damage in Native and Density Gradient Centrifuged Sperm From Infertile Patients

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

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