The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies

Sakkas, Denny; Manicardi, Gian Carlo; Tomlinson, Mathew; Mandrioli, Mauro; Bizzaro, Davide; Bianchi, Patrizia; Bianchi, Umberto

doi:10.1093/humrep/15.5.1112

Cited by 214 publications

(140 citation statements)

References 23 publications

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“…While one study showed that the DGC is more effective than SUP in reducing the percentage of sperm with DNA damage [25], another one, in contrast, demonstrated that SUP recovers lower percentage of sperm with fragmented DNA as compared to DGC [37]. Enciso et al [11] showed that SUP and DGC are equally efficient in eliminating spermatozoa containing double-strand DNA damage and sperm with highly damaged DNA, as characterized by the presence of both single-and double-strand DNA breaks.…”

Section: Discussionmentioning

confidence: 99%

“…Several studies addressed whether there was any differences between these two methods regarding sperm motility and concentration after semen preparation and the outcomes of intrauterine insemination [1,6,10,25,35], in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) [9,[18][19][20]22,32,33] However the results are controversial.…”

Section: Introductionmentioning

confidence: 99%

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Intracytoplasmic morphologically selected sperm injection outcomes: the role of sperm preparation techniques

Borges

Setti

Vingris

et al. 2013

J Assist Reprod Genet

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Purpose To compare the results of intracytoplasmic morphologically selected sperm injection (IMSI) between cycles in which the swim-up (SUP) or the density gradient centrifugation (DGC) techniques were used for sperm preparation. Methods We evaluated 70 IMSI cycles performed in women with age ≤ 37 years, undergoing IMSI as result of male factor. The couples were divided into two groups: DGC group (n=26) and SUP group (n=44). The groups were compared with regard to IMSI outcomes. Results There were no significant differences between SUP and DGC groups regarding the number of follicles, oocytes, mature oocytes, oocyte yield and mature oocyte rate. Fertilization rate and high-quality embryos rate on day 5 of development were similar between SUP and DGC groups. Implantation, pregnancy and miscarriage rates were not statistically different between SUP and DGC groups (28.8 vs 33.3 %, 46.2 vs 57.1 % and 8.3 vs 4.2 %, respectively). Conclusions Both the SUP and the DGC techniques recover improved sperm fractions and result in similar IMSI outcomes. Further randomized trials analyzing both the quality of sperm through MSOME and the IMSI outcomes are needed to elucidate the role of sperm preparation techniques and morphology on IMSI outcomes.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Intracytoplasmic morphologically selected sperm injection outcomes: the role of sperm preparation techniques

Borges

Setti

Vingris

et al. 2013

J Assist Reprod Genet

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show abstract

“…Although several studies have compared the effects of DGC and swim-up or both on sperm DNA integrity, the findings have been discordant [20,24]. Chiamchanya et al [25] compared the sperm quality Figure 2.…”

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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“…Previous publications 25,26 have shown that density gradient centrifugation increases the isolation of spermatozoa with normal morphology and DNA integrity. However, our study demonstrated that the incidence of sperm aneuploidy and DNA fragmentation was still high in males with a sperm concentration of ,5310 6 million ml 21 after the process of density gradient preparation.…”

Section: Discussionmentioning

confidence: 95%

No difference in high-magnification morphology and hyaluronic acid binding in the selection of euploid spermatozoa with intact DNA

Mongkolchaipak¹,

Vutyavanich²

2013

Asian J Androl

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In this study, we compared conventional sperm selection with high-magnification morphology based on the motile sperm organellar morphology examination (MSOME) criteria, and hyaluronic acid (HA) binding for sperm chromosome aneuploidy and DNA fragmentation rates. Semen from 50 severe male factor cases was processed through density gradient centrifugation, and subjected to sperm selection by using the conventional method (control), high magnification at 36650 or HA binding. Aneuploidy was detected by fluorescence in situ hybridization with probes for chromosomes 13, 18, 21, X and Y, and DNA fragmentation by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) method. Spermatozoa selected under high-magnification had a lower DNA fragmentation rate (2.6% vs. 1.7%; P50.032), with no significant difference in aneuploidy rate (0.8% vs 0.7%; P50.583), than those selected by the HA binding method. Spermatozoa selected by both methods had much lower aneuploidy and DNA fragmentation rate than the controls (7% aneuploidy and 26.8% DNA fragmentation rates, respectively). In the high-magnification group, the aneuploidy rate was lower when the best spermatozoa were selected than when only the second-best spermatozoa were available for selection, but the DNA fragmentation rate was not different. In conclusion, sperm selection under high magnification was more effective than under HA binding in selecting spermatozoa with low DNA fragmentation rate, but the small difference (0.9%) might not be clinically meaningful. Both methods were better than the conventional method of sperm selection. Keywords: aneuploidy; DNA fragmentation; hyaluronic acid binding; intracytoplasmic sperm injection (ICSI); intracytoplasmic morphologically selected sperm injection (IMSI); motile sperm organellar morphology examination; physiologic intracytoplasmic sperm injection INTRODUCTION Intracytoplasmic sperm injection (ICSI) is a very effective method for the treatment of severe male factor infertility. It allows the use of a single motile spermatozoon to fertilise an oocyte. 1 The technique is so effective that fertilization of an oocyte can be achieved even with the use of a spermatozoon with severe DNA fragmentation. 2 Men with severe male factor infertility are known to have a higher frequency of sperm aneuploidies and DNA fragmentation than fertile men. [3][4][5][6][7][8] Although the consequences of inseminating oocytes with abnormal spermatozoa are not known for certain, there is increasing evidence that such circumstances may cause poor fertilization, defective pre-implantation embryonic development, and high rates of miscarriage and morbidity in the offspring, including childhood cancer. 8 One major challenge ICSI facing is, therefore, the selection of the best spermatozoon for micro-insemination.Many studies have shown that intracytoplasmic morphologicallyselected sperm injection (IMSI) can significantly increase the fertilization and pregnancy rate of ICSI, while decreasing its abortion rate. [9][10][11] The disadvantage...

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The use of two density gradient centrifugation techniques and the swim-up method to separate spermatozoa with chromatin and nuclear DNA anomalies

Cited by 214 publications

References 23 publications

Intracytoplasmic morphologically selected sperm injection outcomes: the role of sperm preparation techniques

Intracytoplasmic morphologically selected sperm injection outcomes: the role of sperm preparation techniques

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

No difference in high-magnification morphology and hyaluronic acid binding in the selection of euploid spermatozoa with intact DNA

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