Washing and Concentration of Human Semen by Percoll Density Gradients and Its Application to Aih

Iizuka, R; Kaneko, Satoru; Kobanawa, K.; Kobayashi, Toshifumi

doi:10.3109/01485018808987061

Cited by 23 publications

(8 citation statements)

References 9 publications

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“…Percoll TM gradient density (Kabi Pharmacia, Uppsala, Sweden) has been used effectively, and has been approved for sperm selection since 1988 [8][9][10][11] owing to its superior selection ability compared with simple washing or swim-up preparations [12][13][14][15]. However, several preparation media that use centrifugation gradients have been studied [8][9][10][11], including Isolate TM , SpermGrad TM , Sil-Select Plus TM , Optiprep TM and PureSperm  . Among them, PureSperm yielded similar sperm parameters to those of Percoll and could be considered a suitable substitute for Percoll, which was abandoned in 1996 owing to toxicity [16].…”

Section: Resultsmentioning

confidence: 99%

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

Chiamchanya¹,

Kaewnoonual²,

Visutakul³

et al. 2009

Asian J Androl

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Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media, such as PureSperm  (Nidacon, Gothenburg, Sweden), Sil-Select Plus TM (Fertipro, Beernem, Belgium) and SpermGrad TM (Vitrolife, Gothenburg, Sweden). The differences in mean percentages of semen parameters were assessed by repeated measures analysis. Correlations of sperm DNA damage, as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and of protamine deficiency, as measured by chromomycin A3 (CMA3) staining with sperm parameters, were determined by Pearson's correlation. After preparation with all three media, sperm concentrations decreased (P < 0.05) while percentages of sperm with normal morphology increased (P < 0.05). Percentages of sperm motility, rapid motility and progressive motile concentration (PMC) increased (P < 0.05) for each of these parameters, PureSperm preparation gave the best results (P < 0.05). The percentage of DNA damage decreased in the PureSperm and Sil-Select Plus preparations (17.9% and 31.3%, respectively, P < 0.05) and increased in the SpermGrad preparation (56.3%, P < 0.05). Protamine deficiency also decreased in all three kinds of media, 59.3%, 47.7% and 40.3% for PureSperm, Sil-Select Plus and SpermGrad preparations, respectively (P < 0.05). The percentage of DNA-damaged sperm was negatively correlated with the percentages of sperm motility, rapid motility and PMC, but was positively correlated with static motility (P < 0.05). This comparative study and correlation analysis revealed that PureSperm preparation yielded sperm with the best motility and the lowest percentage of protamine deficiency. The Sil-Select Plus preparation yielded sperm with the lowest amount of DNA damage. The SpermGrad preparation had a high percentage of sperm with normal morphology, but also had the highest percentage of sperm with DNA damage. Sperm DNA damage was correlated with percentages of sperm motility, rapid motility, static motility and PMC. IntroductionIt is well known that semen quality is a significant factor for determining the success of pregnancy, apart from oocyte and endometrial quality. A semen prepa- 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 Comparative study of the effects of three semen preparation media Charoenchai Chiamchanya et al.

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Section: Resultsmentioning

confidence: 99%

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

Chiamchanya¹,

Kaewnoonual²,

Visutakul³

et al. 2009

Asian J Androl

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“…A Percoll separation step was added to the sperm preparation protocol, and this treatment resulted in zona binding and acrosome reaction of a small number of control sperm. Percoll gradients have been utilized for human sperm preparation to enhance the percentage of motile sperm and to remove seminal plasma (Iizuka et al, 1988;Pardo and Bancells, 1989).…”

Section: Discussionmentioning

confidence: 99%

Separate effects of caffeine and dbcAMP on macaque sperm motility and interaction with the zona pellucida

VandeVoort

Tollner

Overstreet

1994

Molecular Reproduction Devel

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Capacitation of macaque sperm with caffeine and dbcAMP is required for fertilization in vitro. This study determined the separate effects of caffeine and dbcAMP on sperm-zona pellucida binding and the acrosome reaction of zona bound sperm. Semen from 6 cynomolgus macaques was washed through 60% Percoll, resuspended, and washed with BWW media and incubated for 2.5 hr. Caffeine, dbcAMP (2 mM each), or both (1 mM each) were added to aliquots of the sperm suspensions. Immature macaque oocytes were placed into drops of sperm suspensions, coincubated with sperm for 30 sec, and either fixed immediately or removed to sperm-free media and incubated 1 hr before fixation. There were no significant differences between groups in the percentage of live, acrosome-reacted sperm in suspension. Treatment with caffeine and dbcAMP or with caffeine alone, significantly increased the number of sperm bound to each zona pellucida (96 +/- 16 and 81 +/- 17, respectively) compared to control and dbcAMP treatment (15 +/- 4 and 28 +/- 13). However, treatment with dbcAMP, alone and with caffeine, resulted in a higher percentage of acrosome-reacted sperm on the zona (15.2 +/- 2.1 and 9.0 +/- 0.6) than control or caffeine treatment (3.0 +/- 1.4 and 2.4 +/- 0.5). Effects on sperm motility consistent with hyperactivation were detected only when both caffeine and dbcAMP were present. Although both caffeine and dbcAMP are presumed to increase or to produce the same effect as increased intracellular cAMP levels, these compounds have different effects on the ability of sperm to bind to the zona and to undergo the acrosome reaction.

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“…Percoll gradient density (Kabi Pharmacia, Uppsala, Sweden) has been used extensively for spermatozoa selection. Several studies have proved Percoll's effectiveness (Iizuka et al, 1988; Mathieu et al, 1988; Punjabi et al, 1990; Claassens et al, 1996) and its comparatively better selection compared with simple washing or swim‐up preparation (Berger et al, 1985; McClure et al, 1989; Bongso et al, 1993; Moohan and Lindsay, 1995). Good clinical pregnancy rates were obtained after Percoll migration (Pardo et al, 1988; Sato et al, 1990), higher compared with swim‐up preparation (Guérin et al, 1989).…”

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confidence: 99%

Comparison of Six Density Gradient Media for Selection of Cryopreserved Donor Spermatozoa

Mousset-Siméon¹,

Rives

Masse

et al. 2004

Journal of Andrology

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The aim of our study was to evaluate the efficiency of 4 density gradient media for motile cryopreserved spermatozoa selection to Percoll (Kabi Pharmacia, Uppsala, Sweden) and to Puresperm (J.C.D. International Laboratory, L'Aigle, France). Puresperm was the new medium chosen in our laboratory in 1996 as the substitute for Percoll. The solutions tested were 3 colloidal silane-coated silica particle media (Isolate, SpermGrad-100, Sil-Select Plus) and iodixanol (Optiprep). Semen parameters analyzed after selection were concentration, motility, and morphology. Semen parameters after Puresperm gradient had similar values compared to Percoll. Optiprep was less efficient with a poor concentration. Isolate had a comparatively better concentration, but the capacity of selection was not satisfactory. SpermGrad-100 and Sil-Select Plus were less effective than Puresperm. In conclusion, Puresperm could be considered a better alternative to Percoll for cryopreserved spermatozoa migration.

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Washing and Concentration of Human Semen by Percoll Density Gradients and Its Application to Aih

Cited by 23 publications

References 9 publications

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

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

Separate effects of caffeine and dbcAMP on macaque sperm motility and interaction with the zona pellucida

Comparison of Six Density Gradient Media for Selection of Cryopreserved Donor Spermatozoa

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