Properties of sperm separated using Percoll and IxaPrep density gradients. A comparison made using CASA, longevity, morphology and the acrosome reaction

McCann, Christine; Chantler, Eric

doi:10.1046/j.1365-2605.2000.00228.x

Cited by 27 publications

(19 citation statements)

References 21 publications

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“…This sperm separation technique could effectively acquire sperm populations with high motility and normal morphology free of seminal plasma, debris, microbial contamination, and residual cells (immature germ cells or polynuclear cells). This was confirmed by our microscopy inspection and has been shown in previous reports [16,17,19,21,22]. Sperm motility was monitored Fig.…”

Section: Discussionsupporting

confidence: 90%

Presence, localization, and origin of clusterin in normal human spermatozoa

Han

Wang

Cheng

et al. 2012

J Assist Reprod Genet

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Purpose Clusterin in mammalian semen is a secretory form of clusterin (sCLU) with the heterodimeric structure. It is secreted by the epididymis and seminal vesicle. It is generally agreed that clusterin mainly exists on the surface of abnormal spermatozoa and is implicated in decreased sperm motility, sperm aggregation and infertility. However, few studies observe clusterin in normal spermatozoa, which is presumed to be a novel form. Up to now, the systematical information about the presence, localization, origin and function of clusterin in normal human spermatozoa has yet not been established. The aim of our current study is to systematically research clusterin in normal human spermatozoa.Methods We detected the presence of clusterin via western blot, explored the localization of clusterin using immunofluorescence, and investigated the origin and distribution of clusterin in human testis by western blot and immunohistochemistry. Results We found native clusterin in the inner plasma membrane of normal human spermatozoa. It was derived from the testis and showed similar molecular weight and heterodimeric structure compared with sCLU in semen and on the surface of abnormal spermatozoa. Conclusion Clusterin in normal spermatozoa should be self-synthesized during the later stage of spermatogenesis. The different localization and origin suggested that the clusterin observed by us may be a novel form compared with conventional sCLU on the surface of abnormal spermatozoa.

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Section: Discussionsupporting

confidence: 90%

Presence, localization, and origin of clusterin in normal human spermatozoa

Han

Wang

Cheng

et al. 2012

J Assist Reprod Genet

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“…However, toxic effects of the Percoll † gradient on sperm and embryos have been described (De Vos et al 1997); these were attributed to polyvinylpyrrolidone (PVP). Therefore, although the use of Percoll † in human-assisted reproduction is prohibited worldwide (McCann and Chantler 2000), and its use in animals is being challenged, it is still a method of choice for animal assisted reproductive techniques. Hence, alternative protocols are needed.…”

Section: And Schill 2003) a Percollmentioning

confidence: 99%

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

et al. 2014

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. 2014. Isolate † and Optiprep † minigradients as alternatives for sperm selection in bovine in vitro embryo production. Can. J. Anim. Sci. 94: 35Á42. The objective of this study was to evaluate alternatives in small volumes to conventional gradient of Percoll † on semen quality, in vitro embryo production, sex ratio and embryo survival after vitrification. Thawed semen was randomly allocated to one of four density gradient selection methods: (1) conventional Percoll † (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), and (4) MiniOptiprep (MO). Sperm kinetics and quality were evaluated. Use of P, MP and MI gradients did not affect sperm motility (P!0.05). However, there was a decrease in total and progressive sperm motility in MO (70.8 and 51.3% vs. 87.3 and 69.5% for P; 87.3 and 73% for MP; 92.3 and 78.8% for MI; PB0.05). The MO had lower membrane integrity compared with P, MP and MI (39.7 vs. 70.5, 72.3, 63.8%, respectively, PB0.05). The percentage of blastocysts produced was higher in MI than in MP and MO (21.1 vs. 16.1 and 16.9%, PB0.05) and similar to P (18.4%; P!0.05). Sex ratio and embryo survival after vitrification were similar among groups (P!0.05). Semen selected by Isolate and Optiprep gradient, at the concentrations and small volumes used, demonstrated similar characteristics and in vitro embryo production to conventional Percoll † gradient. † , en petits volumes, sur la qualite´de la semence, la production in vitro d'embryons, le rapport des sexes et la survie de l'embryon apre`s vitrification. Des semences de´congele´es ont e´te´attribue´es ale´atoirement a`une de quatre me´thodes de se´lection par gradient de densite´: (1) Percoll † traditionnel (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), et (4) MiniOptiprep (MO). La cine´tique et la qualite´des spermatozoı¨des ont e´te´e´value´es. L'utilisation des gradients P, MP et MI n'ont eu aucun effet sur la motilite´des spermatozoı¨des (P!0,05). Il y avait, cependant, une diminution des motilite´s totales et progressives dans le gradient MO (70,8 et 51,3 % c. 87,3 et 69,5 % pour P; 87,3 et 73 % pour MP; 92,3 et 78,8 % pour MI; PB0,05). Le gradient MO avait une inte´grite´de membrane infe´rieure aux gradients P, MP et MI (39,7 c. 70,5; 72,3; 63,8 % respectivement, PB0,05). Le pourcentage de blastocystes produits e´tait plus e´leve´apre`s gradient MI qu'apre`s les gradients MP et MO (21,1 c. 16,1 et 16,9 %, P B0,05) et similaire apre`s gradient P (18,4 %; P!0,05). Le rapport des sexes et la survie de l'embryon apre`s vitrification e´taient similaires parmi tous les groupes (P !0,05). Les semences choisies par gradients Isolate et Optiprep, aux concentrations et aux petits volumes utilise´s, de´montrent des caracte´ristiques et une production in vitro d'embryons similaires a`celles choisies par gradients Percoll † traditionnels.

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“…Although in most mammalian species the basal level of SDF after thawing does not increase when compared with that observed before freezing, the rSDF after thawing tends to be higher [López-Fernández et al 2007;. In general, sperm motility, viability, and most motion characteristics are significantly affected in samples prepared by 'swim-up' [Esteves et al 2000] or gradientisolation techniques [McCann and Chantler 2002;Edelstein et al 2008]. For example, swim-up isolates sperm with a greater ability to undergo hypo-osmotic swelling, but does not change the rate of spontaneous acrosome reaction, even though in vitro capacitation of spermatozoa selected by swim-up enhances the acrosome reaction rate [Esteves et al 2000;Zaneveld et al 1991].…”

Section: Introductionmentioning

confidence: 99%

DNA Fragmentation Dynamics in Fresh Versus Frozen Thawed Plus Gradient-Isolated Human Spermatozoa

Gosálvez

Torre

López‐Fernández

et al. 2010

Systems Biology in Reproductive Medicine

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The aim of this study was to compare the rate of sperm DNA fragmentation (rSDF; increase of SDF over time) in fresh and gradient isolated frozen-thawed semen samples from male sperm donors of proven fertility. SDF was assessed in the two samples obtained from the same fifteen male donors after 0.5, 1.5, 4.5, 6, 24, 48, and 72 h of incubation in a humidified atmosphere of 5% CO 2 in air at 371C. Analysis was performed based on chromatin dispersion patterns evaluated using the Halosperms kit. No significant differences in SDF were obtained when fresh and gradient-isolated frozen-thawed spermatozoa were compared at baseline. However, the rSDF shown by the two samples differed and gradient-isolation selected for sperm subpopulations showing a lower variance for SDF. At the individual level, sperm selection by density gradient purification in frozen-thawed samples did not improve the levels of SDF when compared with the values obtained in fresh samples at baseline.

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Properties of sperm separated using Percoll and IxaPrep density gradients. A comparison made using CASA, longevity, morphology and the acrosome reaction

Cited by 27 publications

References 21 publications

Presence, localization, and origin of clusterin in normal human spermatozoa

Presence, localization, and origin of clusterin in normal human spermatozoa

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

DNA Fragmentation Dynamics in Fresh Versus Frozen Thawed Plus Gradient-Isolated Human Spermatozoa

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Properties of sperm separated using Percoll and IxaPrep density gradients. A comparison made using CASA, longevity, morphology and the acrosome reaction

Cited by 27 publications

References 21 publications

Presence, localization, and origin of clusterin in normal human spermatozoa

Presence, localization, and origin of clusterin in normal human spermatozoa

Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production

DNA Fragmentation Dynamics in Fresh Versus Frozen Thawed Plus Gradient-Isolated Human Spermatozoa

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Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production