A comparison of BoviPure® and Percoll® on bull sperm separation protocols for IVF

Samardžija, Marko; Karadjole, Martina; Matković, Mario; Cergolj, Marijan; Getz, Iva; Dobranić, Tomislav; Tomašković, Antun; Petrić, Jakša; Šurina, Jadranka; Grizelj, Juraj; Karadjole, Tugomir

doi:10.1016/j.anireprosci.2005.04.005

Cited by 60 publications

(62 citation statements)

References 26 publications

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“…Sperm processing procedures used for ART are aimed at improving sperm quality by increasing the number of progressive motile and morphologically normal sperm cells (52). Sperm preparation by density-gradient centrifugation allows the selection of good-quality mature and actively motile spermatozoa, depending on their size, shape, and density (2, 5).…”

Section: Discussionmentioning

confidence: 99%

Association of sperm apoptosis and DNA ploidy with sperm chromatin quality in human spermatozoa

Mahfouz

Sharma

Said³

et al. 2009

Fertility and Sterility

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Section: Discussionmentioning

confidence: 99%

Association of sperm apoptosis and DNA ploidy with sperm chromatin quality in human spermatozoa

Mahfouz

Sharma

Said³

et al. 2009

Fertility and Sterility

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“…Despite progress in identifing the most suitable techniques, controversial results in the sperm quality parameters have been published by several authors. Therefore, there is a need to understand human and animal sperm selection methods [Cisale et al 2001;Parrish and Foote 1987;Said and Land 2011;Samardzija et al 2006b;Sharma et al 1999]. Sperm selection techniques are usually directed towards separating optimal sperm subpopulations related to sperm motility.…”

Section: Discussionmentioning

confidence: 99%

Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model

García-Herreros

Leal

2014

Systems Biology in Reproductive Medicine

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To cite this article: Manuel García-Herreros & Claudia L. V. Leal (2014) Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model, Systems Biology in Reproductive Medicine, 60:6, 338-347, DOI: 10.3109/19396368.2014 RESEARCH ARTICLEComparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model AbstractThe effect of different sperm washing-selection methods on sperm morphometric characteristics as a study to detect differences in the subpopulational structure has been carried out in detail in a bovine model. Cryopreserved sperm samples from 5 bulls were thawed, pooled, and processed by TALP-washing centrifugation method (TWCM), selective Percoll discontinuous density-gradient centrifugation method (PDGM), and self-migration swim-up separation method (SUMM). Live-dead assay (SYBR-14/ethidium homodimer-1), chlortetracycline assay (CTC), and sperm motility were assessed, and aliquots of sperm were processed for automated sperm morphometry analysis (ASMA) simultaneously before (raw thawed sperm used as control, RTS) and after different sperm washing-selection techniques. Deleterious effects of different methods were evident, particularly on sperm membrane integrity (p50.05) and capacitation status (p50.05). Moreover, each cell was measured for four primary dimensional parameters, and three shape parameters. All sperm morphometric parameters evaluated were analyzed by principal component analysis (PCA) and multivariate clustering analyses. PCA revealed two principal components for each sperm washing or separation method explaining more than the 91% of the variance. The number of subpopulations found was the same for all methods (four) except for PDGM (three). However, irrespective of the number of subpopulations defined by PCA and clustering analyses, the sperm subpopulational structure was found to be different and strongly influenced by the sperm selection procedure due to statistical differences found regarding the sperm biophysical changes induced by each method used (p50.001). It is concluded that different sperm washing-selection methods commonly used during IVF process, may lead to alterations in sperm morphometric characteristics, which might explain the different results seen after IVF, since an important influence of these methods on sperm subpopulational structure has been demonstrated.

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“…In all experiments, frozen semen from the same bull was used. Sperm preparation for IVF on BoviPure ® (Nidacon Laboratories AB, Gothenberg, Sweden) gradient was accomplished according to Samardzija et al (2006). Final concentration was adjusted to 1×10 6 spermatozoa/ml.…”

Section: Reagents and Mediamentioning

confidence: 99%

Effect of cysteamine supplementation during in vitro culture of early stage bovine embryos on blastocyst rate and quality

et al. 2012

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The aim of this study was to evaluate whether the addition of cysteamine to the in vitro culture media enhances the yield, hatching rate, total cell number and inner cell mass/total cell number ratio of bovine embryos. A total of 933 bovine oocytes collected from ovaries of 60 slaughtered donors were subjected to in vitro maturation and in vitro fertilization. Following fertilization, embryos were cultured in synthetic oviductal fluid without glucose. After 24 h embryos were transferred into synthetic oviductal fluid with 1.5 mM glucose and 0 (control), 50, 100 and 200 µM of cysteamine. After 48 h, the embryos were transferred into synthetic oviductal fluid with glucose but without cysteamine and cultured until Day 9. The number of cleaved embryos on Day 2, the total number of blastocysts on Day 7 and the number of hatched blastocysts on Day 9 were calculated. Differential staining of inner cell mass and trophectoderm cells of blastocysts were performed on Day 7 and Day 9 of in vitro culture. Supplementation of in vitro culture media with 100 µM cysteamine increased the blastocyst yield (P < 0.05) without affecting the hatching rate. Furthermore, the embryos cultured in the presence of 100 µM cysteamine had significantly higher number of inner cell mass cells (P < 0.05) and the proportion of inner cell mass cells (P < 0.05) compared with the controls. The results of the present study demonstrated that the addition of 100 µM cysteamine to the in vitro culture media improved blastocyst production rate and enhance embryo quality, which could lead to the improvement of the in vitro culture system for bovine embryos.

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A comparison of BoviPure® and Percoll® on bull sperm separation protocols for IVF

Cited by 60 publications

References 26 publications

Association of sperm apoptosis and DNA ploidy with sperm chromatin quality in human spermatozoa

Association of sperm apoptosis and DNA ploidy with sperm chromatin quality in human spermatozoa

Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model

Effect of cysteamine supplementation during in vitro culture of early stage bovine embryos on blastocyst rate and quality

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