Spermatozoa Recovered by Ixaprep Gradient have Improved Longevity and Better Motion Characteristics than Those by Percoll Gradient

Yang, Jyh‐Chin; Wu, Ming‐Jung; Chen, C.-D.; Chao, Kuang Han; Chen, S.-U.; Ho, Hong Nerng; Yang, Yu Shih

doi:10.3109/01485019808987947

Cited by 5 publications

(9 citation statements)

References 23 publications

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“…Human spermatozoa are prone to ROS‐induced damage due to a higher level of polyunsaturated fatty acids in their membranes compared with somatic cells. The higher level of immotile and abnormal spermatozoa that we have found in IxaPrep‐separated spermatozoa could indicate a greater potential to generate ROS, though Yang et al . (1998 ) considered that ROS‐generating systems were possibly removed by IxaPrep separation.…”

Section: Discussionsupporting

confidence: 93%

“…IxaPrep has been proposed as a substitute for Percoll for processing spermatozoa in human ART ( Yang et al ., 1998 ). We have compared the separation and the properties of spermatozoa from semen processed using both Percoll and IxaPrep.…”

Section: Discussionmentioning

confidence: 99%

“…IxaPrep does not contain PVP, demonstrates a low toxicity and has the lowest endotoxin level of any available density gradient material (0.5 EU/mL). Yang et al . (1998 ) found that IxaPrep provided a comparable separation of spermatozoa to Percoll.…”

Section: Introductionmentioning

confidence: 99%

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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

McCann

Chantler

2000

Int J of Andrology

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Since the withdrawal of Percoll from use in human assisted reproduction techniques in 1996, alternative density gradients have been sought. IxaPrep is one candidate that has low toxicity and can separate spermatozoa. Fractions of spermatozoa from normozoospermic men, separated by Percoll and IxaPrep gradient centrifugation, were assessed for sperm recovery, motility, morphology and the ability to undergo the acrosome reaction. The Percoll fractions had a significantly higher sperm recovery, motile count, rapid motile count, VAP, VSL, and VCL than those obtained using IxaPrep, both immediately after separation and up to 24 h later. In contrast, ALH, level of abnormal sperm morphology and the ability to acrosome react were not statistically different in sperm fractions separated using either gradient forming material.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

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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

McCann

Chantler

2000

Int J of Andrology

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“…While Claassens et al [61] and Söderlund and Lundin [62] did not find differences in the recovery rate between Percoll ® and PureSperm ® , Chen and Bongso [63], depending on the number of layers included for the density gradient, reported significantly higher values for PureSperm ® . For IxaPrep ® it is even more confusing because Yang et al [64] found no difference to Percoll ® , while Makkar et al [14] found the replacement substance more effective. On the other hand, McCann and Chantler [65] as well as Ding et al [66] found Percoll ® superior.…”

Section: Sperm Separation Techniquesmentioning

confidence: 99%

Sperm preparation for ART

Henkel

Schill

2003

Reprod Biol Endocrinol

423

175

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The onset of clinical assisted reproduction, a quarter of a century ago, required the isolation of motile spermatozoa. As the indication of assisted reproduction shifted from mere gynaecological indications to andrological indications during the years, this urged andrological research to understand the physiology of male germ cell better and develop more sophisticated techniques to separate functional spermatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. Initially, starting from simple washing of spermatozoa, separation techniques, based on different principles like migration, filtration or density gradient centrifugation evolved. The most simple and cheapest is the conventional swim-up procedure. A more sophisticated and most gentle migration method is migration-sedimentation. However, its yield is relatively small and the technique is therefore normally only limited to ejaculates with a high number of motile spermatozoa. Recently, however, the method was also successfully used to isolate spermatozoa for intracytoplasmic sperm injection (ICSI). Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient centrifugation with different media. Since Percoll® as a density medium was removed from the market in 1996 for clinical use in the human because of its risk of contamination with endotoxins, other media like IxaPrep®, Nycodenz, SilSelect®, PureSperm® or Isolate® were developed in order to replace Percoll®. Today, an array of different methods is available and the selection depends on the quality of the ejaculates, which also includes production of reactive oxygen species (ROS) by spermatozoa and leukocytes. Ejaculates with ROS production should not be separated by means of conventional swim-up, as this can severely damage the spermatozoa. In order to protect the male germ cells from the influence of ROS and to stimulate their motility to increase the yield, a number of substances can be added to the ejaculate or the separation medium. Caffeine, pentoxifylline and 2-deoxyadenosine are substances that were used to stimulate motility. Recent approaches to stimulate spermatozoa include bicarbonate, metal chelators or platelet-activating factor (PAF). While the use of PAF already resulted in pregnancies in intrauterine insemination, the suitability of the other substances for the clinical use still needs to be tested. Finally, the isolation of functional spermatozoa from highly viscous ejaculates is a special challenge and can be performed enzymatically to liquefy the ejaculate. The older method, by which the ejaculate is forcefully aspirated through a narrow-gauge needle, should be abandoned as it can severely damage spermatozoa, thus resulting in immotile sperm.

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“…A Computer-Aided Sperm Analyzer (HTM-IVOS Version 10.6 CASA system; Hamilton Thorn Research, Danvers, MA, USA) was used to assess the following motion parameters in each sample [5,23]: straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), beat cross frequency (BCF), straightness (STR), linearity (LIN), and percentage motility. The analysis conditions were 30 frames acquired; 60 Hz frame rate; minimum contrast of 80; minimum cell size of 3 pixels; straightness (STR) threshold of 80%; medium VAP cutoff of 25 mm=s; low VAP cutoff of 5 mm=s; low VSL cutoff of 11.0; nonmotile head size of 6 pixels; nonmotile head intensity of 160; static size limits of 1.0-2.90; static intensity limits of 0.60-1.40; static elongation limits of 0-80; slow motility was considered positive; 1.95 magnification; 2400 LED brightness; 37 C. At the outset of each assessment, the accuracy of the IVOS data was verified by utilizing the ''playback'' feature.…”

Section: Motility Parametersmentioning

confidence: 99%

Effects of Fertilization Promoting Peptide, Adenosine, and Pentoxifylline on Thawed Human Sperm

Huang

Chen

Chao

et al. 2003

Archives of Andrology

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Spermatozoa Recovered by Ixaprep Gradient have Improved Longevity and Better Motion Characteristics than Those by Percoll Gradient

Cited by 5 publications

References 23 publications

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

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

Sperm preparation for ART

Effects of Fertilization Promoting Peptide, Adenosine, and Pentoxifylline on Thawed Human Sperm

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