Improved motile sperm recovery by a hyperosmotic percoll gradient

Carbone, Dominick J.; Seifarth, Karen; Thomas, Anthony J.; Agarwal, Ashok

doi:10.1007/bf02766147

Cited by 4 publications

(2 citation statements)

References 20 publications

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“…Sperm incubation in media with high osmolarity lead to a progressive reduction of sperm total motility percentages and kinetic characteristics so that when sperm were suspended in medium with an osmolarity close to 600 mOsm residual sperm total motility was as high as 4.4%. The importance of medium osmolarity in the regulation of sperm motility has also been demonstrated previously during human sperm isolation using hyperosmolar Percoll preparations (Carbone et al, 1997). Finally, the demonstration that sperm motility characteristics improve when washed from seminal plasma with high osmolarity and resuspended in medium with lower osmolarity further strengthens the role of osmolarity in sperm motility regulation.…”

Section: Discussionsupporting

confidence: 70%

Role of seminal osmolarity in the regulation of human sperm motility

et al. 2002

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Mammalian sperm at ejaculation are suspended in the seminal plasma, a heterogeneous mixture deriving from the testicular/epididymal fluid and from secretions of seminal vesicles, prostate and bulbourethral glands. Biochemical characteristics of seminal fluid change along the male reproductive tract when considering its inorganic and organic composition and pH but it is known that in each region of the male genital tract seminal osmolarity is higher than that of serum. It has been previously demonstrated that in invertebrate and vertebrate sperm, seminal plasma osmolarity influences sperm motility and activity, and human sperm have been shown to possess osmosensitive calcium entry pathway that controls important functions such as acrosome reaction and oocyte penetration. In the present study, we have determined seminal plasma osmolarity in a large number of normozoospermic fertile and asthenozoospermic infertile subjects correlating it with sperm motility percentages and kinematic characteristics determined utilizing a computerized motion analysis system. Our results confirm that seminal plasma osmolarity is higher than that of serum (336.1 +/- 20.2 vs. 291.1 +/- 6.9 mOsm/L, respectively). Normozoospermic subjects show seminal osmolarity values that are significantly lower with respect to asthenozoospermic patients (317.8 +/- 12.2 vs. 345.2 +/- 22.6 mOsm/L, p<0.001), irrespective of the cause of asthenozoospermia. Seminal plasma osmolarity negatively correlates with sperm progressive motility percentages and kinetic characteristics (curvilinear velocity, linear velocity, linear coefficient and lateral displacements of sperm head). Furthermore, when sperm from fertile subjects were suspended in medium with an osmolarity increasing from 300 to 600 mOsm, sperm motility percentages and kinetics characteristics were progressively reduced and nearly abolished when medium osmolarity was 600 mOsm. On the contrary, when sperm from asthenozoospermic subjects with high semen osmolarity were resuspended in medium with lower osmolarity, sperm motility parameters improved significantly. Sperm motility parameters did not correlate with seminal plasma concentrations of sodium, potassium, chloride with a weak correlation only with seminal calcium concentration. No correlations are present between seminal plasma osmolarity and ionic composition. In conclusion, the present study confirms and extends the knowledge that, in human, seminal plasma osmolarity is higher than that of serum and demonstrates that seminal osmolarity influences sperm motility characteristics and then it may contribute to the pathogenesis of some forms of asthenozoospermia and male infertility.

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Section: Discussionsupporting

confidence: 70%

Role of seminal osmolarity in the regulation of human sperm motility

et al. 2002

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“…Two discontinuous gradients with low centrifugal force applied were used clinically. Carbone et al [14] reported that use of a hyperosmotic gradient increased the yield of motile sperm, and Makler et al [8] reported that prolonging the centrifugation time to 60 min resulted in more nonmotile sperm being added to motile sperm. Under these conditions no attempt was made to reach a static endpoint.…”

Section: Discussionmentioning

confidence: 99%

Effect of Density Gradient Osmolality on Specific Gravity of Bull Sperm and Their Separation, but Not on the Basis of Sex.

Hough

Foote

2002

Journal of Reproduction and Development

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Abstract.The most effective quantitative separation of ejaculates of semen into subsets of sperm differing in quality by density gradients requires information on the specific effects of processing conditions on sperm density. Reported density of bull sperm varies greatly. This research was undertaken to study the effects of gradient osmolality on sperm density and on separation using gradients with low viscosity and low toxicity to sperm. Semen was available from bulls used for artificial insemination. Cell volume studies revealed that live sperm volume was altered by more than 25% by exposure to solutions differing in osmolality, resulting in predictable differences in density measurable by density gradients. Control studies indicated that discontinuous Nycodenz ® gradients differing by less than 1% in density could be prepared. Following centrifugation at 100,000 × g for 1 h fractions containing 96-99% unstained (live) and 91% motile sperm versus a fraction with 100% nonmotile sperm were obtained. The densities (g/ml) of live sperm separated on low osmolality gradients (220-237 mOsm), and on high osmolality gradients (342-345 mOsm) were 1.125 versus 1.145, respectively. Killed sperm on both gradients had a density of 1.233 g/ml. These gradients permit quantitative separation of sperm varying in membrane integrity and ability to function under different conditions, but do not permit sexing.

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Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation

Claassens¹,

Menkveld²,

Harrison³

1998

Human Reproduction

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Silane-coated silica particle solutions (ISolate(TM) and PureSperm)TM)) and iodixanol (OptiPrep(TM)) were compared to polyvinylpyrrolidone (PVP)-coated silica particles (Percoll(TM)) in their efficacy to recover spermatozoa by gradient centrifugation for use in assisted reproductive procedures. Efficacy was assessed in terms of percentages of sperm recovery, sperm vitality and motility, normal sperm morphology and normal sperm chromatin condensation. No significant difference was found in the recovery of spermatozoa for men with both normal sperm counts and oligozoospermia, between PVP-coated and silane-coated particle solutions. Iodixanol had significantly lower sperm recovery compared to the other products. Sperm vitality, progressive motility, normal morphology and normal chromatin condensation did not differ significantly between any of the sperm isolation products.

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Improved motile sperm recovery by a hyperosmotic percoll gradient

Abstract: ~TRODUCTIONThe ideal semen processing technique should result in the selective recovery of good-quality motile sperm while maintaining a high total yield of recovered sperm. Several procedures for retrieving motile sperm from the ejaculate have been described, including

Cited by 4 publications

References 20 publications

Role of seminal osmolarity in the regulation of human sperm motility

Role of seminal osmolarity in the regulation of human sperm motility

Effect of Density Gradient Osmolality on Specific Gravity of Bull Sperm and Their Separation, but Not on the Basis of Sex.

Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation

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