A novel, biomarker-based assay to screen for dysfunctional spermatozoa

Vairo, L.; Neri, Q.V.; Rosenwaks, Zev; Travis, Alexander J.; Palermo, G.D.

doi:10.1016/j.fertnstert.2013.07.1284

Cited by 6 publications

(5 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The sperm were resuspended in mHTF with (Cap) and without (NonCap) 2‐hydroxypropyl‐β‐cyclodextrin (CD; Sigma, St. Louis, MO, catalogue # C0926) to promote capacitation. Previous studies have shown that stimulation with 2‐hydroxypropyl‐β‐cyclodextrin was as effective in promoting capacitation in human sperm at 3 hr of incubation as albumin was at 6 hr (Vairo et al, ). Following incubation, the samples were lightly fixed based on the original findings of Selvaraj et al (), and different treatments were created based on the time of their incubation and time maintained in the fix prior to analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, ejaculates were liquefied for up to 2 hr. Following liquefaction, the sperm were removed from the seminal plasma by centrifugation (Vairo et al, 2013). Following incubation, the samples were lightly fixed based on the original findings of Selvaraj et al (2006), and different treatments were created based on the time of their incubation and time maintained in the fix prior to analysis.…”

Section: Standard Sample Processingmentioning

confidence: 99%

See 1 more Smart Citation

Timing of sperm capacitation varies reproducibly among men

Ostermeier¹,

Cardona²,

Moody³

et al. 2018

Molecular Reproduction Devel

View full text Add to dashboard Cite

Sperm must mature functionally in the process of capacitation to become able to fertilize. Capacitation depends on membrane lipid changes, and can be quantitatively assessed by redistribution of the ganglioside GM1, the basis of the Cap‐Score™ sperm function test. Here, differences in Cap‐Score were compared among and within men at two time points. Ejaculates were liquefied, washed, and incubated for 3 hr under capacitating (Cap) conditions, then fixed and analyzed immediately (Day0); after being incubated 3 hr under Cap conditions then maintained 22–24 hr in fix (Day1‐fix); or after 22–24 hr incubation under Cap conditions prior to fixation (Day1). In all cases, a light fixative previously shown to allow membrane lipid movements was used. Day1‐fix and Day1 Cap‐Scores were greater than Day0 (p < 0.001; n = 25), whereas Day1‐fix and Day1 Cap‐Scores were equivalent (p = 0.43; n = 25). In 123 samples from 52 fertile men, Cap‐Score increased more than 1SD (7.7; calculated previously from a fertile cohort) from Day0 to Day1‐fix in 44% (54/123) of the samples. To test whether timing of capacitation was consistent within an individual, 52 samples from 11 fertile men were classified into either “early” or “late” capacitation groups. The average capacitation group concordance within a donor was 81%. Median absolute deviation (MAD; in Cap‐Score units) was used to assess the tightness of clustering of the difference from Day0 to Day1‐fix within individuals. The average (2.21) and median (1.98) MAD confirmed consistency within individuals. Together, these data show that the timing of capacitation differed among men and was consistent within men.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Standard Sample Processingmentioning

confidence: 99%

Timing of sperm capacitation varies reproducibly among men

Ostermeier¹,

Cardona²,

Moody³

et al. 2018

Molecular Reproduction Devel

View full text Add to dashboard Cite

show abstract

“…The capacitation stimuli consisted of 2‐hydroxypropyl‐ β‐cyclodextrin (Sigma, St. Louis, MO; reference C0926) (Osheroff et al, ; Parinaud, Vieitez, Vieu, Collet, & Perret, ). Pilot studies showed that this stimulus was as effective in promoting capacitation in human sperm, as measured with the Cap‐Score, at 3 h of incubation as albumin was at 6 h (Vairo et al, ). Following incubation, the samples were fixed with paraformaldehyde (Electron Microscopy Services: Hatfield, PA reference 15712) as described (Selvaraj et al, ).…”

Section: Methodsmentioning

confidence: 99%

Localization patterns of the ganglioside G_M1in human sperm are indicative of male fertility and independent of traditional semen measures

Cardona¹,

Neri

Simpson³

et al. 2017

Mol Reprod Dev

View full text Add to dashboard Cite

Semen analysis lacks a functional component and best identifies extreme cases of infertility. The ganglioside GM1 is known to have functional roles during capacitation and acrosome exocytosis. Here, we assessed whether GM1 localization patterns (Cap‐Score™) correspond with male fertility in different settings: Study 1 involved couples pursuing assisted reproduction in a tertiary care fertility clinic, while Study 2 involved men with known fertility versus those questioning their fertility at a local urology center. In Study 1, we examined various thresholds versus clinical history for 42 patients; 13 had Cap‐Scores ≥39.5%, with 12 of these (92.3%) achieving clinical pregnancy by natural conception or ≤3 intrauterine insemination cycles. Of the 29 patients scoring <39.5%, only six (20.7%) attained clinical pregnancy by natural conception or ≤3 intrauterine insemination cycles. In Study 2, Cap‐Scores were obtained from 76 fertile men (Cohort 1, pregnant partner or recent father) and compared to 122 men seeking fertility assessment (Cohort 2). Cap‐Score values were normally distributed in Cohort 1, with 13.2% having Cap‐Scores more than one standard deviation below the mean (35.3 ± 7.7%). Significantly, more men in Cohort 2 had Cap‐Scores greater than one standard deviation below the normal mean (33.6%; p = 0.001). Minimal/no relationship was found between Cap‐Score and sperm concentration, morphology, or motility. Together, these data demonstrate that Cap‐Score provides novel, clinically relevant insights into sperm function and male fertility that complement traditional semen analysis. Furthermore, the data provide normal reference ranges for fertile men that can help clinicians counsel couples toward the most appropriate fertility treatment.

show abstract

“…Ejaculates within a sealed specimen container (Fisher Scientific, 14-375-462) were liquefied at 37°C in an air incubator, and then removed from the seminal plasma by centrifugation through (Osheroff et al, 1999;Parinaud, Vieitez, Vieu, Collet, & Perret, 2000). Pilot studies showed that this stimulus was as effective in promoting capacitation in human sperm, as measured by G M1 localization patterns, at 3 hr of incubation as albumin was at 6 hr (Vairo et al, 2013). Following incubation, all samples were fixed with paraformaldehyde (Electron Microscopy Sciences; Hatfield, PA), as previously described (Selvaraj et al, 2006).…”

Section: Standard Sample Processingmentioning

confidence: 99%

Validation of a laboratory-developed test of human sperm capacitation

Moody¹,

Cardona²,

Simpson³

et al. 2017

Mol Reprod Dev

View full text Add to dashboard Cite

Sperm must undergo capacitation to become fertilization competent. Here we validated that monosialotetrahexosylganglioside (GM1) localization patterns, which were assessed in the Cap‐Score™ Sperm Function Test, reflect a capacitated state in human sperm. First, we defined patterns representing sperm that do or do not respond to stimuli for capacitation. Sperm with “capacitated” patterns had exposed acrosomal carbohydrates and underwent acrosome exocytosis in response to calcium ionophore (A23187). Precision was evaluated by percent change of the Cap‐Score measured for 50, 100, 150, and 200 sperm. Changes of 11%, 6%, and 5% were observed (n ≥ 23); therefore, we counted ≥150 sperm per condition. Variance within and between readers was evaluated using 20 stitched image files generated from unique ejaculates. Two trained readers randomly resampled each image 20 times, reporting an average standard deviation of 3 Cap‐Score units and coefficient of variation of 13% when rescoring samples, with no difference between readers. Semen liquefaction times ≤2 hr and mechanical liquefaction with Pasteur or wide‐orifice transfer pipettes did not alter Cap‐Score values. However, liquefaction with chymotrypsin (p = 0.002) and bromelain (p = 0.049) reduced response to capacitating stimuli and induced membrane damage, while counterintuitively improving sperm motility. Together, these data validate the Cap‐Score assay for the intended purpose of providing information on sperm capacitation and male fertility. In addition to its clinical utility as a diagnostic tool, this test of sperm function can reveal the impact of common practices of semen handling on the ability of sperm to respond to capacitation stimuli.

show abstract

A novel, biomarker-based assay to screen for dysfunctional spermatozoa

Cited by 6 publications

References 0 publications

Timing of sperm capacitation varies reproducibly among men

Timing of sperm capacitation varies reproducibly among men

Localization patterns of the ganglioside G_M1in human sperm are indicative of male fertility and independent of traditional semen measures

Validation of a laboratory-developed test of human sperm capacitation

Contact Info

Product

Resources

About

A novel, biomarker-based assay to screen for dysfunctional spermatozoa

Cited by 6 publications

References 0 publications

Timing of sperm capacitation varies reproducibly among men

Timing of sperm capacitation varies reproducibly among men

Localization patterns of the ganglioside GM1in human sperm are indicative of male fertility and independent of traditional semen measures

Validation of a laboratory-developed test of human sperm capacitation

Contact Info

Product

Resources

About

Localization patterns of the ganglioside G_M1in human sperm are indicative of male fertility and independent of traditional semen measures