Human Sperm Morphology Research Articles

Effects of in-vitro application of pentoxifylline on the morphology of human spermatozoa after vitrification in asthenozoospermic patients

Cryopreservation of human spermatozoa is widely used in many assisted reproduction units to preserve male fertility [1]. Vitrification is based on the ultrarapid freezing and is routinely assayed for cryopreservation in assisted reproductive technology. Mohamed [2] showed that cryopreservation significantly affects progressive motility, viability and mitochondrial membrane potential of spermatozoa. Pentoxifylline (PX) is a phosphodiesterase considered to be a sperm movement enhancer, hyperactivation agent, inhibitor of reactive oxygen species and acrosome reaction-improving agent. The aim of our study was to evaluate the effect of in-vitro application of PX on sperm parameters and ultrastructure after vitrification. A total of 30 asthenozoospermic semen samples were selected and divided into two groups after vitrification: control (without PX) and experimental (with PX). A significant decrease in sperm motility, morphology and viability was observed post vitrification, but sperm motility was increased significantly following application of PX. On the other hand, PX did not exert any significant effect on the ultrastructure of the acrosome, plasma membrane and tail of vitrified spermatozoa.

814 Assessment of human sperm morphology: Comparison of strict Kruger's criteria versus inverted microscopy motile sperm organelle morphology examination (MSOME)

814 Assessment of human sperm morphology: Comparison of strict Kruger's criteria versus inverted microscopy motile sperm organelle morphology examination (MSOME)

Another look at human sperm morphology.

Can a standardized assessment of abnormal human sperm morphology provide additional useful information by identifying men with more severe disturbances in different types of abnormalities? Definition-based categorization of sperm head, midpiece and tail defects has shown how differently these abnormalities are distributed in fertile men and other groups of men, thus providing high and low thresholds, a starting point for diagnosis or research purposes. Several recent studies have reported indisputable genetic origins for various sperm defects. A few studies have also identified associations between environmental factors and low percentages of morphologically normal spermatozoa. Nevertheless, with the exception of rare situations in which the vast majority of spermatozoa have specific, easily characterized defects, such as 'globozoospermia', little attention has been paid to the description and precise quantification of human sperm abnormalities. The lack of standardization in the phenotyping of sperm morphological defects by conventional microscopy is a limiting factor for diagnosis and for intra- or inter-observer or centre consistency in studies investigating the causal factors and possible functional consequences of the abnormalities detected. There are currently no baseline data for abnormalities of sperm morphology based on a standardized classification, in the general population, among fertile or other groups of men. This study is based on detailed sperm abnormality datasets acquired by a standardized classification method, from several groups of men, over the same 5-year period. We studied cross-sectional data from fertile men (n = 926), male partners from infertile couples (n = 1747) and testicular cancer patients (n = 239). We used a standardized classification to analyse Shorr-stained slides, taking into account all the abnormalities encountered. Most sperm defects were significantly more frequent in infertile than in fertile men, with 20-30% of infertile men having frequencies of abnormalities above the 95th percentile in fertile men for 9 out of the 15 categories of abnormalities. Interestingly, several head abnormalities were significantly more frequent in patients with testicular cancer than in infertile men, highlighting the particular impact of this condition on sperm morphogenesis. We used the 95th percentile in fertile men as the lower threshold and the 99th percentile in infertile men as an extreme upper threshold, for the classification of morphological abnormality frequencies into three levels: low, intermediate and high. The assessment of several semen samples, with or without a genetic background, for abnormal sperm morphology, based on the percentage of normal spermatozoa, a teratozoospermia index, and the detailed profile of abnormalities categorized according to the three levels proposed, has highlighted the value of detailed phenotyping for diagnosis and research purposes. The thresholds proposed for the various categories of sperm abnormality should be considered relative rather than absolute, owing to the known sampling error related to the limited number of spermatozoa assessed per sample, or when studying the general population or populations from regions other than Western Europe. The standardized assessment of abnormal sperm morphology requires time and experience. We therefore suggest that this assessment is carried out during a first andrological check-up or for epidemiological or research studies, rather than in the routine management of infertile couples for assisted reproductive technologies. The study design used for the fertile group of men was similar to that previously used for the WHO reference values, providing a rationale for considering the 95th percentile in fertile men as the level below which abnormalities may be considered to occur at a frequency representing random background variations of a normal spermiogenesis process. The crude frequencies obtained, and the three levels of abnormality frequency proposed for each standardized category of sperm defect, provide baseline data useful for diagnosis and a starting point for future studies aiming to identify associations with genetic or environmental factors. Part of this study was supported by contract BMH4-CT96-0314 from the European Union. The authors have no competing interests to declare.

Looking forward to human sperm morphology.

Looking forward to human sperm morphology.

An efficient method for automatic morphological abnormality detection from human sperm images

Background and objectiveSperm morphology analysis (SMA) is an important factor in the diagnosis of human male infertility. This study presents an automatic algorithm for sperm morphology analysis (to detect malformation) using images of human sperm cells. MethodsThe SMA method was used to detect and analyze different parts of the human sperm. First of all, SMA removes the image noises and enhances the contrast of the image to a great extent. Then it recognizes the different parts of sperm (e.g., head, tail) and analyzes the size and shape of each part. Finally, the algorithm classifies each sperm as normal or abnormal. Malformations in the head, midpiece, and tail of a sperm, can be detected by the SMA method. In contrast to other similar methods, the SMA method can work with low resolution and non-stained images. Furthermore, an image collection created for the SMA, has also been described in this study. This benchmark consists of 1457 sperm images from 235 patients, and is known as human sperm morphology analysis dataset (HSMA-DS). ResultsThe proposed algorithm was tested on HSMA-DS. The experimental results show the high ability of SMA to detect morphological deformities from sperm images. In this study, the SMA algorithm produced above 90% accuracy in sperm abnormality detection task. Another advantage of the proposed method is its low computation time (that is, less than 9s), as such, the expert can quickly decide to choose the analyzed sperm or select another one. ConclusionsAutomatic and fast analysis of human sperm morphology can be useful during intracytoplasmic sperm injection for helping embryologists to select the best sperm in real time.

The application of scanning near field optical imaging to the study of human sperm morphology.

BackgroundThe morphology of spermatozoa is a fundamental aspect to consider in fertilization, sperm pathology, assisted reproduction and contraception. Head, neck, midpiece, principal and terminal part of flagellum are the main sperm components to investigate for identifying morphological features and related anomalies. Recently, scanning near-field optical microscopy (SNOM), which belongs to the wide family of nanoscopic techniques, has opened up new routes for the investigation of biological systems. SNOM is the only technique able to provide simultaneously highly resolved topography and optical images with a resolution beyond the diffraction limit, typical of conventional optical microscopy. This offers the advantage to obtain complementary information about cell surface and cytoplasmatic structures.ResultsIn this work human spermatozoa both healthy and with morphological anomalies are analyzed by SNOM, to demonstrate the potentiality of such approach in the visualization of sperm morphological details. The combination of SNOM topography with optical (reflection and transmission) images enables to examine typical topographic features of spermatozoa together with underlying cytoplasmic structures. Indeed the head shape and inner components as acrosome and nucleus, and the organization of mitochondria in the midpiece region are observed. Analogously for principal tract of the tail, the ridges and the columns are detected in the SNOM topography, while their internal arrangement can be observed in the corresponding SNOM optical transmission images, without requiring specific staining procedures or invasive protocols.ConclusionsSuch findings demonstrate that SNOM represents a versatile and powerful tool to describe topographical and inner structural details of spermatozoa simultaneously. This analysis could be helpful for better characterizing several morphological anomalies, often related to sperm infertility, which cannot be examined by conventional techniques all together.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-014-0061-5) contains supplementary material, which is available to authorized users.

Ultrastructural Morphology of Sperm from Human Globozoospermia.

Globozoospermia is a rare disorder characterized by the presence of sperm with round head, lacking acrosome. Coiling tail around the nucleus has been reported since early human studies, but no specific significance has conferred it. By contrast, studies on animal models suggest that coiling tail around the nucleus could represent a crucial step of defective spermatogenesis, resulting in round-headed sperm. No observations, so far, support the transfer of this hypothesis to human globozoospermia. The purpose of this work was to compare ultrastructural morphology of human and mouse model globozoospermic sperm. Sperm have been investigated by using scanning and transmission electron microscopy. The images that we obtained show significant similarities to those described in GOPC knockout mice, an animal model of globozoospermia. By using this model as reference, we were able to identify the probable steps of the tail coiling process in human globozoospermia. Although we have no evidence that there is the same pathophysiology in man and knocked-out mouse, the similarities between these ultrastructural observations in human and those in the experimental model are very suggestive. This is the first demonstration of the existence of relevant morphological homologies between the tail coiling in animal model and human globozoospermia.

Assessment of human sperm morphology

Assessment of human sperm morphology

Probing the effect of human normal sperm morphology rate on cycle outcomes and assisted reproductive methods selection.

Sperm morphology is the best predictor of fertilization potential, and the critical predictive information for supporting assisted reproductive methods selection. Given its important predictive value and the declining reality of semen quality in recent years, the threshold of normal sperm morphology rate (NSMR) is being constantly corrected and controversial, from the 4th edition (14%) to the 5th version (4%). We retrospectively analyzed 4756 cases of infertility patients treated with conventional-IVF(c-IVF) or ICSI, which were divided into three groups according to NSMR: ≥14%, 4%–14% and <4%. Here, we demonstrate that, with decrease in NSMR(≥14%, 4%–14%, <4%), in the c-IVF group, the rate of fertilization, normal fertilization, high-quality embryo, multi-pregnancy and birth weight of twins gradually decreased significantly (P<0.05), while the miscarriage rate was significantly increased (p<0.01) and implantation rate, clinical pregnancy rate, ectopic pregnancy rate, preterm birth rate, live birth rate, sex ratio, and birth weight(Singleton) showed no significant change. In the ICSI group, with decrease in NSMR (≥14%, 4%–14%, <4%), high-quality embryo rate, multi-pregnancy rate and birth weight of twins were gradually decreased significantly (p<0.05), while other parameters had no significant difference. Considering the clinical assisted methods selection, in the NFMR ≥14% group, normal fertilization rate of c-IVF was significantly higher than the ICSI group (P<0.05), in the 4%–14% group, birth weight (twins) of c-IVF were significantly higher than the ICSI group, in the <4% group, miscarriage of IVF was significantly higher than the ICSI group. Therefore, we conclude that NSMR is positively related to embryo reproductive potential, and when NSMR<4% (5th edition), ICSI should be considered first, while the NSMR≥4%, c-IVF assisted reproduction might be preferred.

A new technique for analysis of human sperm morphology in unstained cells from raw semen.

Sperm morphology analysis is a fundamental component of semen analysis, but its real significance has been clouded by the plethora of techniques used for its evaluation. Most involve different fixation and staining procedures that induce artefacts. Herein we describe Trumorph (Proiser R+D, Paterna, Spain), a new method for sperm morphology analysis based on examination of wet preparations of spermatozoa immobilised, after a short 60°C shock, in narrow chambers and examined by negative phase contrast microscopy. A range of morphological forms was observed, similar to those found using conventional fixed and stained preparations, but other forms were also found, distinguishable only by the optics used. The ease of preparation makes the Trumorph a robust method applicable for the analysis of living unmodified spermatozoa in a range of situations. Subsequent studies on well-characterised samples are required to describe the morphology of spermatozoa with fertilising potential.

Effect of age and abstinence on semen quality: A retrospective study in a teaching hospital

Abstract Objective To elucidate the effect of age and sexual abstinence on semen quality (semen volume, total count, progressive motility, vitality and morphology). Methods A total of 730 semen samples were analyzed. Subjects were grouped according to the age (20-29, 30–34, 35–39 and 40–50) and abstinence (2–3, 4–5 and 6–7). Semen parameters were evaluated following WHO standard criteria. Results Analysis of 730 semen samples showed negative correlation of progressive motility ( r =-0.131, P r =-0.173, P r =-0.324, P H = 20.65, P H = 13.53, P H = 15.33, P U test confirmed the changes in semen volume, total count and vitality in paired grouping from 2–7 days ( P P Conclusions In the present study, age negatively affected progressive motility, vitality and morphology of human sperm. Semen samples showed intra varied results within WHO amended abstinence period.

Effects of combined antioxidant supplementation on human sperm motility and morphology during sperm manipulation in vitro

To evaluate the effect of antioxidant combinations on human sperm motility and morphologic normality during preparation and in vitro incubation to improve sperm quality for assisted reproductive technology (ART). Prospective study. Laboratory. Six fertile males, 21 to 25 years of age, after 3 days of sexual abstinence. Sperm retrieved from patients with normal fertility prepared and incubated in vitro in medium supplemented with taurine, cysteine, and/or glutathione. Motility indices, including motility (MOT), average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), and morphologic normality. Taurine supplementation during sperm preparation had no significant effects on motility or morphologic normality. However, when sperm was incubated for 24 hours under taurine-supplemented conditions, the rates of reduction in MOT, VAP, VCL, and VSL were statistically significantly decreased compared with taurine-free conditions. Morphologic normality was maintained, regardless of taurine treatment. The optimal taurine concentration for improving sperm motility and morphologic normality during preparation and in vitro incubation was 1 mM. Subsequently, combined treatment with 1 mM taurine, 1 mM cysteine, and 1 mM glutathione induced statistically significant synergistic effects on MOT, VAP, VCL, and VSL without any alteration of morphologic normality. Combined treatment with taurine, cysteine, and glutathione statistically significantly ameliorated the reduction in sperm motility during in vitro manipulation of human sperm.

Variability of Semen Parameters with Time in Placebo Treated Men

We describe the variability of semen parameters with time in normal men receiving placebo. We also report the impact of season and geographic region, among other variables, on these parameters. Data from the placebo arms of 5 randomized, controlled trials were pooled. All trials set minimum standards for semen parameters as an eligibility criterion for entry. Semen parameters examined include volume, density, motility, total count, total motile count and morphology. Mixed model repeated measure analysis was used for statistical analysis. Coefficients of variation for each semen parameter and the percent change from baseline were calculated. The mean within-subject coefficient of variation for each semen parameter ranged from a low of 10% to a high of almost 50%. The contribution of season and region to variability was negligible. The reduction in variability with an increasing number of samples per time point had decreasing returns beyond 2 samples. There was considerable variation in semen parameters with time in subjects who received placebo. Variation could not be attributed to season or region. We observed a general negative trend in semen parameters in this population selected for normal baseline semen parameters, which was likely due to the placebo response or to regression toward the mean.

Artificially selected human sperm morphology after swim-up processing

The swim-up technique is a clinical practice used to select highly motile sperm cells from patient ejaculates to use in assisted fertilization. The aim of this study was to investigate whether the length of different sperm-cell components is related to gamete function. Thus, we explored whether swim-up technique selects for longer sperm cells than mean sperm cells from unprocessed ejaculates. Sperm midpiece, tail endpiece, and total length were measured before and after the swim-up selection by means of contrast-phase and electron microscopy. Correlations between sperm dimensions, sperm motility, and sperm concentration were also investigated. Swim-up selected cells with longer midpiece compared with the unprocessed fractions (5.8 μm (CI 5.52–6.16 μm) vs. 5.3 μm (CI 4.97–5.61 μm), p &lt; 0.05) and shorter tail endpiece (7.8 μm (CI 7.11–8.44 μm) vs. 8.5 μm (CI 7.81–9.14 μm), p &lt; 0.05 after meta-analysis), whereas no effect of swim-up selection was detected on the total sperm cell length. Individuals producing high sperm concentrations had longer sperm midpiece than had men producing lower sperm concentrations. It is concluded that short sperm flagellar tips with long midpieces may be used as biomarkers in infertility therapy.

Evaluation of a modified rapid staining protocol for human sperm morphology assessment

Various staining methods exist for sperm morphology assessment. Some staining protocols require chemicals, use of fume hoods and multiple steps. This can create storage, space and time issues within the lab. Fortunately, methods are available for rapid spemr staining. However, prior studies suggest not all rapid approaches are suitable/accurate for morphology assessment utilizing Krueger Strict criteria. The objective of this study was to validate accuracy of a modified rapid staining method for sperm morphology.

Effects of reactive oxygen species from activated leucocytes on human sperm motility, viability and morphology

The accumulated data suggest that inflammation can increase the level of reactive oxygen species (ROS), which contribute to impaired sperm function and male infertility. Therefore, we propose that inflammation-mediated production of ROS in male and female reproductive tracts hinder sperm fertilisation. To test this hypothesis, phorbol myristate acetate (PMA) with polymorphonuclear leucocytes (PMNs) was applied to generate endogenous ROS. We evaluated the time-dependent effects of ROS on human sperm motility, viability and mitochondrial membrane potential (MMP). The results showed that after treatment with PMA and PMNs, the motility of human spermatozoa significantly decreased to 50% on Day 1 and 15% on Day 4 compared with that of the, respectively, negative controls (P = 0.012). The viability of human spermatozoa decreased on Day 4 of PMA + PMNs treatment (P = 0.028). The MMP of human spermatozoa significantly decreased from Day 2 to Day 4 in the PMA + PMN group compared with that of the controls (P = 0.019). Taken together, the 4-day cultivation approach provided an accurate evaluation of sperm quality, especially sperm motility and MMP. Our findings indicated that endogenous inflammation increased ROS levels, which might induce sperm oxidative damage. Additionally, sperm motility might be one of the earliest and most sensitive indicators of this damage.

In Vitro Effects of Tire Debris Organic Extracts on Human Spermatozoa

Background & objectives: Environmental pollutants have detrimental effects on the reproductive function of human being and of animals. This study was aimed at evaluating the effects of tire debris, a fraction of particulate matter derived from tire wear, on motility and morphology of human spermatozoa. Methods: spermatozoa from ten healthy men were incubated for 4 hours with 10, 50 and 75 μg/ml of tire debris organic extract (TDOE). Sperm motility and morphology were evaluated according to WHO guidelines; the presence of apoptosis and necrosis was assessed by the AnnexinV/Propidium iodide assay. Ultrastructure of sperm organelles was observed by transmission electron microscopy. Results: incubation with 10 � g/ml of TDOE did not affect sperm quality, except for the value of apoptosis (median: 8% vs. 6%; P<0.01). Sperm motility and sperm morphology percentages (P<0.01)decreased when sperm were treated with 50 and 75 μg/ml TDOE and the percentage of apoptotic and necrotic sperm (P<0.01) increased concomitantly. At the same concentrations TEM analysis showed an increased percentage of reacted and swollen acrosomes, disrupted chromatin and altered axonemes. Interpretation & conclusions: the organic part of tire debris is potentially harmful to spermatozoa, particularly at a concentration of 50-75 μg/ml, inhibiting motility and compromising morphological integrity. Nevertheless, these results cannot yet be extrapolated to the in vivo outcome due to the lack of studies focused on this issue.

Measurement and significance of sperm morphology

The measurement or evaluation and clinical significance of human sperm morphology has always been and still is a controversial aspect of the semen analysis for the determination of a male's fertility potential. In this review the background of the development of the evaluation criteria for sperm morphology will be discussed. Aspects of criticism on the strict criteria definition and use of the criteria for sperm morphology evaluation will be discussed as well as possible reasons for the decline in normal sperm morphology values and how we can compromise for this phenomenon resulting in the very low normal reference value as published in the 2010 WHO manual for the Examination and Processing of Human Semen. One of the possible solutions may be to give more attention to a limited number of abnormal sperm morphology categories and the inclusion of sperm morphology patterns. It is concluded in this review that if done correctly and with care and with strict application of existing guidelines as outlined in the 2010 WHO manual, sperm morphology measurement still has a very important role to play in the clinical evaluation of male fertility potential.

Human normal sperm morphology rate and in vitro fertilization outcome

To determine the effect of sperm morphology on fertilization rate, cleavage rate,and clinical pregnancy rate, which may help to choose a best protocol before assisted reproductive technology (ART). We a retrospectively surveyed semen data of 1369 ART patients from January to December 2008 in our hospital. We divided sperms from 751 patients into 4 groups based on the sperm morphology rate: ≤5.0% (normal), 5.1%-10.0%, 10.1%-15.0%, and >15.0%. In vitro fertilization (IVF) was used for 490 patients or intracytoplasmic sperm injection (ICSI) for 261. We also divided sperms from 618 donors into 4 groups based on the sperm morphology rate: 14.0%-17.0%(normal), 17.1%-19.0%, 19.1%-23.0%, and >23.0%. Only IVF was used for these donors. The fertilization rate, cleavage rate, and clinical pregnancy rate among each group were analyzed and discussed. During the IVF(patients or donors) cycle, there was a significant difference among the 4 groups in the fertilization rate and clinical pregnancy rate (P<0.05), but there was no difference in the cleavage rate (P>0.05). In the ICSI cycle, there was no significant difference among the 4 groups in the fertilization rate, clinical pregnancy rate, and cleavage rate (P>0.05). Sperm morphology affects the outcome of IVF. There is no significant influence of sperm morphology on the outcome of ICSI. ICSI is an optimal alternative for patients with abnormal morphological sperms in ART.

IMSI : indication, résultats et réflexions

The presence of nuclear vacuoles in human sperm decreases pregnancy rates. Intracytoplasmic morphologically selected sperm injection (ISMI) increases pregnancy rate rather than ICSI after real time fine morphology of motile human sperm (MSOME). However, the exact indications of IMSI are on debate.