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The concentration of nitrite in seminal plasma does not correlate with sperm concentration, sperm motility, leukocytospermia, or sperm culture.

OBJECTIVE: To correlate the concentration of nitrite (the stable metabolite of nitric oxide) in seminal plasma with sperm number and motility, leukocytospermia, and sperm culture. DESIGN: Prospective study. SETTING: Academic research institution. PATIENT(S): Seventy normozoospermic or dyspermic men enrolled in an artificial insemination/in vitro fertilization program. INTERVENTION(S): Semen samples (n = 70) were checked for sperm concentration, total sperm count, sperm motility, seminal leukocyte concentration, and sperm culture; similarly, the concentration of nitrite in seminal plasma was measured by Griess reaction. MAIN OUTCOME MEASURE(S): Measurement of nitrite concentration in seminal plasma and its correlation with sperm concentration, total sperm count, sperm motility, leukocytospermia, and sperm culture. RESULT(S): The concentration of nitrite in seminal plasma does not correlate with sperm concentration, total sperm count, or with the proportion of immotile or rapid-forward motile spermatozoa. Moreover, the concentration of nitrite in seminal plasma is not significantly increased when sperm culture is positive, nor does it correlate with leukocyte concentration in semen. CONCLUSION(S): Our results do not support the hypothesis that in vivo nitric oxide synthesis affects sperm function; alternatively, our results could suggest that nitrite in the seminal plasma is not a sensitive marker of in vivo nitric oxide synthesis.

Fertilization in Vitro↗

Extragonadal sperm reserves, sperm-depletion rates, numbers of sperm per mating, and fertility with successive matings by intact or unilaterally vasectomized rats.

Because of the high rates of sperm production and large extragonadal sperm reserves of sexually rested male rats, mating trials are insensitive for detecting test-induced alterations in sperm production rates. Mating trials might be more sensitive if, independently of any experimental treatments under study, the number of sperm per mating was closer to the minimum requirements for normal fertility. The present study was undertaken to assess the impact of unilateral vasectomy and/or matings with up to three females in succession, for 1 hour each, on the number of sperm per mating and fertility, in comparison to corresponding values for males allowed unlimited matings with a receptive female overnight. Unilateral vasectomy did not affect sperm production, extragonadal sperm reserves, or removal of contralateral sperm during ejaculation (P > 0.05) but caused a 50% decrease in sperm numbers per mating. Sperm output, judged from numbers of residual extragonadal sperm in unmated and mated males, was excessive (290 x 10[6]) during conventional overnight mating with intact males and during the first and second hours of restricted mating (105 and 184 x 10(6) respectively, for intact males; one-half of these amounts for unilaterally vasectomized males). In contrast, sperm output during the third successive mating was minimal (nonmeasurable) but adequate, since pregnancy rates were similar for females mated first, second, or third in succession (P > 0.05). Since successive matings reduce the number of sperm per mating by natural methods, this approach may enhance the sensitivity of mating tests when applied for assessing the potential effects of experimental treatments on sperm production.

Animals↗

Evolution of sperm size in nematodes: sperm competition favours larger sperm.

In the free-living rhabditid nematode Caenorhabditis elegans, sperm size is a determinant of sperm competitiveness. Larger sperm crawl faster and physically displace smaller sperm to take fertilization priority, but not without a cost: larger sperm are produced at a slower rate. Here, we investigate the evolution of sperm size in the family Rhabditidae by comparing sperm among 19 species, seven of which are hermaphroditic (self-fertile hermaphrodites and males), the rest being gonochoristic (females and males). We found that sperm size differed significantly with reproductive mode: males of gonochoristic species had significantly larger sperm than did males of the hermaphroditic species. Because males compose 50% of the populations of gonochoristic species but are rare in hermaphroditic species, the risk of male-male sperm competition is greater in gonochoristic species. Larger sperm have thus evolved in species with a greater risk of sperm competition. Our results support recent studies contending that sperm size may increase in response to sperm competition.

Animals↗

Relative testis size and sperm morphometry across mammals: no evidence for an association between sperm competition and sperm length.

Understanding why there is extensive variation in sperm form and function across taxa has been a challenge because sperm are specialized cells operating at a microscopic level in a complex environment. This comparative study collates published data to determine whether the evolution of sperm morphometry (sperm total length and separate component dimensions) is associated with sperm competition (when different males' sperm mix and compete for a female's ova) across 83 mammalian species. We use relative testes mass as an indicator of the intensity of sperm competition across taxa: relative investment into testes is widely accepted to predict the level of sperm competition that a species or population endures. Although we found evidence for positive associations between relative testes mass (controlling for allometry) and sperm morphometry across 83 mammalian species, these relationships were phylogenetically dependent. When we appropriately controlled for phylogenetic association using multiple regression within a phylogenetic framework, there was no relationship between relative testes mass and sperm length across mammals. Furthermore, we found no evidence for associations between relative testes mass and sperm head, mid-piece or flagellar lengths, nor was there a relationship with mid-piece or mitochondrial volumes. Results, therefore, indicate that sperm competition does not select for longer or shorter sperm across mammals, and alternative forces selecting on sperm form and function are discussed.

Animals↗

Novel association between sperm reactive oxygen species production, sperm morphological defects, and the sperm deformity index.

OBJECTIVE: To examine the relationship between sperm reactive oxygen species (ROS) production and sperm morphology in a group of infertile men and healthy fertile donors. DESIGN: A prospective clinical study. SETTING: Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio, and the Reproductive Medicine Unit, Liverpool Women's Hospital, United Kingdom PATIENT(S): Thirty-nine infertile men and 13 healthy fertile donors (control). INTERVENTION(S): Standard semen analysis, seminal leukocyte concentration, assessment of sperm morphology, and measurement of sperm ROS production. MAIN OUTCOME MEASURE(S): Levels of sperm ROS production, percentages of different sperm morphological abnormalities, and the sperm deformity index (SDI) scores. RESULT(S): A significant negative correlation was observed between sperm ROS production and the proportion of sperm with normal morphology and borderline morphology. Reactive oxygen species production was positively correlated with the proportion of sperm with amorphous heads, damaged acrosomes, midpiece defects, cytoplasmic droplets, tail defects, and SDI scores. Logistic regression analysis identified a two-variable model including SDI and percentage sperm motility, which correctly identified 84% of individuals with high seminal ROS and 85% of individuals with low seminal ROS. The model had an overall accuracy of 85%. CONCLUSION(S): The standard semen analysis to assess sperm motility, sperm morphology, and the SDI scores is a useful tool in identifying infertile men with high seminal ROS in infertility clinics where facilities for measuring levels of seminal ROS are not available.

Female↗

Sperm nuclear chromatin normality: relationship with sperm morphology, sperm-zona pellucida binding, and fertilization rates in vitro.

OBJECTIVE: To study whether the results of tests of sperm chromatin and deoxyribonucleic acid (DNA) normality are related to fertilization rates in vitro. DESIGN: Normal morphology, nuclear maturity determined by acidic aniline blue stain, and DNA normality determined by acridine orange fluorescence of sperm in insemination medium and the number of sperm bound to the zona pellucida (ZP) of the oocytes that had failed to fertilize in vitro were determined. The relationship between sperm test results and fertilization rates were analyzed by logistic regression. SETTING: Samples were obtained from patients undergoing in vitro fertilization (IVF) treatment. RESULTS: The number of sperm bound to the ZP, the percentage of sperm with normal morphology, and the percentage of sperm with normal DNA were the most significant factors related to fertilization rates in vitro. In patients with normal morphology > or = 15% or with > 10 sperm bound per ZP, the percentage of sperm with normal DNA, the number of sperm bound to the ZP, and motility grade were significantly related to IVF rates. CONCLUSION: In patients with normal morphology > or = 15%, failure of fertilization may be because of defects of sperm-ZP binding or abnormal DNA. Assessment of DNA normality of motile sperm in the insemination medium may aid prediction of fertilization rates in addition to normal morphology and sperm-ZP binding.

Acridine Orange↗

Evidence favoring sperm selection over sperm competition in the interaction between human seminal plasma and sperm motility in vitro.

The effect on sperm motility of sperm-sperm and sperm-seminal plasma interactions was studied among homologous and heterologous sperm. There were no significant interactions between sperm in vitro, but it was found that seminal plasmas of different donors have different effects on sperm motility, and different sperm react differently to the same seminal plasma. Sperm showed higher motility in a pure physiological solution than when mixed with seminal plasma, even if the plasma and sperm came from the same donor. Various plasma components are responsible for this modulation of sperm motility. It would appear that large numbers of sperm are adaptive, among other things, because they are involved in sperm selection.

Adolescent↗

Evaluation of computerized analysis of sperm movement characteristics and differential sperm tail swelling patterns in predicting human sperm in vitro fertilizing capacity.

Multivariate discriminant analysis was used to evaluate the usefulness of computer image analysis of sperm movement characteristics and differential patterns of sperm tail swelling after hypoosmotic treatment for predicting the human sperm in vitro fertilizing capacity assessed by the zona-free hamster egg penetration assay. Fifty-five semen samples, mostly normospermic, from untreated infertility clinic patients were analyzed. The % normal sperm morphology, linearity of seminal sperm movement, seminal sperm head beat frequency, mean and maximum amplitudes of lateral head displacement, and hypoosmotic sperm tail swelling patterns c, d and f were selected by multivariate discriminant analysis to be capable of discriminating the samples exhibiting the presence or the absence of sperm in vitro fertilizing capacity. The % total sperm tail swelling did not give additional information about in vitro fertilizing capacity. These preliminary data suggest that computer image analysis of sperm movement characteristics and differential evaluation of hypoosmotic sperm tail swelling might be useful for the prediction of human sperm fertility. Further prospective studies are necessary to validate their predictive functions.

Evaluation Studies as Topic↗

Sperm maturity and treatment choice of in vitro fertilization (IVF) or intracytoplasmic sperm injection: diminished sperm HspA2 chaperone levels predict IVF failure.

OBJECTIVE: To reexamine whether low sperm HspA2 ratios (formerly sperm CK-M ratio) are predictive for failure to cause pregnancies by in vitro fertilization (IVF) and to explore whether there are other male or female factors that may be predictive for IVF pregnancy outcome. DESIGN: Retrospective, cohort study. SETTING: University-based IVF program. PATIENT(S): In 119 IVF cycles, three patient groups were studied: 25 men had a <10% sperm HspA2 ratio and a lack of pregnancies (HS <10% group), 50 men had a >10% sperm HspA2 ratio and no pregnancies (HS >10%NP group), and another 44 couples had a >10% sperm HspA2 ratio but did achieve pregnancies (HS >10%P group). INTERVENTION(S): Sperm HspA2 ratio determinations within 1 year of the IVF cycles and analysis of male and female IVF cycle parameters. MAIN OUTCOME MEASURE(S): Sperm HspA2 ratio determinations within 1 year of the IVF cycles and analysis of male and female IVF cycle parameters. RESULT(S): In the three groups, male and female ages, number and maturation level of oocytes, and morphology of embryos were similar. In the HS < 10% group, mean sperm concentration and motility were close to normal, the fertilization and cleavage rates were lower, and cycles without any oocyte fertilization were higher. These parameters were similar in the two HS > 10% groups. The receiver operating characteristic curve in men with sperm HspA2 ratios of <17% (diminished and borderline sperm maturity) provided a cutoff value of 10.84% HspA2 ratio with a 100% positive predictive value for failure to achieve pregnancy, whether the men were oligospermic or normospermic. CONCLUSION(S): HspA2 ratios of <10% in the diminished sperm maturity range predict the failure to cause pregnancies by IVF. Thus, IVF should be bypassed in favor of ICSI. The occurrence of pregnancy with ICSI depends on the maturity of sperm selected, and it may not be as likely as in other indications for ICSI.

Adult↗

Comparison of efficacy of two techniques for testicular sperm retrieval in nonobstructive azoospermia: multifocal testicular sperm extraction versus multifocal testicular sperm aspiration.

To compare the efficacy of 2 sperm-retrieval procedures, testicular sperm extraction (TESE) and testicular sperm aspiration (TESA), during the same procedure using the same subjects as their own controls. The presence of mature testicular sperm cells and motility were evaluated in 87 men with nonobstructive azoospermia (NOA) by means of multifocal TESE and multifocal TESA, which were performed during the same procedure using the same subjects as their own controls. Sperm cells were recovered by TESE in 54 cases, but by TESA in only 36 cases. There were significantly more cases (n = 20) in which sperm cells were recovered by TESE only, compared with 2 cases in whom cells were recovered by TESA only (McNemar's test, P < .001). The mean number of locations in each testis in which sperm cells were detected was significantly higher in the TESE group. In significantly more cases (n = 27), motility was observed in TESE material only, compared with 3 cases in which motility was present in material extracted by TESA only (McNemar's test, P < .001). Mean number of locations in each testis with motile sperm cells was significantly higher in the TESE group. The TESE procedure yielded significantly more sperm cells, as was also reflected by the difference in number of straws with cryopreserved sperm. This comparative prospective clinical study revealed that multifocal TESE is more efficient than multifocal TESA for sperm detection and recovery in men with NOA and should be the procedure of choice for sperm retrieval for them.

Adult↗

Analysis of sperm quality in recombinant inbred mouse strains: correlation of sperm head shape with sperm abnormalities and with the incidence of supplementary spermatozoa in the perivitelline space.

Recombinant inbred strains were developed from reciprocal crosses between two inbred strains of mice (CBA and KE) differing in sperm head shape, proportion of normal sperm heads (CBA, 95%; KE, 78%) and fertilization efficiency (CBA, 100% of fertilized ova; KE, 72%), to determine whether the indices of sperm morphology and function were correlated. The following parameters were analysed in recombinant inbred and progenitor strains: index of sperm head shape (head width in the middle of its length/head length), percentage of abnormal sperm heads, percentage of spermatozoa with progressive movements, efficiency of penetration of hyaluronic acid polymer (Sperm Select) and percentage of fertilized ova after mating males from the tested strains with females from an outbred stock. For each investigated character, recombinant inbred strains, recombinant inbred EXCB and CBXE, could be divided into at least three categories: KE-like, CBA-like and intermediate, suggesting that in each case a minimum of two genes was involved. Recombinant strains derived from the reciprocal crosses of progenitor strains differed only with respect to the proportion of abnormal sperm heads, showing the involvement of the Y chromosome in determining this character. Penetration into Sperm Select was significantly correlated both with fertilization efficiency and sperm motility, while correlation with the proportion of normal spermatozoa did not reach the level of significance. However, there was a significant negative correlation of both sperm abnormalities and the incidence of supplementary spermatozoa in the perivitelline space with the index of sperm head shape.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Synchronous sperm retrieval and sperm washing in an intracytoplasmic sperm injection cycle in an azoospermic man who was positive for human immunodeficiency virus.

OBJECTIVE: To present the first reported case of synchronous sperm retrieval followed by sperm washing before an intracytoplasmic sperm injection (ICSI) cycle in an HIV-positive azoospermic man. DESIGN: Case report. SETTING: Assisted reproduction center. PATIENT(S): A 40-year-old HIV-positive man with obstructive azoospermia due to vasal aplasia. INTERVENTION(S): Synchronous sperm retrieval, sperm washing, nucleic acid-based sequence amplification testing, and intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): Successful sperm retrieval sufficient for sperm washing and fertilization. RESULT(S): Sufficient quantity of spermatozoa for washing was obtained at epididymal aspiration. After the wash, HIV ribonucleic acid (RNA) was undetectable with nucleic acid-based sequence amplification testing, enabling injection of oocytes collected after routine gonadotropin superovulation. Of seven oocytes collected from the 39-year-old woman partner, six were injected and five fertilized (83%). Three embryos were transferred on day 2. The pregnancy test was negative on this occasion. CONCLUSION(S): This case demonstrates that sperm washing can be applied in cases of sperm retrieval where sperm volume and density is low, allowing the treatment of azoospermic HIV-positive men.

Adult↗

Goat sperm membrane: lectin-binding sites of sperm surface and lectin affinity chromatography of the mature sperm membrane antigens.

The cell surface glycoproteins of goat epididymal maturing spermatozoa have been investigated using lectins as surface probes that interact with specific sugars with high affinity. Concanavalin A (ConA) and wheat-germ agglutinin (WGA) showed high affinity for mature cauda epididymal sperm agglutination, whereas RCA2, kidney beans lectin and peanut agglutinin caused much lower or little agglutination of the cells. The mature sperm exhibited markedly higher efficacy than the immature caput epididymal sperm for binding both ConA and WGA, as evidenced by sperm agglutination and the binding of the fluorescence isothiocyanate (FITC)-labelled lectins. FITC-ConA binds uniformly to the entire mature sperm surface whereas FITC-WGA binds to the acrosomal cap region of the head. The FITC-RCA2 mainly labelled the posterior head of mature cauda sperm. However, no WGA-specific glycoprotein receptors could be detected in sperm plasma membrane (PM) by WGA-Sepharose affinity chromatography. The data implied that the epididymal sperm maturation is associated with a marked increase in the ConA/WGA receptors and that WGA receptors may be glycolipids rather than glycoproteins. Analysis of the ConA receptors of cauda sperm PM identified by ConA-Sepharose affinity chromatography and subsequent resolution in SDS-PAGE demonstrated the presence of five glycopolypeptides of different concentrations (98, 96, 43, 27 and 17 kDa) of goat sperm membrane. The immunoblot of these ConA-specific glycopeptides with anti-sperm membrane antiserum showed that 98- and 96-kDa receptors are immunoresponsive.

Agglutination↗

Factors related to successful sperm microinjection of hamster eggs: The effect of sperm species, technical experience, needle dimensions, and incubation medium on egg viability and sperm decondensation following microinjection.

A series of experiments was designed to identify factors in a sperm microinjection system that could influence egg viability and decondensation of sperm nuclei after microinjection. Egg viability and sperm decondensation rates were not different among eggs microinjected with rodent sperm. The microinjection of ram sperm required a larger diameter needle for injection, which resulted in low egg viability and sperm decondensation in the first 3 mo of the study but improved greatly after 9 mo of technical experience. The degree of technical experience (3 vs 9 mo) also improved (P<0.05) egg viability after microinjection with rodent sperm; however, the rate of sperm decondensation remained unaffected. Altering the dimensions of the injection needle from a tapered needle barrel to a more uniform needle barrel increased egg viability from 61 to 96% and sperm decondensation from 3 to 27%. The use of medium 199 for incubating microinjected eggs further increased (P<0.05) the percentage of eggs containing decondensed sperm nuclei (52%) compared to eggs incubated in Holmes defined medium (28%). By altering the dimensions of the injection needle, by selecting an appropriate incubation medium, and by gaining technical experience in microinjection, the efficiency of a sperm microinjection system was improved for both rodents and domestic animals.

Journal Article↗

The Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI.

The Beltsville sperm sexing technology is currently the only effective means of altering the sex ratio of offspring in livestock. The method is based on the flow-cytometric separation of X- and Y-chromosome-bearing sperm based on X/Y DNA content difference. It is an effective means of producing progeny of predetermined sex in cattle, swine, sheep, and laboratory animals. The method involves treating sperm with a DNA-binding fluorochrome, Hoechst 33342, and flow-cytometrically sorting them into separate X and Y populations that can subsequently be used for surgical intratubal or intrauterine insemination, deep-uterine insemination, regular artificial insemination in some cases, in vitro fertilization to produce sexed embryos for transfer, and intracytoplasmic sperm injection of ova. Skewed sex ratios of 85 to 95% of one sex or the other have been repeatably achieved in most species. The method has been used worldwide to produce several hundred morphologically normal animal offspring of the predicted sex. It has also been validated in the laboratory using DNA reanalysis of the sorted sperm populations and by fluorescence in situ hybridization and PCR of individual sperm. We developed a new orienting nozzle that we have fitted to both conventional and high-speed cell sorters that have been modified for sperm sorting. Recently we completed the adaptation of the new orienting nozzle to a Cytomation MoFlo high-speed cell sorter modified for sperm. This adaptation of the nozzle has increased the overall production rate of sorted X and Y sperm from about .35 million/h to 5 or 6 million sperm/h (each population). Calves have been born from cows artificially inseminated using conventional technique and sexed sperm. In addition, numerous litters of pigs have been born after transfer of embryos produced from X or Y sorted sperm.

Animals↗

Evaluation of rat sperm by flow cytometry: simultaneous analysis of sperm count and sperm viability.

In this study, we conducted a simultaneous analysis of sperm count and viability in rats by flow cytometry (FCM). Epididymal fluids were taken from the caudal epididymis of 12 to 13 week-old Sprague-Dawley rats. The fluids were weighed and mixed with Dulbecco's phosphate buffered saline (D-PBS). Propidium iodide, which can stain only dead sperm, was used to distinguish viable and dead sperm. The sperm count and viability analyzed by FCM were 1.28 x 10(6)/mg and 78.0%, respectively. These values were consistent with the corresponding values (1.39 x 10(6)/mg and 81.0%) that were directly determined microscopically in the fluids of the same sample. In addition, when the original mixture containing sperm was diluted two times and four times with D-PBS, or was diluted two times with D-PBS containing only killed sperm, the sperm count and viability determined by FCM also correlated well with the sperm count (r = 0.96, P < 0.01) and sperm motility (r = 0.99, P < 0.01) by direct microscopic observation, respectively. In conclusion, the present flow cytometric analysis would be practical for the simultaneous determination of sperm count and viability in rat epididymal fluids.

Animals↗

Murine sperm-zona binding, a fucosyl residue is required for a high affinity sperm-binding ligand. A second site on sperm binds a nonfucosylated, beta-galactosyl-capped oligosaccharide.

An essential initial step in murine fertilization is the binding of acrosome-intact sperm to specific O-linked oligosaccharides on zona pellucida glycoprotein 3. While there is agreement on the primary role of O-linked glycans in this process, there is a lack of consensus on both the terminal monosaccharide(s) required for a functional sperm binding site and the corresponding protein on the sperm cell surface that recognizes this ligand. Much current debate centers on an essential role for either a terminal N-acetylglucosaminyl or, alternatively, a terminal alpha-galactosyl residue. To gain insight into the terminal saccharides required to form a functional sperm-binding ligand, dose-response curves were generated for a series of related tri- and tetrasaccharides to evaluate their relative effectiveness to competitively inhibit the in vitro binding of murine sperm to zona pellucida-enclosed eggs. A GlcNAc-capped trisaccharide, GlcNAc beta 1,4GlcNAc beta 1,4GlcNAc,was inactive (1-72 microM range). In contrast, a beta 4-galactosyl-capped trisaccharide (Gal beta 1,4GlcNAc beta 1, 4GlcNAc) and an alpha 3-galactosyl-capped trisaccharide (Gal alpha 1,3Gal beta 1,4 GlcNAc) inhibited sperm-zona binding with low or moderate affinity (ED50 = 42 microM and 5.3 microM, respectively). The addition of an alpha 3-fucosyl residue to each of these two competitive inhibitors, forming Gal beta 1,4[Fuc alpha 1,3] GlcNAc beta 1,4GlcNAc or Gal alpha 1,3Gal beta 1, 4[Fuc alpha 1,3]Glc NAc, resulted in ligands with 85- and 12-fold higher affinities for sperm, respectively (ED50 = 500 and 430 nM). Thus, the presence of a fucosyl residue appears to be obligatory for an oligosaccharide to bind sperm with high affinity. Last, mixing experiments with pairs of competitive inhibitors suggest that murine sperm-zona binding is mediated by two independent oligosaccharide-binding sites on sperm. The first (apparently high affinity) site binds both the alpha 3-galactosyl-capped trisaccharide and the two fucosylated tetrasaccharides. The second (apparently low affinity) site binds a nonfucosylated beta-galactosyl-capped trisaccharide.

Acrosome↗

Prediction of post-thaw sperm motility and sperm cryosurvival rate using the pre-freeze sperm parameters.

To determine the relationships between pre-freeze semen variables and cryosurvival rate and post-thaw motility and examine whether they have any predictive value for the cryosurvival rate and post-thaw sperm motility, conventional semen analysis, supravital staining for sperm viability and hypo-osmotic swelling test were performed on 50 semen samples before cryopreservation. Thawed semen samples were examined for post-thaw sperm motility and cryosurvival rate. Significant correlations were observed between post-thaw sperm motility and several pre-freeze semen variables, such as, hypo-osmotic swelling test, pre-freeze sperm motility and sperm viability. In a stepwise regression analysis, an accurate prediction of post-thaw sperm motility (R = 0.826) was obtained using a multiple regression equation incorporating 3 variables including hypo-osmotic swelling test, pre-freeze sperm motility and sperm concentration. In conclusion, a set of criteria have been identified that accurately predicts post-thaw sperm motility and which place particular emphasis on hypo-osmotic swelling test. Conventional semen analysis and hypo-osmotic swelling test are simple and effective assays for the prediction of post-thaw motility.

Cryopreservation↗