Prognostic value of an automated sperm analysis in IVF or insemination therapy

Summary. During the course of sterility treatment semenograms of 271 IVF and 316 insemination patients were carried out by two automated semen analysers. The parameters of these analyses were correlated to pregnancies resulting from the treatment. Semen samples were analysed in the ejaculate and after swim-up preparation. In addition, the swim-up suspension of IVF patients was measured again 18 h after sperm preparation. Patients were divided into three groups: (1) couples who achieved a pregnancy, (2) couples who did not achieve a pregnancy, and (3) IVF patients with no fertilization of the oocytes. Because of large standard deviations in the quality of ejaculates, the results in the IVF group show no significant differences in the semen parameters of husbands of pregnant and non-pregnant women. In contrast husbands of women with no fertilization have a significantly lower sperm motility. After swim-up preparation these differences no longer occur. A further measurement, taken 18 h later, reveals that there are no differences in the sperm parameters between the pregnant and non-pregnant group. However, the semen quality in the group with no fertilization is significantly reduced. The results of the insemination patients are similar to those of the IVF group. Thus, the results from automated sperm analysers cannot replace either the microscopic or biochemical analysis of an ejaculate and, moreover, cannot be used as prognosis for the fertilization capacity of sperms or a following pregnancy.     

Morphology of seminal and swim-up spermatozoa and the outcome of in vitro fertilization and embryo transfer

Tubal infertility was treated by in vitro fertilization-embryo transfer (IVF-ET) in 112 couples. Twenty-eight pregnancies were obtained in 140 treatment cycles. Couples are accepted for treatment in our IVF-ET programme if previous semen samples fulfil the inclusion criteria: ejaculate volume greater than 1.5 ml, concentration of spermatozoa greater than 15 x 10(6) ml-1, greater than 40% motile spermatozoa, and greater than 25% spermatozoa with normal morphology. In order to determine to which extent IVF-ET treatment results are influenced by sperm morphology, within this selected group of patients, we have retrospectively analysed the data from both original semen samples and swim-up preparations. The sperm morphology was not related to the outcome of treatment in terms of fertilization (ovum cleavage rate), early embryo development, or pregnancy. Nor was any relationship detected between early embryo development or pregnancy and the degree of improvement in morphology resulting from the swim-up procedure. However, if improvement in morphology by swim-up was high, ovum cleavage rate was low. Sperm morphology within the limits set by our inclusion criteria could not predict the outcome of IVF-ET treatment. It is further concluded that the presence of abnormal spermatozoa at the site of fertilization may be without harm if only the number of normal sperms is high enough.     

Changes in human sperm chromatin stability during preparation for in-vitro fertilization

This study was designed to define the effects of sperm preparation on sperm chromatin stability in relation to in-vitro fertilization (IVF). Semen samples used for IVF-embryo transfer (ET) in the treatment of infertility due to tubal factors were studied. Cases with semen variables below reference limits in previous samples were excluded. Sperm were prepared by a swim-up technique employing either of two different tissue culture media, Ham's F-10 or Earle's balanced salt solution. Sperm chromatin stability was tested by exposure both to sodium dodecyl sulphate (SDS) only and SDS together with a zinc-chelating agent, disodium ethylene diamine tetraacetate (SDS-EDTA). Sperm head swell scores were defined under different experimental conditions and the relationship to sperm motility, morphology, fertilization rate and pregnancy occurrence was tested. No differences were seen between the chromatin stability of sperm from the original sample and that after swim-up preparation, neither immediately after completion of the swim-up procedure, nor at the time of insemination of ova. With time, the chromatin became more stable, which occurred to a similar extent both in the original sample and in swim-up preparations using Ham's F-10. Otherwise, sperm chromatin stability was unaffected by either of the two media used for swim-up. At higher incubation temperatures, decondensation in SDS was enhanced. Altogether, no correlation was found between sperm chromatin stability or enhancement of decondensation by temperature and the success of IVF treatment expressed in fertilization rates or pregnancies. The results are reassuring in that only small changes in sperm chromatin stability occurred during the preparation for IVF. As long as semen of presumably good quality is used, these changes in chromatin stability do not seem to be of clinical importance.     

不孕症患者宫腔内人工授精精子优选方法分析

目的:比较分析精子不同优选方法与宫腔内人工授精(IUI)成功率的相关性,以提高IUI妊娠率。方法:回顾性分析本院生殖中心门诊不孕症患者452例671个IUI周期,按精子优选方法分为3组:含5%人血清白蛋白(HSA)的Earle's液上游法、SpermR inse上游法、SupraSperm密度梯度离心法。比较各组患者的处理前后精子密度与活动率及临床妊娠结局。结果:3种精子优选方法处理后精子活动率及a+b级精子百分率显著增加(P<0.01)。含5%HSA的Earle's液上游法221个IUI周期,妊娠21个周期,周期临床妊娠率9.5%;SpermR inse上游法215个IUI周期,妊娠34个周期,周期临床妊娠率15.8%,与含5%HSA的Earle's液上游法比较有显著性差异(P<0.05);SupraSperm密度梯度离心法235个IUI周期,妊娠34个周期,周期临床妊娠率14.5%,与含5%HSA的Earle's液上游法比较无统计学差异(P>0.05)。结论:SpermR inse上游法的IUI临床妊娠率明显高于含5%HSA的Earle's液上游法,适用于各类不孕症患者。SupraSperm密度梯度离心法也具有较高的周期临床妊娠率,洗涤效果好,操作便捷高效,尤其适用于含炎症细胞、死精子和畸形精子等不良成分较多的精液。     

Migration sedimentation technique as a predictive test for the fertilizing capacity of spermatozoa in an in-vitro fertilization programme

The migration-sedimentation technique (MST) has been proposed as a means of separating high quality motile spermatozoa. The present study was conducted in order to evaluate whether sperm performance following separation by MST predicts their fertilizing capacity in an in-vitro fertilization (IVF) programme. Ninety semen specimens were analysed for use in an IVF-embryo transfer (ET) programme. Each specimens was divided into two parts: one was processed in the IVF programme and was used after sperm swim-up separation for insemination of human ova. The other aliquot (0.2 ml) was separated by MST, and the sperm then characterized by their concentration, motility, degree of motility and morphology. Sperm characteristics after separation by MST were then correlated with the results of the IVF-fertilization rates. In 79 of 90 IVF-ET cycles, at least one oocyte was fertilized. All post-MST sperm characteristics were significantly higher in cycles with fertilizations compared to IVF cycles without fertilization. A larger percentage of the total motile spermatozoa were recovered after MST in semen specimens with fertilization, compared to semen specimens without fertilization (39.9 +/- 3.6 and 20.6 +/- 6.6%, respectively; P < 0.05). This value was correlated with the percentage of fertilized oocytes (r = 0.24; P < 0.02). More IVF cycles with fertilizations were recorded in cases in which the recovery of motile sperm was > 25% (P < 0.005), or when more than 1.5 x 10(6) motile spermatozoa were recovered after MST (P < 0.0001). As sperm characteristics after MST correlated significantly with their fertilizing capacity, the MST test could be used in evaluation of the fertilizing capacity of spermatozoa.     

上游处理前后正常形态精子百分率与体外受精-胚胎移植结局的关系

目的:探讨取卵当日上游处理前后精子形态与体外受精-胚胎移植(IVF-ET)治疗结局的关系。方法:选取因单纯输卵管因素不孕而拟行IVF-ET治疗的夫妇94对。使用Kruger标准评价取卵当日上游处理前后的精子形态。以正常精子形态百分率10%为界,分别将上游处理前后的精子分成两组:B1组上游处理前正常形态≥10%,B2组<10%;A1组上游处理后正常形态≥10%,A2组<10%。分别比较B1和B2组以及A1和A2组的IVF-ET治疗结局。结果:上游处理前,B1组的受精率、卵裂率、优质胚胎率、妊娠率和胚胎种植率分别为(72.90±4.23)%、(95.20±2.61)%、(23.35±5.19)%、39.58%、18.35%;B2组分别为(71.33±5.10)%、(95.71±2.88)%、(20.18±6.15)%、41.86%、21.28%;各项指标两组之间差异均无统计学意义(P>0.05)。上游处理后,A1组受精率、卵裂率、优质胚胎率、妊娠率、胚胎种植率分别为(72.72±3.35)%、(95.64±2.04)%、(24.39±4.57)%、50.00%、23.87%;A2组分别为(70.27±8.82)%、(94.82±4.94)%、(13.45±7.39)%、9.52%、6.25%;两组之间受精率及卵裂率差别无统计学意义(P>0.05),但A1组的优质胚胎率、妊娠率及种植率明显高于A2组(P<0.05)。结论:取卵当日经上游处理后的正常精子形态百分率对IVF-ET结局有较好的预测价值。     

精子综合优选法配合宫腔内联合宫颈管内人工授精疗效分析

目的 探讨运用综合方法 优选精子,并采用宫腔内联合宫颈管内人工授精,对不育患者的临床治疗效果.方法 选取2005年1月至2009年12月期间在本院进行夫精人工授精（AIH）治疗的患者276例,共治疗565周期,随机分为对照组（n=138）和治疗组（n=138）.对照组采用直接或洗涤1次后上游法优选的精子用于宫腔内人工授精280周期;治疗组综合采用直接或洗涤1次后上游法优选的精子,用于宫腔内人工授精,将上游后剩留下的精液,用直接离心法优选得到的精子用于宫颈管内人工授精285周期,观察两组的治疗成功率.结果 对照组妊娠42例,妊娠率30.43 %,治疗周期妊娠率15.00%;治疗组妊娠73例,妊娠率52.89 %,治疗周期妊娠率25.61%,两组的妊娠率及周期妊娠率比较差异有统计学意义（P〈0.01）.结论 本院采用的改良AIH是一种高效、安全的人工授精方法.     

Intrauterine insemination with double ejaculate compared with single ejaculate in male factor infertility: a pilot study

The aim of this study was to compare the efficiency of using a double ejaculate with the efficiency of using a single ejaculate for intrauterine insemination in male subfertility. Eligibility for the study was a total motile sperm count between 1 x 10(6) and 5 x 10(6) on postwash sperm analysis. Thirty-nine couples were randomized to the study group, while another 50 couples were randomized to serve as the control group. Males in the study group were asked to produce a second semen sample within 2 hours of the first sample on the day of insemination. In the study group, the mean total motile sperm count in the first sample was 3.83 x 10(6) (SD +/- 0.85 x 10(6); range 1.1-4.9) and 3.99 x 10(6) (SD +/- 0.72 x 10(6); range 0.9-4.4) in the control group. The mean total motile sperm count in the second sample was 3.52 x 10(6) (SD +/- 1.46 x 10(6); range 0.9-3.7) in the study group. The mean total motile sperm count in the final inseminate was 7.35 x 10(6) (SD +/- 1.90 x 10(6); range 2.9-10.6) in the study group. The difference in total motile sperm counts between the study and the control group was statistically significant (P < .001). There were 6 pregnancies in the study group, providing a pregnancy rate of 15.3%, whereas there were 5 pregnancies in the control group, representing a pregnancy rate of 10% (P = .44). We concluded that although it does not increase the pregnancy rate significantly, obtaining a second semen sample when the motile sperm yield of the first semen sample is 1 million to 5 million significantly increases the total motile sperm count in the final inseminate.     

Electroejaculation following retroperitoneal lymphadenectomy

Transrectal electroejaculation was performed in 24 men who were anejaculatory from retroperitoneal lymphadenectomy. Of the men 23 had undergone retroperitoneal lymphadenectomy because of testis cancer. Seminal emission was achieved in all patients. In 21 patients greater than 10 x 10(6) progressively motile sperm with normal morphology were obtained. The average sperm count and motilities obtained were 289 x 10(6) and 18%, respectively, for the antegrade fractions, and 2,051 x 10(6) and 13%, respectively, for the retrograde fractions. Of the 3 azoospermic failures 2 had chemotherapy-induced testicular damage and 1 had carcinoma in situ of the remaining testis. A total of 19 couples underwent artificial insemination with electroejaculated sperm. There were 7 pregnancies achieved in 74 insemination cycles (36.8% of the couples conceived for a 9% cycle fecundity). Routine semen parameters could not predict which couples would be successful in achieving pregnancy. There were 2 first trimester spontaneous abortions. Five healthy children have been born. Electroejaculation is an excellent treatment for anejaculation that persists following retroperitoneal lymph node dissection.     

Relationship between sperm motility assessed with the Hamilton-Thorn motility analyzer and fertilization rates in vitro

To determine which sperm movement characteristics are related to in vitro fertilization rates, semen and swim-up preparations used for in vitro fertilization in 108 patients were assessed using the Hamilton-Thorn HTM-2030 Motility Analyzer (HTMA) and other sperm tests. There were highly significant correlations between manual and HTMA results for sperm concentration (Spearman r = 0.881; P less than 0.001) and the percentage of motile spermatozoa (Spearman r = 0.580; P less than 0.001). The percentage of motile spermatozoa with average path velocities greater than 10 microns/s and greater than 20 microns/s, straight line and curvilinear velocity, linearity (straight line velocity vs curvilinear velocity), amplitude of lateral head displacement, and beat-cross frequency were significantly higher in the insemination medium after selection of motile spermatozoa by the swim-up technique than in the semen. Linearity (P less than 0.01), the percentage of morphologically normal spermatozoa (P less than 0.05) and straight line velocity (P less than 0.05) in semen, and the percentage of motile spermatozoa with average path velocities greater than 10 microns/s in both semen (P less than 0.05) and insemination medium (P less than 0.05) were significantly correlated with in vitro fertilization rate when examined by a nonparametric (Spearman) test. With logistic regression analysis of all data, only the diagnoses of male infertility and tubal disease, linearity in semen, and the percentage of motile spermatozoa with average path velocities between 10 and 20 microns/s in insemination medium were significantly related to in vitro fertilization rates.(ABSTRACT TRUNCATED AT 250 WORDS)     

Does transcervical intra-fallopian insemination improve pregnancy rates in cases of oligoteratoasthenozoospermia? A prospective,randomized study

The relatively low pregnancy rates (PR) after treatment of patients with oligoteratoasthenozoospermia (OTA) result in a search for different treatment modalities. The objective of this study was to assess the efficacy of transcervical intrafallopian insemination (IFI) with husband's semen in comparison to intrauterine insemination (IUI) in couples with OTA. A prospective, randomized study included 30 couples with OTA-related infertility (according to WHO criteria). The female patients underwent individually adjusted controlled ovarian stimulation by gonadotropins. Spermatozoa was prepared using the Percoll 70% technique and insemination was performed 36-40 h after human chorionic gonadotropin (HCG) administration. The Tomcat Catheter was used for IUI and the Jansen-Anderson Catheter for IFI to the fallopian tube leading to the ovary that contained more dominant follicles. The couples were divided according to sperm count, into group A (9 couples): < 10 mill ml-1 and group B (21 couples): > 10 mill ml-1. Within the groups the patients were randomly assigned for IUI or IFI treatment. Among group B couples, two pregnancies out of 15 IUI cycles (13.3% PR) and two pregnancies out of 18 IFI cycles (11.1% PR) were achieved. Group A patients completed 7 IUI and 9 IFI treatment cycles with no pregnancies observed. These data did not demonstrate a statistically significant advantage for either technique.     

Comparative study on density gradients and swim-up preparation techniques utilizing neat and cryopreserved spermatozoa

AIM: To 1) compare post-wash and post-thaw parameters of sperm processed with PureSperm density gradient technique and swim-up method; and 2) test the efficacy of two commonly available density gradient media PureSperm and ISolate. METHODS: This prospective study used semen specimens from 22 patients. Specimens from nine patients were processed by both PureSperm density gradient and swim-up method. These specimens were then cryopreserved. Thirteen specimens were processed by both PureSperm (40 % and 80 %) and Isolate (50 % and 90 %) double density gradient techniques. The two fractions processed by both PureSperm and swim-up were analyzed for post-wash sperm characteristics. Post-thaw analysis was done after 24 hours. Sperm fractions obtained after processing with PureSperm and ISolate were compared for post-wash sperm characteristics and ROS levels. Results: Specimens prepared with PureSperm had significantly higher median total motile sperm counts (TMSC) (32.2 x 10(6) vs. 17.6 x 10(6)), recovery rates (69.2 % vs. 50.0 %), and longevity at 4 hours (83.0 % vs. 55.0 %) compared to specimen prepared by swim-up. Post-thaw specimens also had a higher recovery and longevity at 4 hours with PureSperm as compared to the swim-up. Semen specimens processed by PureSperm had significantly higher total sperm count, TMSC, and percentage recovery rates (30.0 % vs. 19.7 %) than ISolate. CONCLUSION: Semen quality is better preserved in fresh and cryopreserved semen prepared with PureSperm density gradient compared to swim-up. A significant enrichment of sperm is observed with PureSperm compared to ISolate. Higher recovery rates of mature motile sperm obtained after PureSperm sperm preparation may be beneficial for successful ART.     

Intra-uterine insemination with prepared sperm vs. unprepared first split ejaculates. A randomized study

In this randomized prospective study, we determined the conception rate following intra-uterine insemination with washed and prepared sperm, or with the first portion of a split ejaculate, in couples with longstanding male (n = 27, 70 treatment cycles) or cervical infertility (n = 14, 29 treatment cycles). Folliculogenesis and ovulation were induced by human menopausal gonadotropin and human chorionic gonadotropin. Significantly more couples conceived in the male infertility group following intra-uterine insemination with washed sperm, than after intra-uterine insemination with split ejaculate (9 vs. 2; P less than 0.05), while no difference in pregnancy rate (2 vs. 2) was found by the two intra-uterine insemination methods in the cervical infertility group.     

Subnormal sperm parameters in conventional semen analysis are associated with discrepancies between fertilization and pregnancy rates in in-vitro fertilization and embryo transfer

Three hundred and twenty-eight consecutive treatment cycles in 168 couples were analysed retrospectively in order to examine the influence of conventional semen analysis results on the outcome of in-vitro fertilization and embryo transfer with respect to the occurrence of both fertilizations and pregnancies. All treatments were performed under maximally standardized and controlled conditions. Each of the three main determinants of the spermiogram, namely the concentration, motility and morphology of sperm in seminal plasma, was of significant importance for fertilization and subsequent pregnancy. Best correlations were achieved by counting the number of progressively (a+b) motile sperm and the number of normally formed sperm in seminal plasma. The pregnancy rate was reduced significantly in cases in which the sperm concentration was < 10 x 10(6) ml-1 (P < 0.01), or in which there was < 40% progressively motile sperm (P < 0.001), or < 30% normally formed sperm (P < 0.001). If more than one parameter in the spermiogram was abnormal, the fertilization rate depended mainly on the most disturbed sperm parameter. The implantation rate as well as the pregnancy rate was reduced significantly in patients with low progressive sperm motility and normal morphology rates. The difference could only be attributed partially to the lower number of embryos replaced. In conclusion, subnormal sperm quality seems to interfere with developmental stages beyond the process of fertilization.     

Comparison of sperm preparation methods: effect on chromatin and morphology recovery rates and their consequences on the clinical outcome after in vitro fertilization embryo transfer

The aim of this study was to evaluate the efficacy of swim-up, PureSperm gradient centrifugation and glass-wool filtration methods for semen preparation and to assess the possible enhancement of the quality of the subpopulation of spermatozoa in terms of sperm concentration, morphology and chromatin condensation. Moreover, to determine the effect of this semen processing technique on the clinical outcome after in vitro fertilization embryo transfer (IVF-ET). A total of 180 semen samples of patients' husbands who were undergoing IVF therapy were prepared by swim-up (G1, n = 60), PureSperm gradient centrifugation (G2, n=60) or glass-wool (G3, n=60) methods. Chromatin condensation was assessed by Chromomycin (CMA3), whereas sperm morphology was evaluated according to strict criteria. In all three semen processing methods, the percentage of chromatin condensed and morphologically normal spermatozoa was higher after semen processing in comparison with native semen samples. The proportion of normal chromatin condensed spermatozoa prepared in glass-wool filtration was significantly higher than that in swim-up (G.I, p=0.02) or PureSperm (G.II, p=0.001). In addition semen processing with PureSperm yields significantly a higher percentage of morphologically normal spermatozoa than swim-up (p < 0.001) or glass-wool method (p < 0.002). However, the fertilization, implantation and pregnancy rates, in turn were similar in all semen preparation methods. In conclusion, PureSperm gradient centrifugation yields a higher percentage of morphologically normal spermatozoa than shown in traditional swim-up or glass-wool filtration. However, the percentage of chromatin condensed spermatozoa was significantly higher after semen processing via glass-wool in comparison with the other two methods. Nevertheless, there were no significant difference in the fertilization, implantation and pregnancy rates of sperm prepared by means of swim-up, PureSperm or glass-wool filtration. Therefore, glass-wool filtration should be recommended as the first choice for semen preparation for Intracytoplasmic sperm injection (ICSI) technique as the natural selection is bypassed. Whereas, swim-up and PureSperm should be used for semen processing in IVF programme.     

影响供精人工授精成功率因素及最佳治疗方案探讨

目的：探讨影响供精人工授精(AID)成功率的因素及其最佳治疗方案。方法：98对夫妇行131个AID周期，其中8例行AID联合促排卵治疗3个周期以上未孕，遂改为ICSI。结果：AID共20例妊娠，且均发生在第1—3个周期，周期妊娠率为15．3％。女性患者年龄大于35岁和不孕年限大于10年的妊娠率显著降低。精子活动率在处理前妊娠组和非妊娠组相似，而处理后妊娠组显著高于非妊娠组(0．02＜P＜0．05)。每周期定时授精1次与2次的妊娠率无显著差异(P＞0．05)。8对夫妇行10个供精ICSI周期，有5例妊娠，周期妊娠率为50．0％。结论①无精子症患者的治疗首选供精AID；②AID的妊娠率随女性患者年龄的增长和不孕年限的延长而下降；③增加授精频率并不提高妊娠率；④当AID联合促性腺激素3个以上周期未孕时，ICSI将是最佳选择。     

Treatment of infertility due to anejaculation in the male with electroejaculation and intracytoplasmic sperm injection

PURPOSE: We tested the hypothesis that spinal cord injury and/or anejaculation affects the outcome of intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: From November 1993 to October 1998 we obtained and prospectively reviewed data on 34 ICSI cycles using electroejaculated sperm, 620 male factor infertility ICSI cycles using normal ejaculated sperm and 120 cases of obstructive azoospermia, in which microsurgical epididymal aspiration and testicular sperm extraction-ICSI were done in 93 and 27, respectively. RESULTS: A total of 34 ICSI cycles were performed in 17 couples with male infertility due to anejaculation secondary to spinal cord injury in 10 patients and retroperitoneal lymph node dissection in 5, and idiopathic in 2. In all 17 couples at least 3 previous intrauterine insemination cycles had failed. After electroejaculation 11 men had oligozoospermia and 6 normal sperm density. Median sperm retrieval volume plus or minus standard deviation was 1.9 +/- 1.9 ml., median sperm concentration 70.7 +/- 60.2 x 106 sperm per ml., median motility 10.7% +/- 10.8% and median forward progression 2.3 +/- 0.5 (scale 1 to 4). In the anejaculation group ICSI resulted in a median fertilization of 60% +/- 28%, 15% pregnancies per cycle and 29% pregnancies per couple. In the control group of 620 ICSI cycles from ejaculated specimens obtained from male patients with infertility median fertilization was 58% +/- 26%, and there were 39% pregnancies per cycle and 47% pregnancies per couple. The rate of pregnancies per embryo transfer and per couple was higher in the control than in the electroejaculation-ICSI group (p <0.05). However, there was no statistically significant difference in the fertilization rate. CONCLUSIONS: ICSI or in vitro fertilization is a viable alternative for patients with anejaculation in whom intrauterine insemination failed. While the fertilization rate is similar in these couples, the pregnancy rate is significantly lower than that achieved with ejaculated specimens from patients with severe male factor infertility. ICSI is a viable alternative for a patient with anejaculation in whom intrauterine insemination or in vitro fertilization failed.     

Computer-assisted image analysis of sperm concentration in human semen before and after swim-up separation: comparison with assessment by haemocytometer

Evaluation of male fertility is based predominantly on results from semen analysis and determination of the sperm concentration is one of the main parameters of the analysis. The availability of a fully automated videomicrographic digital image analyser would offer both an objective and rapid method for determination of the sperm concentration. In the present study the sperm concentration in 327 semen samples was determined by haemocytometer according to the World Health Organization guidelines, and also by a computer-assisted digital image analyser system. Results were classified according to the routine procedure (haemocytometer) before statistical analyses. The computerized measurements caused a shift to the right in the frequency distribution of sperm concentration. Sperm concentrations were more often overestimated significantly (P less than 0.001) by the computerized measurements in semen samples with concentrations up to 80.0 x 10(6)/ml. This overestimation seemed to be caused by the presence of particles in seminal plasma that were recognized incorrectly as sperm by the computer program. The computerized digital image analyser gave an average sperm concentration of 2.2 +/- 0.6 x 10(6)/ml (mean +/- SEM) in 17 azoospermic semen samples while the routine procedure did not detect the presence of sperm cells. After removing the seminal plasma by washing and centrifugation with culture medium, and using the swim-up procedure to harvest motile sperm, the computerized measurements showed comparable results with the routine procedure for those sperm preparations (n = 44) with sperm concentrations greater than 5.0 x 10(6)/ml.(ABSTRACT TRUNCATED AT 250 WORDS)     

The Split Ejaculate: Assessment of Fertility Potential Using Two in vitro Test Systems

Semen samples (split & whole ejaculates) were obtained from 12 normal men (group A) and 8 oligospermic infertile men with sperm concentrations of less than 20 x 10(6) sperm/ml (group B). All samples were evaluated by standard semen analysis, bovine cervical mucus penetration assay (CMPT), and, in all cases with sufficient sperm, in the human spermatozoa zona-free hamster in vitro penetration assay (SPA). In group A the motile sperm concentration was significantly higher in the ejaculated material of the first two contractions (fraction I or FI) than in the remainder of the ejaculate (fraction II or FII) (p less than 0.02). No significant differences were observed in sperm penetration into zona-free hamster ova or bovine cervical mucus by sperm from FI, FII or the whole ejaculate. Motile sperm concentration was significantly correlated with sperm penetration into bovine cervical mucus (r = 0.65, p less than 0.01), but not into zona-free hamster ova (r = 0.01 NS). In the samples collected by group B, the mean sperm concentration and motile sperm concentration were higher in the first (FI) than in the second (FII) fractions of the split ejaculate or the whole ejaculate (p less than 0.05). No significant differences were found among the FI, FII and the whole ejaculate semen samples for penetration of sperm into bovine cervical mucus. Sperm concentration and motile sperm concentration were significantly correlated with sperm penetration into bovine cervical mucus (r = 0.58, p less than 0.01 and r = 0.57, p less than 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of the effect of a cervical cap device on sperm functional characteristics in vitro

Intracervical insemination continues to be employed for homologous and donor insemination in natural and stimulated cycles. Efficacy studies for potential fertility involve in vivo assessment; however, in vitro testing of particular sperm function(s) critically involved in fertilization is an important component of such evaluation. We report here on the in vitro evaluation of the effects of the silicone Veos cervical cap (Veos, London, UK) on sperm function. Donor semen was exposed to the Veos cervical cap or a sterile 15-cc centrifuge tube (control), or treated with the spermicide nonoxynol-9 (5 mg x ml(-1) in saline) for 4 h at 37 degrees C and 5% CO2 in water-saturated air. After exposure, motility characteristics, both in semen and in spermatozoa processed by standard swim-up procedure, cervical mucus penetration and sperm-zona pellucida interaction using the hemizona assay were assessed. Results indicated that exposure to the Veos cervical cap had no effect on either sperm motility characteristics or sperm-zona pellucida interaction. A small but significant difference was observed for cervical mucus penetration (P = 0.05); however, for both the control and treated groups, vanguard spermatozoa exceeded manufacturer's guidelines for a normal test, a penetration distance of > or = 30 mm. As expected, nonoxynol-9 was a potent inhibitor of sperm function. Lack of adverse effects on in vitro spermatozoa functional characteristics after exposure to the silicone Veos cervical cap supports its addition to the repertoire of fertility treatment modalities when cervical insemination is indicated.