Prognostic value of objective semen parameters in an in vitro fertilization program

Purpose: The basic semen parameters seem to have a limited predictive value in male fertility. Could other objective sperm analyses be helpful in the choice of the most adapted assisted procreation technique?Methods: This study concerns 78 infertile couples with insemination failures. For each semen, 21 objective parameters are analyzed in fresh semen and after sperm selection procedure. The 78 couples are then included in an IVF protocol and classified into two groups: fertile (at least one cleaved embryo is obtained) and infertile.Results: Using multiple variant discriminant factorial analysis, we have found nine nonconventional parameters which induce us to define two classes of semen. These two classes fit with the classification into fertile and infertile groups in 74.4% of the cases.Conclusions: So these parameters allow us to predict the chance of obtaining embryos during an IVF trial and to choose for each couple the most appropriate technique: IVF or ICSI.Key words: acrosome reaction, hyperactivated motility, in vitro fertilization, intracytoplasmic sperm injection, sperm kinetics     

Identification of male-factor semen samples prior to insemination and in vitro fertilization

Semen analyses carried out as part of the clinical in vitro fertilization or intrauterine insemination protocols provide important information that determine the type of clinical treatment of the male partner and the sperm processing method. It is postulated that the sperm of male-factor patients cannot survive hypoosmotic stress conditions because of defective sperm membrane function. To test this, 0.1 ml of semen from each of 102 patients was placed in 1.0 ml of 150 mosmolliter eosin citrate fructose solution and incubated for 30 min at 37°C. The percentage viability of the sperm cells was then determined. The results indicated that patients with two or more abnormal semen parameters had a significantly lower percentage viability while in the hypoosmotic solution (40.6±4.7%), in contrast to non-male-factor patients (69.0±1.6%). Donor sperm (N=32) serving as controls (73.3±2.1%) had a viability in hypoosmotic solution similar to that of non-male-factor patients. The data suggest that sperm of male-factor patients are less able to survive the hypoosmotic stress conditions as shown by the percentage viability in hypoosmotic solution and emphasize the impertance of using less stressful sperm processing methods for in vitro fertilization or insemination in these patients.Presented in part at the 46th Annual Meeting of the American Fertility Society, Washington, D.C., 1990.     

The relationship of semen parameters to fertilization in patients participating in a program of in vitro fertilization

Approximately 80% of the patients in a program of in vitro fertilization (IVF) will fertilize an oocyte. The purpose of this study was to determine which parameters of the semen analysis influence fertilization in vitro. Of 120 patients participating in an in vitro fertilization program, 98 achieved fertilization of at least one mature oocyte and 22 did not. Ovulation induction was standardized and patients whose sperm was exposed to at least one mature oocyte (by light microscopy) were included in the study. Semen washing was accomplished using a swim-up technique. Semen parameters were assessed both before (raw) and after washing. Following insemination with 100,000 motile sperm, fertilization was determined by the presence of pronuclei or cleavage. Mean sperm count and motility were higher in patients who fertilized. However, morphology was similar. Fertilization was more likely to occur with a raw density>104 million/ml and a motility>64%, as well as with a density>18 million/ml and a motilkity>86% following washing. Furthermore, washing lowered sperm counts by 75% and increased motility by 25% but had no effect on morphology. This study demonstrates that sperm count and motility, but not morphology, influence fertilization in a program of in vitro fertilization and that patients with higher counts and motility have and greater probability of fertilization.Presented at the Third Annual Congress of Andrology, April 27–May 2, 1985, Boston, Massachusetts.     

Successful use of pentoxifylline in male-factor infertility and previous failure of in vitro fertilization: A prospective randomized study

Objective Our objective was to determine whether the use of pentoxifylline (PF) would improve the in vitro fertilization (IVF) rate and outcome in couples with male factor infertility and previous failure of fertilization in vitro.Design This prospective randomized controlled study was conducted in an assisted conception unit.Materials and Methods Forty-nine couples with previous failed fertilization in vitro attributable to male factor or male-factor infertility without previous IVF were recruited for the study. Controlled ovarian hyperstimulation was performed using a combination of gonadotropin releasing hormone agonist and human menopausal gonadotropin. Oocytes of the same grade and maturity were inseminated with spermatozoa treated with PF or control spermatozoa. A maximum of three embryos was replaced after 48 hr and all other embryos were cryopreserved. Pregnancy outcome was followed up and evidence of fetal or neonatal anomalies reported.Results A significantly higher fertilization rate occurred in the group where oocytes were inseminated with spermatozoa treated with PF compared with controls (56.3 versus 30.7%; P<0.05). Fertilization occurred in 45 of the 49 cycles (92%). In seven cycles, only the oocytes that were inseminated with spermatozoa treated with PF fertilized, in contrast to only one cycle where the oocytes inseminated with control sperm fertilized (P<0.05). Fifty-seven PF and 31 control embryos were replaced and 11 clinical pregnancies occurred. Three of the pregnancies occurred in the seven cycles in which only PF embryos were replaced, one in the single cycle where control embryos were replaced and seven from the 37 cycles in which both PF and control embryos were replaced. There was no evidence of congenital malformations in any of the offsprings resulting from this study.Conclusion This study suggests that PF improves the fertilization rate and outcome in couples with male factor infertility and poor fertilization rates. This study does not suggest any increase in teratogenicity or evidence of congenital malformations in pregnancies following IVF cycles where PF was used.     

The efficiency of cryopreserved semen versus fresh semen for in vitro fertilization/embryo transfer

The efficiency of cryopreserved donor semen versus fresh donor semen in an in vitro fertilization/embryo transfer programme was evaluated. Thirty-nine in vitro fertilization/embryo transfer cycles were performed using fresh donor semen (group A) and 74 cycles were carried out using cryopreserved semen (group B). All patients anderwent a uniform controlled ovarian hyperstimulation using high doses of human menopausal gonadotropins. Oocytes were retrieved transvaginally under ultrasound imaging. Semen (fresh or frozen-thawed) were prepared for insemination by a washing technique. Each ovum was inseminated with 500,000–600,000 motile spermatozoa. No significant difference was noted between the two groups regarding female age, duration of infertility, and number of ova retrieved per aspiration. Even though the fertilization rate in group B was significantly lower than in group A (55.5±3.8 vs 70.4±3.5,P=0,008); pregnancy rates per embryo transfer were similar-39.3 and 38.5%, respecitively.     

Successful in vitro fertilization and pregnancy by micromanipulation with epididymal sperm

Purpose To examine the effectiveness of micromanipulation on the treatment of the patients with obstructive azoospermia, subzonal insemination or partial zona dissection was performed using epididymal sperm of the 23 patients, and the in vitro fertilization and embryo transfer (IVF-ET) outcomes were analyzed.Results Fertilized oocytes were obtained in 11 cycles of the 25 treated cycles, and six pregnancies were established by the subsequent ET. Through the analysis of 13 cycles in which both the micromanipulation and usual IVF insemination were performed, the fertilization rate per oocyte with the micromanipulation [25% (28/113)] was significantly higher than that with the usual insemination method [4% (4/102)]. The micromanipulation was most useful for the treatment of the patients with congenital absence of the vas deferens. Moreover, there was no need to aspirate a large volume of the epididymal fluid, but it was essential to prepare more than 10 mature oocytes retrieved.Conclusions These results indicate that micromanipulation with epididymal sperm is an effective treatment for obstructive azoospermia, and careful preparations of sperm and wellcontrolled ovarian hyperstimulation are necessary for successful IVF-ET.     

Intracytoplasmic sperm injection and conventional in vitro fertilization for sibling oocytes in cases of unexplained infertility and borderline semen

Purpose: In a prospective study, conventional IVF and intracytoplasmic sperm injection (ICSI) were performed on sibling oocytes of 22 patients with unexplained infertility (Group A) and 24 patients with borderline semen (Group B).Results: In Group A, there was no significant difference (P=0.070) in the fertilization rate per oocyte between ICSI (63%) and conventional IVF (50.7%), however, there was total failure of fertilization in conventional IVF in 5 of the 22 patients with IVF and none in ICSI. In group B, there was a significant difference (P<0.001) between the fertilization rate per oocyte in ICSI (59%) and conventional IVF (27.1%). There was total failure of fertilization in 11 patients after conventional IVF and none after ICSI.Conclusions: The study showed that 22.7% of unexplained infertility and 45.8% of patients with borderline semen would have lost their chance of embryo transfer completely because of total failure of fertilization if ICSI was not performed on some oocytes in this cycle.Presented in part at the IXth World Congress on In Vitro Fertilization and Alternate Assisted Reproduction, Vienna, Austria, April 3–7, 1995.     

Human pregnancy by in vitro fertilization (IVF) using sperm aspirated from the epididymis

Spermatozoa were collected by microaspiration from the corpus epididymidis of a 42-year-old man with secondary obstructive azoospermia and used for in vitro fertilization. At insemination 61% of the spermatozoa were motile, with a motility index of 157. One of five eggs was fertilized and this was subsequently transferred to the patient's wife at the two-cell stage. Ultrasound examination and changing hormone levels confirmed an ongoing pregnancy, which is currently at 30 weeks of gestation. This technique will provide a useful alternative for the management of some infertile men with obstructive azoospermia.     

Antisperm antibodies and in vitro fertilization

The purpose of this study was to investigate the influence of antisperm antibodies in the male, the female, or both partners on the outcome of in vitro fertilization treatment. The results in terms of ongoing pregnancies in the male and female antibody-positive group were the same as in the antibody-negative group. In the double antibody-positive group two of the three patients became pregnant. When high levels of antisperm antibodies were present on the spermatozoa, the fertilization rate was significantly reduced. In the female positive group no clear relationship between the antibody titer and the fertilization percentage could be detected. Abnormal semen quality was responsible for a much lower fertilization rate than the presence of antibodies. The conclusion of this study is that in vitro fertilization provides an equal chance of conception in couples with antisperm antibodies in comparison with couples with no antibodies if the other semen parameters are normal.     

Intracytoplasmic sperm injection overcomes previous fertilization failure with conventional in vitro fertilization

Objective: To evaluate the outcome of intracytoplasmic sperm injection (ICSI) in patients with previous idiopathic fertilization failure (≤20% fertilization rate) after conventional IVF.

Design: Retrospective analysis.

Setting: IVF program at a university medical center.

Patient(s): Twenty-five patients who underwent 38 ICSI cycles after experiencing unexplained fertilization failure with conventional IVF (group A) and 87 patients who underwent 118 ICSI cycles for male factor indications during the same period (group B).

Intervention(s): Intracytoplasmic sperm injection was performed in a subsequent cycle after fertilization failure with conventional IVF.

Main Outcome Measure(s): Outcomes of IVF were compared between groups A and B.

Result(s): Fertilization was achieved with ICSI in all patients with previous fertilization failure. The mean (±SD) fertilization rate (68% ± 21% vs. 64% ± 22%), implantation rate per embryo (22.6% vs. 20%), and delivery rate per cycle (47.3% vs. 49.1%) did not differ significantly between groups A and B. Overall, 72% of patients with previous unexplained fertilization failure had a successful pregnancy after ICSI.

Conclusion(s): Intracytoplasmic sperm injection can overcome unexplained fertilization failure caused by a potentially occult gamete abnormality, with the same fertilization, implantation, and pregnancy rates as are seen in patients with abnormal sperm parameters.     

Use of the total motile sperm count to predict total fertilization failure in in vitro fertilization

OBJECTIVE: To evaluate the capacity of baseline characteristics and total motile sperm count (TMC) to predict total fertilization failure (TFF) in patients undergoing IVF. DESIGN: Retrospective cohort study. SETTING: University hospital. PATIENT(S): Eight hundred ninety-two couples with a total of 1,569 consecutive IVF cycles. INTERVENTION(S): Prewash and postwash TMC during fertility workup and at the time of ovum pickup (OPU). MAIN OUTCOME MEASURE(S): Analysis of logistic regression and the receiver operating characteristic curve were used to determine which variables could be used to predict TFF. RESULT(S): The area under the curve (AUC) for prewash TMC during fertility workup was 0.72, similar to a combination of pre- and postwash TMC. At the time of OPU, both pre- and postwash TMC had an AUC of 0.73. A model based on selected baseline characteristics (male age, number of IVF cycles, indication for IVF, and prewash TMC during fertility workup) had an AUC of 0.75. A model at the time of OPU, including the number of oocytes, had an AUC of 0.80. CONCLUSION(S): The use of both models, one before start of the IVF cycle and one at the time of OPU, allows an accurate prediction of the chance of TFF and is useful in counseling patients on whether to opt for IVF or ICSI.     

Results using medium supplemented with either fresh or frozen stored serum in human in vitro fertilization

This study retrospectively compared the success of in vitrofertilization (IVF) among patients whose gametes had been incubated either in medium supplemented with freshly prepared pooled serum (331 cases) or in pooled serum which had been stored at –20°C prior to use (728 cases). Frozen stored serum was as effective as fresh serum with regard to the proportion of oocytes which fertilized and embryos which implanted and was not associated with any increased incidence of fetal loss during postimplantation development.     

Cumulative live birth rate following in vitro fertilization: Study of 5310 cycles

Most previous studies that calculated cumulative delivery rates following in vitro fertilization (IVF) treatments were limited in the number of cycles and the implementation of intracytoplasmic sperm injection (ICSI). Therefore, we assessed the yield of high-order consecutive IVF treatments (up to 14 consecutive cycles) with and without ICSI. Data from IVF cycles performed in a single center were retrieved from a computerized database. A total of 5310 cycles among 1928 patients were evaluated and cumulative delivery rates until the first delivery were calculated using life-table analyses. There were 1126 pregnancies resulting in 689 live births. Cumulative delivery rates reached 87% following 14 consecutive cycles. Cumulative delivery rates were higher following ICSI compared with cycles without (92.7% vs. 85.4%). In conclusion, each treatment cycle increased the cumulative delivery rate, resulting in a rate of 87% after 14 consecutive cycles. The introduction of ICSI resulted in the highest cumulative rates.     

Ovarian pregnancy after in vitro fertilization

We report a rare ovarian pregnancy after in vitro fertilization and transcervical embryo transfer. Ultrasound guidance of embryo transfer does not eliminate the risk of this potentially lethal complication.     

Successful fertilization,embryo development,and pregnancy in human in vitro fertilization (IVF) using a chemically defined culture medium containing no protein

A randomized control trial involving the fertilization and culture of human embryos in culture medium (T6) conraining either 10% maternal serum or no protein or amino acid supplement was carried out to assess the effect of deletion from culture of all fixed nitrogen on fertilization, embryo development, and embryo viability. There was no difference in fertilization rates (68 vs 69%), development of apparently normal embryos (96 vs 97%), pregnancy rate (18 vs 14%), or birth rate (13 vs 11%) between protein-containing and protein-free media. Deletion of protein from the culture medium may enable the constitution of more appropriate and defined culture media for human in vitro fertilization (IVF).     

Twin pregnancy following IVF treatment using frozen-thawed retrograde ejaculated sperm

Freezing retrograde ejaculated semen retrieved from a non-ilkalinized urine sample enabled the successful outcome of IVF treatment in a patient with mild congenital spina bifida.     

Male-factor infertility and in vitro fertilization

In vitro fertilization (IVF) was developed primarily as a treatment for female and idiopathic infertility. However, with the discovery that relatively few sperm are required to achieve fertilization in vitro, it was proposed that IVF could be used also as an effective treatment for male-factor infertility. This review deals with the work that has been carried out by various groups in this area of malefactor infertility. As the standards of classification and the presentation of results vary from group to group, this also shows that there is a need for some standardization of how patient selection and the presentation of results are carried out in the area of male-factor infertility.     

Predictive value of sperm morphology and movement characteristics in the outcome of in vitro fertilization of human oocytes

One hundred fourteen semen samples from Chinese males were analyzed for routine semen parameters including the semen volume, sperm count, percentage motility, and percentage normal morphology. Of these 114 samples, 54 also had movement characteristics of seminal and swim-up sperm evaluated by the computer image analyzer system (Cellsoft; Cryo Resources Co., New York). All semen samples were subjected to the swim-up procedure to harvest the motile sperm before inseminations of human oocytes. Fertilization was considered to have occurred when at least one oocyte was observed with two or more pronuclei. Semen samples were classified as infertile (0% fertilization rate;N=32) or fertile (>0% fertilization rate;N=82) before statistical analyses. There was a significant difference (P<0.005) in percentage normal morphology of seminal sperm between the fertile (mean±SE; 67.3±1.2%) and the infertile (59.3±2.2%) samples. The percentage normal morphology of seminal sperm correlated (r=0.3049;P<0.002) with the fertilization rate and this parameter was selected by the multivariate stepwise discriminant analysis as the discriminator capable of predicting the fertilization rate with 57.9% accuracy. Statistical analyses of samples where sperm movement was also evaluated demonstrated that there was significant differences (P<0.01) between the fertile (N=38) and the infertile (N=16) samples in percentage normal morphology of seminal sperm (67.8±1.8% vs 56.2±2.6%) and curvilinear velocity of swim-up sperm (89.2±3.5 vs 68.2±7.2 m/sec). The fertilization rates correlated with the percentage normal morphology of seminal sperm (r=0.3868;P<0.005) and velocity of swim-up sperm (r=0.3842;P<0.005). Multivariate stepwise discriminant analysis demonstrated that these two sperm parameters in combination were capable of predicting the fertilization rate with 74.1% accuracy. Our results indicate that seminal sperm morphology, coupled with computerassisted image analysis of movement characteristics of swim-up sperm, can help to predict the outcome of in vitro fertilization of human oocytes.     

Relation between antisperm antibodies and the rate of fertilization of human oocytes in vitro

To clarify further the role of antisperm antibodies in in vitro fertilization, the occurrence of antisperm antibodies on ejaculated sperm and in sera was determined by the immunobead binding assay in 67 couples after an unsuccessful in vitro fertilization cycle. Antisperm antibodies in maternal sera were associated with a failure of oocyte fertilization (P <0.02) or with fertilization of only 9–19% of the oocytes (P <0.01) in vitro. Antisperm antibodies were detected in sera from 13 of 24 women (54.2%) where no fertilization occurred, 9 of 14 women (64.3%) where less than 20% of the oocytes fertilized, and 3 of 19 women (15.8%) where greater than 40% of the oocytes fertilized. Antisperm antibodies in these sera were mostly IgG and directed against the sperm tail. Antibodies on the surface of ejaculated motile sperm were also associated with a low (9–19%) fertilization rate (P <0.01). Sperm-bound antibodies were detected in 2 of 24 men (8.3%) where no fertilization occurred, 5 of 14 men (35.7%) where less than 20% of the oocytes fertilized, and 0 of 19 men where fertilization was greater than 40%. Sperm-bound antibodies were mainly IgA and were tail-directed. Antisperm antibodies in sera of males were not related to the rate of fertilization. Antisperm antibodies were detected in female partners of 21 of 46 couples (45.7%) with unexplained infertility, 2 of 12 women (16.7%) with blocked tubes, 4 of 7 women (57.1%) with endometriosis, and 0 of 2 women with adrenal hyperplasia. There was no relation between the fertilization rate and the maternal age, number of oocytes harvested, or semen quality. We conclude that antisperm antibodies are present in sera from a high percentage of women with unexplained infertility and that antibodies reacting with sperm tails may directly interfere with fertilization in vitro or may be a surrogate marker for another factor that interferes with this event.