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PubMed2002

The effects of female age on the outcome of testicular sperm extraction and intracytoplasmic sperm injection in infertile patients with azoospermia.

INTRODUCTION: Testicular sperm extraction (TESE) is well-defined procedure for surgical sperm retrieval in obstructive and non-obstructive azoospermia. This study was focused on the effectiveness of testicular sperm extraction and intracytoplasmic sperm injection (ICSI) for azoospermic men with different female age subgroups. MATERIALS AND METHODS: A total of 107 men with azoospermia underwent TESE and ICSI treatment. The women were examined in three groups 20-29, 30-34 and 35 years or older. The main outcome in this study was fertilization and pregnancy rates with TESE and ICSI. RESULTS: Spermatozoa were successfully retrieved during 97 of 107 (90.7%) TESE attempts, resulting in the fertilization of 286 of 563 (50.4%) injected metaphase II oocytes. Two hundred and fifty-five of them were transferred (89.8%). The clinical pregnancy rate and ongoing pregnancy rate per embryo transfer were 22.5% and 20.6% respectively. When comparing the fertilization and pregnancy rates, it was observed that women between the ages of 20-29 years had significantly higher pregnancy rates than women over 34 years of age (p < 0.05). CONCLUSION: The female age is a major factor in determining successful implantation in ICSI.

Adult↗
PubMed1996

Electroejaculation and assisted reproductive techniques for anejaculatory infertility.

OBJECTIVE: To report our experience with electroejaculation combined with intrauterine inseminations (IUI) and other assisted reproductive technologies. METHODS: Anejaculatory men desirous of fertility were evaluated in our Assisted Reproductive Program. Between March 1991 and December 1994, 26 men (age 24-48) underwent a total of 84 electroejaculations. Causes of anejaculation included spinal cord injury (n = 23) and retroperitoneal lymph node dissection (n = 3). Female partners were super-ovulated to improve the pregnancy rate. The success in sperm retrieval and pregnancy rates were determined, and the quality of electroejaculates was evaluated. RESULTS: Seventy-seven of 84 (92%) electroejaculations were successful, defined by retrieval of more than 10 x 10(6) total and more than 10(6) motile spermatozoa. Mean sperm count was 65 million/mL (range 0-569), but mean motility was only 16% (range 0-66). Mean normal morphology was 27% (range 0-71). Ten couples attempted conception. Fifty cycles of IUIs were performed, resulting in four normal term infants and one spontaneous abortion (pregnancy rate 10% per IUI). One patient failed to conceive with eight cycles of IUIs but became pregnant with in vitro fertilization-embryo transfer with micromanipulation using electroejaculates; she delivered a set of healthy twins. Two couples elected donor sperm insemination after failing to conceive by IUI with electroejaculates; both became pregnant. CONCLUSION: Electroejaculation offers an encouraging pregnancy opportunity for anejaculatory men who otherwise are considered infertile. Marked asthenospermia is observed in electroejaculates, the etiology of which remains obscure. Further studies to elucidate the cause may improve pregnancy rates.

Adult↗
PubMed1994

Production of motile acrosome-reacted mouse sperm with nanomolar concentration of calcium ionophore A23187.

A method to generate a population of motile, acrosome-reacted mouse sperm is described. Sperm retrieved from the cauda epididymis and vas deferens were first capacitated in a 3% bovine serum albumin (BSA) containing medium. Sperm were then resuspended in medium with low BSA content (0.01%) and treated with 30 nM of the calcium ionophore, A23187, which was added as a single dose of 30 nM for 15 min at 37 degrees C; or three sequential 10 nM doses over three 5 min intervals. Approximately 55-60% of the treated sperm population became acrosome reacted. The motility of the treated sperm sample was 40-65%, slightly lower than that of the control sperm, following addition of medium containing 3% BSA. This is in contrast to the < 10% motility observed for capacitated mouse sperm treated with 10 microM 23187, a concentration that had been used by other investigators to induce the acrosome reaction. The ultrastructure of the 30 nM A23187-induced acrosome-reacted sperm was similar to that of the acrosome-reacted sperm induced by solubilized zonae pellucidae. These motile, acrosome-reacted sperm were able to penetrate zona-free mouse eggs at a higher rate than the control sperm. Thus this method of treatment will be useful for further physiological experimentation with acrosome-reacted sperm.

Acrosome↗
PubMed1978

Intrauterine insemination of isolated motile sperm.

Semen from infertile males was filtered through liquid albumin, and sperm retrieved from the most distal fraction were used for intrauterine insemination. Compared with the ejaculate, the isolated specimens contained fewer sperm but an increased percentage of motile sperm. The fractions were also free of the debris (white cells, agglutinated sperm, immature forms, and particulate matter) commonly seen in the ejaculates. Despite the improvement in motility none of the 19 women who were treated for a total of 67 cycles became pregnant. It is possible that even the best sperm from a poor specimen may have intrinsic defects that interfere with their ability to fertilize.

Female↗
PubMed2001

Intratesticular Doppler flow, seminal plasma nitrites/nitrates, and nonobstructive sperm extraction from patients with obstructive and nonobstructive azoospermia.

OBJECTIVE: To prospectively evaluate the role of intratesticular vascular flow in modulating sperm function in men with obstructive and nonobstructive azoospermia. The correlation of testicular Doppler values with nitric oxide and testicular sperm extraction was further evaluated. DESIGN: Prospective study. SETTING: Assisted reproduction unit at a university center. PATIENT(S): Twenty-eight men with azoospermia undergoing sperm extraction for intracytoplasmic sperm injection. INTERVENTION(S): Ultrasound and color Doppler scanning of the testes. Testicular sperm retrieval and nitrite/nitrate assay. MAIN OUTCOME MEASURE(S): Doppler analysis of testicular transmediastinal artery, plasma and seminal plasma nitrite/nitrate values, and sperm extraction histopathology. RESULT(S): The pulsatility index (PI) of the transmediastinal artery was higher in patients with nonobstructive azoospermia (PI = 1.40 +/- 0.13) than in those with obstructive azoospermia (PI = 1.09 +/- 0.15; P=.011). Seminal plasma nitrite/nitrate concentrations were more elevated in cases of obstructive azoospermia than in gonadal failure. Unsuccessful sperm recovery was observed in four patients who showed the worst indices of gonadal failure. In this subgroup, a transmediastinal PI value >1.50 was always observed. CONCLUSION(S): Doppler analysis of the transmediastinal artery and nitrite/nitrate seminal plasma concentrations are useful for distinguishing between obstructive and nonobstructive azoospermia and allow the identification of the presence of spermatozoa within the testes.

Adult↗
PubMed2002

[Testicular sperm extraction for non-obstructive azoospermia: results with conventional and microsurgical techniques].

Testicular sperm extraction (TESE) was performed on patients with non-obstructive azoospermia using the conventional or microdissection technique. First, conventional TESE was attempted on all patients. If the sperm was retrieved successfully, the TESE procedure was terminated. Microdissection TESE was indicated only for patients for whom the conventional sperm retrieval was unsuccessful. Sperm was successfully retrieved with conventional TESE from four (24%) of 17 patients with non-obstructive azoospermia. The remaining 13 patients underwent microdissection TESE and sperm was successfully retrieved from four of them. Application of the microdissection technique resulted in an improvement in sperm retrieval rates from 24% with conventional TESE to 48% when combined with the microdissection technique. None of the patients showed any acute or chronic complications after TESE. Intracytoplasmic sperm injection (ICSI) was performed on five of the eight patients with successful sperm retrieval and the partner of four of these patients became pregnant and progressed to delivery. These results indicate the usefulness of microdissection for improving the sperm retrieval rate in men with non-obstructive azoospermia.

Adult↗
PubMed2005

Computational models for prediction of IVF/ICSI outcomes with surgically retrieved spermatozoa.

IVF/intracytoplasmic sperm injection (ICSI) using surgically retrieved spermatozoa (SRS) is a key option in the treatment of severe male infertility. It was aimed to develop a computational model for the prediction of this modality's outcome. A dataset of 113 exemplars, derived from patients who underwent IVF/ICSI with SRS, was retrospectively analysed. The dataset, containing input features maternal age, sperm retrieval technique, type of spermatozoa used, type of male factor and output intrauterine pregnancy, was randomized into a modelling ('training') set of 83 and cross-validation ('test') set of 30. neUROn++, a set of C++ programs, was used to model the dataset using linear and quadratic discriminant function analysis, logistic regression, and neural computation. A 4-hidden node neural network was found to have the highest accuracy, with a test set receiver operator characteristic (ROC) curve area of 0.783. Reverse regression of this neural network showed maternal age to be the most significant feature in predicting pregnancy (P = 0.025), followed by sperm type (P = 0.076). Type of male factor (P = 0.47) and sperm retrieval technique (P = 0.88) did not predict outcome. In summary, a neural network of clinical relevance was found to be superior in terms of IVF/ICSI outcome prediction. Future media deployment is planned.

Adult↗
PubMed2002

Identification, localization and functional activity of oxytocin receptors in epididymis.

Oxytocin (OT) is a neurohypophysial hormone with unclear physiological functions in the male. Several previous studies indicated that OT might have a role in the ejaculatory process, stimulating sperm release from the epididymal storage. In this study we investigated on the presence and function of OT receptor (OTR) in rabbit and human epididymis. By using RT-PCR, Western and binding studies, we found that OTR gene and protein is expressed in the human epididymis and stimulates in vitro contractility. The immunolocalization of OTR suggests that the receptor is not only present in the smooth muscle cells of the human epididymis but also in the epithelial compartment. Experiments performed in rabbit epididymal epithelial (rEE) cells in culture indicate that OT induces the release of an other potent stimulator of epididymal contractility, endothelin-1 (ET-1), Blocking the ET(A) subtype of the ET-1 receptors, by using a specific antagonist (BQ-123), partially counteracts the contractile effect of OT, suggesting positive interactions between the two peptides in regulating epididymal contractility. Finally, to investigate whether an acute OT administration increases sperm release also in humans, we treated oligozoospermic patients with an intravenous bolus of OT (2.5 IU), just before sperm collection. In a small, single blind study, we found that OT almost doubled sperm retrieval when compared with vehicle administration. Our results indicate that OT might have physiological functions also in the male, controlling epididymal motility and sperm progression through the male genital tract.

Adult↗
PubMed2002

Cryopreservation, screening and storage of sperm the challenges for the twenty-first century.

The advent of HIV and the serious nature of the sequelae resulted in a major reassessment of artificial insemination practices in the UK. The development of human semen cryopreservation had enormous impact on reproductive medicine and the availability of cryopreserved quarantined donor semen became a mainstay for the treatment of male infertility in the UK. The regulation and accreditation of assisted reproductive technologies and the introduction of peer-reviewed guidelines have largely standardized clinical and laboratory practice. The introduction of assisted fertilization techniques such as intracytoplasmic sperm injection, testicular sperm retrieval and improved oncology treatments have placed pressure on reproductive biologists and cryobiologists to design and use cryopreservation protocols for the optimum survival of sperm.

Cryopreservation↗
PubMed1999

Role of the urological surgeon in the age of intracytoplasmic sperm injection.

In the last few years, it has been demonstrated that intracytoplasmic sperm injection is an effective procedure for treating patients with severe male infertility. Before this technique was available, the urological surgeon involved in the reproduction field dealt mainly with reconstructive (vasoepididimostomy, vasovasostomy) or endoscopic surgery of the seminal duct as well as varicocelectomy. Nowadays, the urological surgeon continues to participate in the field of reproductive surgery by applying methodologies which enable natural conception, and has to be involved in all aspects of sperm retrieval techniques.

Ejaculatory Ducts↗
PubMed1995

Practical evolution and application of direct intracytoplasmic sperm injection for male factor and idiopathic fertilization failure infertilities.

OBJECTIVE: To analyze the introduction of a new assisted fertilization technique for the treatment of severe male factor and idiopathic fertilization failure infertilities. DESIGN: Retrospective analysis of 16-month clinical application of IVF-ET where insemination was performed solely by direct intracytoplasmic sperm injection. SETTING: Clinical IVF-ET program. PATIENTS: Ninety-two couples undergoing 105 cycles of sperm injection. RESULTS: One hundred embryo transfers yielded 28 viable pregnancies (28%) from which eight normal deliveries have occurred to date. Complete cleavage arrest or fertilization failure occurred in four cycles, and one couple had all embryos cryopreserved. One thousand one hundred forty-three eggs were injected of which 173 (15%) degenerated. Four hundred seventy-nine of the surviving 970 eggs became normally fertilized (49%), and 381 of these zygotes (79.5%) developed suitably for cryopreservation or for transfer. Thirty-four of 310 embryos transferred implanted, yielding an implantation rate of 11%. Both testicular and epididymal sperm were used successfully to achieve fertilization and pregnancies, as was sperm retrieved by electroejaculation. Older women and couples suffering from prior idiopathic fertilization failure had a markedly poorer outcome. CONCLUSIONS: These results confirm that the intracytoplasmic sperm injection technique is a successful form of assisted fertilization that can be applied to a wide range of couples at significant risk from fertilization failure.

Adult↗
PubMed2001

Percutaneous epididymal sperm aspiration followed by intrauterine insemination--a new approach to achieve a pregnancy in infertility due to obstructive azoospermia.

Standard approach towards treatment of obstructive azoospermia whether congenital or acquired is mostly microsurgical epididymal sperm aspiration (MESA) and to some extent percutaneous epididymal sperm aspiration (PESA) both being followed by intracytoplasmic sperm injection (ICSI). However in the present series for sperm retrieval in case of obstructive azoospermia the author conducted a series of 55 cases of PESA along with only 4 cases of MESA. The epididymal sperm aspiration (ESA) was followed by intrauterine insemination (IUI) (whenever required quantity of motile sperm was collected) in place of ICSI with full consent of the couples. With the help of this ESA-IUI programme it was possible to achieve reasonable success rate viz, 20% pregnancy rate (PR) per cycle for PESA-IUI and 14.28% for MESA-IUI.

Adult↗
PubMed1997

Testicular sperm extraction with intracytoplasmic sperm injection for nonobstructive azoospermia.

OBJECTIVES: To provide fertility for men with nonobstructive azoospermia. METHODS: A retrospective review of treatment results at a university infertility center was undertaken. Sixteen couples entered an attempted in vitro fertilization (IVF)-intracytoplasmic sperm injection (ICSI) cycle for treatment of nonobstructive azoospermia. Each man was azoospermic, and the male factor diagnosis of nonobstructive azoospermia was made on testis biopsy for 14 men and on clinical grounds for 2 men. Sperm were retrieved by testicular biopsy on the day of oocyte retrieval. Results of testicular examinations, serum follicle-stimulating hormone levels, and testicular histology as well as evaluation of the success rates of sperm retrieval, fertilizations, and pregnancies were made. RESULTS: Sperm were extracted from testis biopsies in 10 of 16 (62%) testicular sperm extraction (TESE) attempts. For cycles in which sperm were retrieved, normal fertilizations were achieved for 51 of 98 (52%) mature oocytes injected with testicular sperm in 10 couples. Biochemical pregnancies were achieved for 6 of 16 (38%) couples, with clinical pregnancies during 5 of 16 (31%) attempts at sperm retrieval, and ongoing pregnancy and subsequent live delivery for 4 of 16 (25%) attempts. CONCLUSIONS; Pretreatment clinical parameters are unable to predict which men with nonobstructive azoospermia will have spermatozoa retrieved by TESE. When sperm are found, clinical pregnancies can occur for half (5/10) of these couples using TESE with ICSI, with ongoing pregnancy and delivery for 4 of 10 (40%). Many men with nonobstructive azoospermia will have retrievable sperm with testis biopsy that are suitable for ICSI; however, 6 of 16 (38%) couples will not have sperm retrieved with TESE and may undergo an unnecessary IVF procedure.

Adult↗
PubMed1992

Epididymal sperm in assisted reproductive technology.

Infertile men with obstructive azoospermia mainly due to congenital absence of the vas deferens (CAVD) now have the option of trying to father their own progeny. In fact, in the last five years epididymal sperm retrieval microsurgically have been successfully used for in vitro fertilisation of human oocytes. In this report the clinical results of 98 consecutive procedures of microsurgical epididymal sperm aspiration (MESA) combined with in vitro fertilisation (IVF) and tubal embryo transfer (TET) are described. An overall fertilisation rate of 17% and a pregnancy rate of 36% per transfer is reported. Five of the 18 pregnancies resulted in abortion (27%) and 13 were delivered at term. Additionally, extra embryos for freezing and potential use for future attempts were made available for 13 couples. Men with CAVD have also allowed the study of spermatogenesis, immunological response and sperm disposal mechanisms in condition of chronic obstruction.

Adult↗
PubMed1999

[Microsurgical epididymal sperm aspiration (MESA), testicular biopsy and intracytoplasmic sperm injection (ICSI) in the treatment of male infertility].

OBJECTIVE: To assess if ICSI using epididymal or testicular spermatozoa is effective in the treatment of couples with male factor infertility. MATERIAL AND METHODS: 56 couples suffering from male infertility underwent a total of 88 treatment cycles of ICSI, in combination with microsurgical epididymal sperm aspiration (MESA): 62 cycles, or testicular sperm retrieval: 26 cycles. RESULTS: Overall, fertilization rate was 63% per injected metaphase II oocyte. In MESA group, fertilization rate was 64% compared to 61% embryos in testicular sperm aspiration. The overall pregnancy rate was 26% per started ICSI cycle. Pregnancy rates were also similar in both group: 24% for MESA and 32% for testicular sperm extraction. 9 pregnancy (41%) were obtained using cryopreserved epididymal sperm (6 cases) or testicular sperm (3 cases). CONCLUSION: ICSI combined with epididymal or testicular sperm achieved a high fertilization and pregnancy rate. It constitutes an efficient alternative in the treatment of men suffering of testicular failure or azoospermia not amenable to surgical reconstruction.

Adult↗
PubMed2006

Morphological and functional characteristics of chilled semen obtained from domestic feline epididymides (Felis catus).

The objective of this study was to determine morphological and functional characteristics of semen retrieved from the feline epididymis before and after cooling. Sixteen adult male cats were orchiectomized. The distal portion of the epididymis and proximal part of the deferent ducts were dissected and squeezed to obtain their content. After centrifugation, the supernatant was removed, sperm were resuspended in a 0.9 mL Tris-fructose-citric acid extender containing 20% egg yolk, aliquoted into three 0.3 mL samples, placed in a refrigerator (4.8 degrees C) and cooled (0.5 degrees C/min). Semen evaluations were performed on four occassions: immediately after epididymal sperm retrieval (T0), and at 24 h (T1), 48 h (T2) and 72 h (T3) after cooling. On each occasion, progressive motility, vigor and sperm morphology were determined. Mean motility and vigor decreased (P < 0.05) between each successive examination. Although the majority of sperm cell damage occurred within the first 24 h, there was a decrease (P < 0.05) in mean percentage of morphologically normal sperm between T0 and each evaluated time (T1, T2, T3) after cooling, due to an increase in coiled and bent sperm tails. Further studies are needed to evaluate the effects of cooling on the fertilizing capacity of cat epididymal spermatozoa in assisted reproduction programs.

Animals↗
PubMed2000

Intracytoplasmic sperm injection with cryopreserved testicular spermatozoa.

To assess if testicular sperm cryopreservation is a valid alternative to repetition of testicular sperm retrieval techniques, results of a cryopreservation technique in pills have been retrospectively analyzed. Enough motile spermatozoa for ICSI were obtained in 172 from 190 (90.5%) frozen-thawed testicular sperm samples. Overall, 249 couples underwent 390 ICSI cycles, 156 using fresh and 234 using cryopreserved testicular sperm. Mean two-pronuclear fertilization rates per cycle were not significantly different after ICSI with fresh (62.0%) or with cryopreserved (63.2%) spermatozoa. Mean embryo cleavage rate per cycle was higher in the fresh (90.6%) than in the cryopreserved (84.6%) group (P = 0.016). However, clinical pregnancy rates per cycle (28.2% with fresh vs 27.8% with cryopreserved), implantation rates (12.2% vs 13.1%) and ongoing pregnancy rates per cycle (22.4% vs 21.8%) were not significantly different. Cryopreservation of testicular spermatozoa is an effective technique that can be used both in obstructive and in non-obstructive azoospermia.

Adolescent↗
PubMed2004

Exposure to trichloroethylene and its metabolites causes impairment of sperm fertilizing ability in mice.

Trichloroethylene (TCE) is a prevalent occupational and environmental contaminant that has been reported to cause a variety of toxic effects. Here, we have undertaken studies to test the hypothesis that TCE exposure adversely affects sperm function and fertilization. Sperm retrieved from mice exposed to TCE (1000 ppm) by inhalation for 1 to 6 weeks were incubated in vitro with eggs isolated from superovulated female mice. The number of sperm bound per egg was significantly decreased when mice were exposed to TCE for 2 and 6 weeks but not at exposures of 1 and 4 weeks. In vivo fertilization was also determined in superovulated female mice mated with males exposed to TCE for 2 to 6 weeks. The percentages of eggs fertilized, as assessed by the presence of two pronuclei, were significantly decreased after 2 and 6 weeks of TCE exposure. A slight but insignificant decrease was observed after 4 weeks of TCE exposure. The direct effects of TCE and its metabolites, chloral hydrate (CH) and trichloroethanol (TCOH), on in vitro sperm-egg binding were also investigated. Sperm-egg binding was significantly decreased when sperm were pretreated with CH (0.1-10 microg/mL). Significantly lower levels of sperm-egg binding were also detected with TCOH (0.1-10 microg/mL), although the decreases were not as pronounced as those for CH. These results showed that TCE exposure leads to impairment of sperm fertilizing ability, which may be attributed to TCE metabolites, CH, and TCOH.

Acrosome Reaction↗