Multiple Testicular Sperm Extraction Research Articles

Endocrine Tests and/or Testicular Volume are Not Predictive of Successful Sperm Retrieval by Conventional Multiple Testicular Sperm Extraction in Nonobstructive Azoospermia

Objectives: This study aimed to determine the role of presurgical markers in the prediction of sperm retrieval by conventional Multiple Testicular Sperm Extraction in infertile Vietnamese men with nonobstructive azoospermia (NOA). Patients and Methods: Retrospective descriptive analysis of 136 infertile men with azoospermia, examined from August 2014 to July 2018. Patients underwent stepwise surgical sperm retrieval via percutaneous epididymal sperm aspiration, testicular sperm aspiration, then conventional multiple testicular sperm extraction in up to three locations, and procedures stopped as soon as sperm were detected. Factors were analyzed to determine the prediction of the likelihood of successful sperm retrieval, in men with NOA. Results: The overall success rate of sperm retrieval in men with azoospermia was 49.3%, but it was only 18.4% in NOA group. The difference in testicular volume between men with successful sperm retrieval and unsuccessful sperm retrieval was not statistically significant in NOA group (5.68 ± 2.37 vs. 4.46 ± 2.83, p = 0.138). The differences in the endocrine tests between the two groups were also not significant in terms of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (p [Formula: see text] 0.05). Multivariable analysis of predictive factors of sperm retrieval in NOA groups found no significant difference, except testicular density (p = 0.015). Conclusions: In infertile men with NOA, neither an endocrine test nor testicular volume should be used for predicting the results of surgical sperm retrieval by conventional multiple testicular sperm extraction.

445 Relevance of testicular histopathology on prediction of sperm retrieval rates after a conventional bilateral multiple TESE in case of non-obstructive azoospermia (NOA) and obstructive-azoospermia (OA)

To establish the relevance of testicular histopathology (TH) on sperm retrieval (SR) after testicular sperm extraction (TESE) in patients with non-obstructive azoospermia (NOA) and in obstructive azoospermia (OA) patients, already underwent a previous failure testicular fine needle aspiration (TEFNA). We evaluated a total of 82 azoospermic men, underwent TESE, referring to our Centre between January 2008 and March 2017. A general and genital physical examination, scrotal and trans-rectal ultrasound, semen analysis, hormone measurements, including follicle stimulating hormone (FSH), luteinizing hormone (LH) and total testosterone (TT) were collected. Successful sperm retrieval (SSR) was obtained in 36 men of total (43.9%).(Table 1) SSR was 29.5% in NOA patients, while OA men, underwent a previous failure TEFNA, had SR in 86% of cases. Mean LH was 6.55 IU/L, TT 4.70 ng/ml, testicular volume (TV) right 13.7 ml and TV left 13.6 ml. FSH mean was 13.45 IU/L in patients with negative SR and 8.18 IU/L in men with SSR. According to histology, 20.7% had Normal Spermatogenesis (NS), 35.3% Hypospermatogenesis (H), 35.3% Maturation Arrest (MA) and 8.5% Sertoli-Cell-Only-Syndrome (SCOS). SSR was 88.2% in patients with NS, 24.1% in the MA group and 48.27% in patients with Hypospermatogenesis, while negative SR was reported in SCOS patients. 2 patients with NS and unsuccessful SR, underwent a second TESE, of whom one had SR and cryopreservation.

Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia.

Non-obstructive azoospermia (NOA) is one the many causes of male infertility (10 %) resulting from testicular failure. Multiple testicular biopsies fail to find mature sperm in at least 50 % of cases Therefore; hunting for sensitive and specific biomarkers of spermatogenesis that could better determine the fertility status in NOA can lead to improved management of male infertility. Therefore, we evaluated sperm production through analyses of germ cell-specific transcripts (DAZ, TSPY1, SPTRX3 and SPTRX1) in semen and testicular biopsies of men with azoospermia. We collected semen (N=83) and testis biopsies (N=31) from men with non-obstructive azoospermia. We later extracted RNA and synthesized cDNA using washed semen precipitate and testicular tissues. We also performed semi-nested PCR with designed specific primers. Using H&E method, an expert pathologist performed the histopathological evaluation. Having categorized the patients into three groups based on histopathological results, we calculated the agreement between molecular results of semen and tissues with histopathological findings for each patient using Kappa statistical test. Molecular findings of precipitated semen and testicular tissues were in disagreement with histopathological results in most cases. Molecular analysis of testis biopsies showed significant difference (Kappa coefficient=0.009, P value=0.894) with histopathological results; TSPY1, DAZ, SPTRX3 and SPTRX1 were respectively detected in 94 %, 94 %, 17.6 % and 52.9 % of men diagnosed with germ cell aplasia. Molecular analysis of semen does not provide sufficient sensitivity and specificity to be used as a screening test at the present time, but it is a useful adjunct to histopathological methods in men with NOA. Spermatid/sperm specific transcripts indicated the possibility to find mature sperm following repeated multiple testicular sperm extraction (TESE) or microdisection TESE (mTESE).

Is ex vivo microdissection testicular sperm extraction indicated for infertile men undergoing radical orchiectomy for testicular cancer? Case report and literature review

To report a case of an infertile man with nonobstructive azoospermia who underwent simultaneous radical orchiectomy for testicular cancer and testicular sperm extraction (TESE) for preservation of fertility. Case report and literature review. University teaching hospital. A couple being treated for infertility. Radical orchiectomy with simultaneous TESE. Sperm retrieval, histologic evaluation of archived testicular pathology slides. We retrieved 20 spermatozoa from the multiple random TESE samples obtained at radical orchiectomy. Histologic evaluation of the archived testicular pathology slides revealed that the testis contained several foci of active spermatogenesis, suggesting that a significantly greater number of spermatozoa would likely have been retrieved had a microdissection TESE been performed instead of the multiple TESEs. We propose that microdissection TESE should be considered the preferred sperm retrieval technique at the time of radical orchiectomy in men with coexistent nonobstructive azoospermia and testicular cancer.

Detection of Y Chromosome Microdeletion is Valuable in the Treatment of Patients With Nonobstructive Azoospermia and Oligoasthenoteratozoospermia: Sperm Retrieval Rate and Birth Rate

PurposeWe evaluated clinical characteristics, sperm retrieval rates, and birth rates in a relatively large number of infertile patients with Y chromosome microdeletions.Materials and MethodsWe retrospectively reviewed clinical data from 213 patients with nonobstructive azoospermia (NOA) and 76 patients with oligoasthenoteratozoospermia (OATS) who were tested for Y chromosome microdeletion from March 2004 to June 2011.ResultsOf the 289 patients, 110 patients presented with Y chromosome microdeletion and 179 patients presented with no microdeletion. Among the patients with Y chromosome microdeletions, 83/110 (75.4%) were NOA patients and 27/110 (24.5%) were OATS patients. After subdividing the patients with Y chromosome microdeletion, 29 had azoospermia factor (AZF)b-c microdeletion and 81 had AZFc microdeletion. The sperm retrieval rate was similar between patients with Y chromosome microdeletion and those with no microdeletion (26.6% vs. 25.6%, p=0.298) after multiple testicular sperm extraction (TESE). Excluding 53 patients who did not undergo TESE, 30 patients were analyzed. All of the 9 men with AZFb-c microdeletion had a complete absence of sperm despite multiple TESE. However, multiple TESE was successful for 9 of 21 patients with only AZFc microdeletion (p=0.041). Twenty patients with Y chromosome microdeletion gave birth.ConclusionsIn NOA and OATS patients, no significant difference in the sperm retrieval rate was shown between patients with Y chromosome microdeletion and those with no microdeletion. Patients with short Y chromosome microdeletion such as AZFc microdeletion have better prognoses for sperm retrieval and an increased chance of conception than do patients with larger microdeletions such as AZFb-c microdeletion.

Success rate of microsurgical multiple testicular sperm extraction and sperm presence in the ejaculate in korean men with y chromosome microdeletions.

PurposeWe assessed the frequency of azoospermia factor a (AZFa), AZFb, and AZFc deletions and examined correlations between the deletion sites and the success rates of sperm presence within the ejaculate and surgical sperm retrieval in Korean men.Materials and MethodsA total of 1,919 azoospermic and severely oligozoospermic men were assessed for Y chromosome microdeletions. Among them, 168 men with AZF deletions were identified and their medical records were reviewed.ResultsOf the total 168 men with AZF deletions, there were 13 with AZFa, 10 with AZFb, 95 with AZFc, 37 with AZFbc, and 13 with AZFabc deletions. Of the 95 men with isolated AZFc deletion, 51 had the presence of sperm in the ejaculate. Of the infertile men with any other deletion, however, only two patients (one man with AZFb deletion and another with AZFbc deletion) showed the presence of sperm in the ejaculate. The success rates for surgical sperm retrieval were 7.1% (1/14) in men with AZFbc deletion and 54.8% (17/31) in the isolated AZFc deletion group. No sperm was obtained from the patients with AZFa or AZFb deletions who underwent microsurgical sperm retrieval. In the isolated AZFc deletion group, there were significant differences between azoospermic and severely oligozoospermic patients in terms of testicular volume and serum levels of follicle-stimulating hormone and luteinizing hormone, whereas no significant differences were found when the group was divided by surgical sperm retrieval outcomes.ConclusionsDeletions of the AZFa and AZFb regions are associated with severe spermatogenetic impairment. However, more than half of men with an AZFc deletion had sperm within the ejaculate or testis for in vitro fertilization with intracytoplasmic sperm injection.

Predictive factors of successful testicular sperm recovery in non-obstructive azoospermia patients.

Recovery of testicular spermatozoa from non-obstructive azoospermic patients for intracytoplasmic sperm injection (ICSI) is a recent advance in the treatment of male infertility. The purpose of this study is to identify predictive factors for sperm recovery in non-obstructive azoospermic patients. A total of 178 men with non-obstructive azoospermia had multiple testicular sperm extraction (TESE) procedures to recover spermatozoa for intracytoplasmic sperm injection (ICSI) from June 1996 to February 1999. Testicular volume, serum follicle stimulating hormone (FSH) level and testicular histology were examined as positive predictive factors for sperm recovery. Testis biopsies were categorized as severe hypospermatogenesis, maturation arrest and Sertoli cell-only syndrome based on the most advanced pattern of spermatogenesis seen on histology. Sperm retrieval success rates for the patients in three histopathological categories were compared. Spermatozoa were successfully recovered in 94 of 178 (52.8%) men. Sperm were retrieved in 13 of 80 (16.3%) with Sertoli cell-only syndrome, 15 of 24 (62.5%) with maturation arrest, and 66 out of 74 (89.2%) with severe hypospermatogenesis. Spermatozoa recovery has no correlation with testicular volume or serum FSH level. When compared against Sertoli cell-only syndrome, the odds of sperm retrieval success rate was 44.3 times higher in severe hypospermatogenesis and 8.4 times in maturation arrest. These results demonstrate meaningful correlation between successful testicular sperm recovery and testis histopathology. Only testicular histopathology can be used as a predictor of successful sperm recovery.

Analysis of Simultaneous Testicular Histopathology, Wet Preparation, Touch Print Cytology and Sperm Recovery in Nonobstructive Azoospermic Males

Objective: Multiple testicular sperm extraction may enhance the diagnostic accuracy of testicular failure and increases the number of sperm cells retrieved. This study questions how many non-obstructive azoospermic (NOA) males generate sufficient number of spermatozoa to be used in ICSI. In order to predict the sperm presence or absence in NOA males, testicular histopathology, wet preparation and touch print cytology were performed simultaneously and the results were compared.

Expression pattern of germ cell–specific genes in the testis of patients with nonobstructive azoospermia: usefulness as a molecular marker to predict the presence of testicular sperm

Objective: To analyze the expression pattern of testis-specific genes of patients with various spermatogenic defects and their usefulness as a molecular marker to predict the presence of testicular spermatozoa in patients with nonobstructive azoospermia undergoing IVF. Design: Prospective, controlled study. Setting: Hospital-based infertility research laboratory. Patient(s): Fifty-eight men with azoospermia or severe oligozoospermia. Intervention(s): Testicular biopsy was done in the patients with obstructive or nonobstructive azoospermia, including Sertoli cell–only syndrome, maturation arrest, severe hypospermatogenesis, and normal spermatogenesis. Main Outcome Measure(s): Reverse transcriptase polymerase chain reaction was performed using 1 μg of total RNA extracted from testicular tissues. Three pairs of primers were used for amplification of male germ cell–specific genes ( DAZ, transcribed in male germ cells; PGK2, in late spermatocytes and spermatids; protamine-2, in spermatids) as molecular markers. Testicular sperm was obtained by multiple testicular sperm extraction. Result(s): The DAZ, PGK2, and protamine-2 genes were expressed in 38, 30, and 21 of the 43 patients with nonobstructive azoospermia, respectively. Testicular spermatozoa were successfully extracted in 4 of 43 patients with nonobstructive azoospermia with the use of multiple testicular sperm extraction. Detection of protamine-2 transcripts predicted the presence or absence of spermatozoa in the testicular tissue in 39 of 43 patients (91%). Conclusion(s): Expression of the protamine-2 gene may be a useful molecular marker to predict the presence of testicular sperm in patients with nonobstructive azoospermia.

Multiple Testicular Sampling in Non-Obstructive Azoospermia-Is it Necessary?

Spermatogenesis may be focal in non-obstructive azoospermia. The present study was conducted to determine whether the performance of multiple, rather than a single testicular sample contributes to obtaining spermatozoa in amounts sufficient for fertilization and cryopreservation in non-obstructive, azoospermic patients. Furthermore, the aim was to clarify the significance of location for retrieval from the testis in such cases. Three biopsies were taken from identical locations in 55 testes of 29 men with non-obstructive azoospermia: (i) the rete testis region, ii) the midline, and (iii) the proximal region of the testis. When sperm cells were detected, they were used for intracytoplasmic sperm injection (ICSI), and the remainder were then cryopreserved in as many aliquots as possible (adjusted for ICSI procedure). Spermatozoa were found in 28 testes (50.9%) of 18 men (62.1%). In the testes from which spermatozoa were obtained, they were present in three, two or one locations in 15 (53.6%), five (17.9%) and eight (28.6%) cases respectively. The possibility of finding spermatozoa was not influenced by the location in the testis. Multiple testicular sperm extraction is recommended in cases of non-obstructive azoospermia, since it may enhance diagnostic accuracy of absolute testicular failure and increase the number of sperm cells retrieved.

Multiple testicular sampling in non-obstructive azoospermia--is it necessary?

Spermatogenesis may be focal in non-obstructive azoospermia. The present study was conducted to determine whether the performance of multiple, rather than a single testicular sample contributes to obtaining spermatozoa in amounts sufficient for fertilization and cryopreservation in non-obstructive, azoospermic patients. Furthermore, the aim was to clarify the significance of location for retrieval from the testis in such cases. Three biopsies were taken from identical locations in 55 testes of 29 men with non-obstructive azoospermia: (i) the rete testis region, ii) the midline, and (iii) the proximal region of the testis. When sperm cells were detected, they were used for intracytoplasmic sperm injection (ICSI), and the remainder were then cryopreserved in as many aliquots as possible (adjusted for ICSI procedure). Spermatozoa were found in 28 testes (50.9%) of 18 men (62.1%). In the testes from which spermatozoa were obtained, they were present in three, two or one locations in 15 (53.6%), five (17.9%) and eight (28.6%) cases respectively. The possibility of finding spermatozoa was not influenced by the location in the testis. Multiple testicular sperm extraction is recommended in cases of non-obstructive azoospermia, since it may enhance diagnostic accuracy of absolute testicular failure and increase the number of sperm cells retrieved.

Testicular sperm extraction for nonobstructive azoospermia: results of a multibiopsy approach with optimized tissue dispersion

Objectives. Testicular sperm extraction (TESE) is an effective procedure to retrieve sperm from some men with nonobstructive azoospermia (NOA). To optimize treatment effectiveness, we have reviewed our experience with TESE for NOA to better understand technical factors needed for sperm retrieval and lead to an optimized approach to TESE. Methods. Eighty-one men with confirmed NOA underwent attempted TESE using an open technique under optical magnification. Each testis sample was dispersed and examined in the operating room. Sequential biopsy attempts were made until sperm were visualized or until further biopsies were thought to jeopardize testicular blood flow. In 20 patients, standard biopsy and initial mechanical dispersion of the seminiferous tubules were compared with the passage of tissue through a 24-gauge angiocatheter after initial dispersion to quantitate spermatozoal yield. Results. Overall, 47 (58%) of 81 patients who underwent TESE had direct intraoperative visualization of spermatozoa. The average number of biopsy attempts for all patients was 8.9 and for patients with sperm isolated 6.4 ( P = 0.002). Passage of the testicular tissue suspension through a 24-gauge angiocatheter increased sperm retrieval in matched tissue specimens from 83,000 to 390,000 or 470% over that achieved with standard dispersion alone ( P = 0.005). An initial, substantive tissue biopsy revealed sperm in only 23 (28%) of 81 patients. Using this approach with sequential biopsies under optical magnification, no patient had evidence of testis injury or devascularization. Conclusions. Because multiple TESE procedures can cause transient and permanent alterations in testicular function, it is imperative to perform TESE as safely and as efficiently as possible. We suggest that open TESE with optical magnification provides a safe method of retrieving sperm. A single biopsy for extraction is inadequate to detect spermatozoa for men with NOA. Use of the needle dispersion technique with passage of testicular tissue through an angiocatheter enhances detection of sperm and could potentially reduce the need for subsequent biopsies. An algorithm to minimize biopsies and allow sperm retrieval is presented.