Microdissection Testicular Sperm Extraction Research Articles

MP30-06 A UNIQUE EPIGENETIC-BASED TEST TO ACCURATELY PREDICT SUCCESSFUL TESE AND mTESE OUTCOMES

You have accessJournal of UrologyCME1 Apr 2023MP30-06 A UNIQUE EPIGENETIC-BASED TEST TO ACCURATELY PREDICT SUCCESSFUL TESE AND mTESE OUTCOMES Tim Jenkins, Ryan Barney, Kelaney Stalker, Ryan Miller, Kristin Brogaard, Jordan Kassab, and Larry Lipshultz Tim JenkinsTim Jenkins More articles by this author , Ryan BarneyRyan Barney More articles by this author , Kelaney StalkerKelaney Stalker More articles by this author , Ryan MillerRyan Miller More articles by this author , Kristin BrogaardKristin Brogaard More articles by this author , Jordan KassabJordan Kassab More articles by this author , and Larry LipshultzLarry Lipshultz More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Affecting 1% of men, non-obstructive azoospermia (NOA) is the absence of sperm in the ejaculate, and is the most severe form of male infertility. Currently, men with NOA have decreased chances of conceiving a child of their own. Despite recent progress in reproductive medicine, men with NOA have essentially one invasive and expensive procedure to isolate sperm (microdissection testicular sperm extraction or mTESE). Unfortunately, mTESE has a ∼50% failure rate. Here we present the use of an innovative DNA methylation-based technology to non-invasively predict the presence of sperm in the testicle of men with NOA. METHODS: At a single academic center biopsies (N=18) and azoospermic semen samples (N=5) were procured from consented men who had undergone a testicular aspirate, biopsy, or mTESE procedure. Utilizing new epigenetic technology, the biopsies and semen samples were analyzed for sperm-specific DNA. Specifically, these samples were analyzed for a sequence of DNA that can distinguish sperm cells from all other somatic cells via a unique methylation pattern. The quantity of sperm-specific DNA can be used to determine the absence (men with <0.25 ratio of sperm to somatic DNA) or presence (>0.5 ratio of sperm to somatic DNA) of sperm in the testicles. These ratios were defined for each sample and compared to clinical outcomes of the TESA, TESE or mTESE procedures. RESULTS: Out of the 18 biopsies, 13 had >0.5 ratios of sperm DNA to somatic cell DNA in the testicular biopsies. All of these men were found to have sperm in their biopsies as defined from the clinical notes. 3 of the 18 men had <0.25 ratios of sperm DNA to somatic cell DNA. Again, consistent with previous analysis, none of these men had sperm in their biopsies. The remaining 2 men had ratios between 0.26 – 0.49, with one having sperm in his biopsy and one without sperm. Of the 5 azoospermic semen samples with associated TESA results, 3 has ratios of <0.25 and all 3 had no sperm in the corresponding TESA samples. 2 semen samples had ratios of >0.5 and were confirmed to have sperm in the TESA sample. CONCLUSIONS: In the analysis of testicular biopsies and semen samples, the sperm-specific DNA methylation ratios of <0.25% or >0.5% were 100% predictive of either the absence or presence of sperm in the testicle. With these data the next steps of our work will focus on continued analysis of sperm-specific cfDNA in seminal plasma to predict the presence or absence of sperm prior to a testicular extraction procedure. The ultimate goal will be to develop a non-invasive and cost-effective way to avoid unnecessary surgical procedures. Source of Funding: Venture Capital © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e393 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Tim Jenkins More articles by this author Ryan Barney More articles by this author Kelaney Stalker More articles by this author Ryan Miller More articles by this author Kristin Brogaard More articles by this author Jordan Kassab More articles by this author Larry Lipshultz More articles by this author Expand All Advertisement PDF downloadLoading ...

MP43-04 EFFECTS OF AGE ON TESTICULAR FUNCTION AND THE SPERM RETRIEVAL RATE IN MEN WITH NONOBSTRUCTIVE AZOOSPERMIA

You have accessJournal of UrologyCME1 Apr 2023MP43-04 EFFECTS OF AGE ON TESTICULAR FUNCTION AND THE SPERM RETRIEVAL RATE IN MEN WITH NONOBSTRUCTIVE AZOOSPERMIA Shoichiro Iwatsuki, Yukihiro Umemoto, Takumi Kito, Tomoki Takeda, Satoshi Nozaki, and Takahiro Yasui Shoichiro IwatsukiShoichiro Iwatsuki More articles by this author , Yukihiro UmemotoYukihiro Umemoto More articles by this author , Takumi KitoTakumi Kito More articles by this author , Tomoki TakedaTomoki Takeda More articles by this author , Satoshi NozakiSatoshi Nozaki More articles by this author , and Takahiro YasuiTakahiro Yasui More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003289.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: We previously reported an increase in the age of men with nonobstructive azoospermia (NOA) (AUA Annual Meeting 2020). It is important to determine age-induced effects on testicular function in men with NOA. We investigated age-induced changes in testicular endocrine function and the sperm retrieval rate (SRR) during microdissection testicular sperm extraction in men with NOA. METHODS: We investigated 323 men with idiopathic NOA. Patients were categorized into the following age-based groups: 20 s (20–30 years, n=64), 30 s (30–40 years, n=201), and >40 s (>40 years, n=58). We retrospectively recorded testicular volume, serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), total testosterone (TT), and anti-Mullerian hormone (AMH) levels, and SRRs. RESULTS: Testicular volumes in groups 20 s, 30 s, and >40 s were 9.5±4.3 vs. 9.1±4.0 vs. 7.9±3.9 mL, p<0.05 (Jonckheere-Terpstra trend test). Although we observed no intergroup differences in serum LH and TT levels, FSH levels significantly increased with age (22.4±13.3 vs. 26.9±13.2 vs. 26.8±15.3 mIU/mL, p<0.05) as did serum AMH levels (4.4±4.3 vs. 7.9±6.3 vs. 9.2±7.0 ng/mL, p<0.01). SRRs in the 20 s, 30 s, and 40 s groups were 12.5 vs. 28.4 vs. 36.2%, p<0.05 (Cochran-Armitage trend test). CONCLUSIONS: We observed age-induced testicular atrophy and increased serum FSH and AMH levels, which highlight age-induced testicular function decline in men with NOA and suggest that NOA may be progressive despite favorable SRRs in older men. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e602 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Shoichiro Iwatsuki More articles by this author Yukihiro Umemoto More articles by this author Takumi Kito More articles by this author Tomoki Takeda More articles by this author Satoshi Nozaki More articles by this author Takahiro Yasui More articles by this author Expand All Advertisement PDF downloadLoading ...

MP30-05 CONCORDANCE OF TESTIS HISTOLOGY AMONG MEN UNDERGOING BILATERAL TESTICULAR SPERM EXTRACTION PROCEDURES

You have accessJournal of UrologyCME1 Apr 2023MP30-05 CONCORDANCE OF TESTIS HISTOLOGY AMONG MEN UNDERGOING BILATERAL TESTICULAR SPERM EXTRACTION PROCEDURES Jessica Marinaro, Jonathan Gal, and James Kashanian Jessica MarinaroJessica Marinaro More articles by this author , Jonathan GalJonathan Gal More articles by this author , and James KashanianJames Kashanian More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: For men undergoing a microdissection testicular sperm extraction (mTESE) procedure, if sperm are not found in one testis, a bilateral procedure is typically performed; however, this invasive dissection may have adverse effects. While testicular histology may predict sperm retrieval rates (SRRs), little is known about the concordance of histology between testes. Since this may inform the utility of performing a bilateral versus unilateral dissection, our objective was to assess the rate of histological concordance between left and right testis biopsies and as well as the overall sperm retrieval rate (SRR) for these patients. METHODS: We retrospectively reviewed electronic medical records using CPT codes 55899 and 54505 for 1 surgeon from 6/2016 to 8/2022. Men were included if they underwent a testicular sperm extraction (TESE) or mTESE procedure. Biopsies were evaluated by 4 pathologists at our institution. Histology was classified as Sertoli-cell only (SCO), maturation arrest (MA; early or late), active spermatogenesis (AS), or tubular atrophy (TA). Each pattern was reported as a percentage contributing to a total of 100%. A successful sperm retrieval was defined as the presence of sperm after processing by an andrology lab. RESULTS: Of 128 men who underwent a TESE or mTESE, 38 (29.7%) had a bilateral procedure. Mean demographics for these 38 men included an age at surgery of 39.4 ± 7.0 years, baseline testosterone 400.1 ± 198.0 ng/dL, baseline FSH 18.7 ± 14.9 mIU/mL, right testis volume 10.1 ± 6.4 mL, and left testis volume 10.2 ± 6.4 mL. Thirty-one men (31/38, 81.6%) had biopsies sent for histology from both testes. Of these, 9 (29.0%) had the same histology in the same proportions bilaterally, 14 (45.2%) had the same predominant histology but in variable proportions, and 8 (25.8%) had discordant predominant histology. Ten of 31 men (32.3%) had sperm retrieved; of these, 6 (60%) had no evidence of spermatogenesis on either histology specimen. CONCLUSIONS: While it is known that there is often heterogeneity within the testis, this study confirms that there is also significant heterogeneity between each testis for >25% of men. Though histology may help to predict SRRs, this study emphasizes that histology alone is not reliable in determining which men will have sperm present after processing by an andrology lab. Ultimately, while histology may provide patients and providers with useful information on testicular architecture, it should not be the only factor in clinical decision making, such as who should undergo a bilateral procedure or repeat procedure. Source of Funding: Authors JM and JG are supported in part by the Frederick J. and Theresa Dow Wallace Fund of the New York Community Trust. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e392 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jessica Marinaro More articles by this author Jonathan Gal More articles by this author James Kashanian More articles by this author Expand All Advertisement PDF downloadLoading ...

MP01-09 GENE DOSAGE CHANGES IN CRYPTORCHID MEN WITH NONOBSTRUCTIVE AZOOSPERMIA

You have accessJournal of UrologyCME1 Apr 2023MP01-09 GENE DOSAGE CHANGES IN CRYPTORCHID MEN WITH NONOBSTRUCTIVE AZOOSPERMIA Aaron Brant, Caroline Kang, Anna Mielnik, Marisol O'Neill, Meade Haller, Peter Schlegel, and Dolores Lamb Aaron BrantAaron Brant More articles by this author , Caroline KangCaroline Kang More articles by this author , Anna MielnikAnna Mielnik More articles by this author , Marisol O'NeillMarisol O'Neill More articles by this author , Meade HallerMeade Haller More articles by this author , Peter SchlegelPeter Schlegel More articles by this author , and Dolores LambDolores Lamb More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003212.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Copy number variations (CNVs) are structural chromosomal defects causing microdeletions or microduplications. CNVs in the dosage-sensitive genes ADCY2, ADCY9, CRKL, KCTD13, RBFOX2, and ZEB2 occur more frequently in men with genitourinary anomalies such as cryptorchidism and hypospadias compared to the general male population. To evaluate the association of these CNVs with spermatogenic failure, the prevalence of CNVs in men with history of cryptorchidism who underwent microdissection testicular sperm extraction (mTESE) for non-obstructive azoospermia (NOA) was defined. METHODS: Whole blood DNA samples were collected from 101 men with cryptorchidism and NOA who presented for (mTESE) and two fertile control men. TaqMan quantitative polymerase chain reaction was used to assess copy numbers of ADCY2, ADCY9, CRKL, KCTD13, RBFOX2, and ZEB2 with each reaction performed in triplicate. Relative quantification values were obtained using CopyCaller Software. Fisher’s exact test was used to compare CNVs in cryptorchid men with NOA compared to CNV prevalence in the general male population from the Database of Genomic Variants. RESULTS: Of the 101 patients, 44 (44%) had bilateral cryptorchidism, 57 (56%) had unilateral cryptorchidism, and 51 (50%) had sperm retrieved with mTESE. Six patients (5.9%) had CNVs detected. Two men had microdeletions of CRKL, one had a microdeletion of ZEB2, and one had a microduplication of ADCY2. Two men with bilateral cryptorchidism had multiple CNVs detected: one had microdeletions of ADCY2, ADCY9, and RBFOX2, and one had microdeletions of ADCY9, CRKL, ZEB2 and a microduplication of KCTD13. Four of the six patients with detected CNVs (67%) had sperm retrieved on mTESE. Compared the general male population, men with cryptorchidism and NOA had higher prevalence of CNVs in ADCY2 (p=0.002), ADCY9 (p=0.004), CRKL (p<0.001), and ZEB2 (p<0.001). CONCLUSIONS: Approximately 6% of adult men with a history of cryptorchidism and NOA had gene dosage changes in ADCY2, ADCY9, CRKL, KCTD13, RBFOX2, or ZEB2, with two-thirds of these men having successful sperm retrieval on mTESE. As many of these CNVs are syndromic, genetic counseling may be warranted for men found to carry them. The functional consequences of these gene dosage changes on adult spermatogenesis remain to be fully elucidated. Source of Funding: n/a © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e5 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Aaron Brant More articles by this author Caroline Kang More articles by this author Anna Mielnik More articles by this author Marisol O'Neill More articles by this author Meade Haller More articles by this author Peter Schlegel More articles by this author Dolores Lamb More articles by this author Expand All Advertisement PDF downloadLoading ...

PD18-07 SERUM ANTI-MÜLLERIAN HORMONE IN RELATION TO SPERM RETRIEVAL RATE AND TESTICULAR HISTOPATHOLOGY IN NON-OBSTRUCTIVE AZOOSPERMIA

You have accessJournal of UrologyCME1 Apr 2023PD18-07 SERUM ANTI-MÜLLERIAN HORMONE IN RELATION TO SPERM RETRIEVAL RATE AND TESTICULAR HISTOPATHOLOGY IN NON-OBSTRUCTIVE AZOOSPERMIA Tomoki Takeda, Shoichiro Iwatsuki, Satoshi Nozaki, Hidenori Nishio, Hiroki Kubota, Yasue Kubota, Hiroyuki Kamiya, Hidetoshi Akita, Kentaro Mizuno, Yutaro Hayashi, Yukihiro Umemoto, and Takahiro Yasui Tomoki TakedaTomoki Takeda More articles by this author , Shoichiro IwatsukiShoichiro Iwatsuki More articles by this author , Satoshi NozakiSatoshi Nozaki More articles by this author , Hidenori NishioHidenori Nishio More articles by this author , Hiroki KubotaHiroki Kubota More articles by this author , Yasue KubotaYasue Kubota More articles by this author , Hiroyuki KamiyaHiroyuki Kamiya More articles by this author , Hidetoshi AkitaHidetoshi Akita More articles by this author , Kentaro MizunoKentaro Mizuno More articles by this author , Yutaro HayashiYutaro Hayashi More articles by this author , Yukihiro UmemotoYukihiro Umemoto More articles by this author , and Takahiro YasuiTakahiro Yasui More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003273.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Anti-Müllerian hormone (AMH) is secreted by Sertoli cells. Although AMH has been suggested as a marker of spermatogenesis, the relationship between serum AMH and spermatogenesis in non-obstructive azoospermia (NOA) is unclear. Therefore, we investigated the relationship between serum AMH and sperm retrieval rate (SRR), testicular histopathology in NOA. METHODS: The study population consisted of 237 patients with NOA who underwent microdissection testicular sperm extraction and had their serum AMH levels measured between January 2015 and December 2021. NOA patients were classified into serum AMH levels <0.1 (detection limit, n=27), 0.1-4 (n=76), 4-8 (n=58) and ≥8 ng/ml (n=76), and SRRs were compared. In addition, NOA patients were classified into TH (tubular hyalinization, n=15), SCO (Sertoli cell only syndrome, n=132), MA (maturation arrest, n=57) and HS (hypospermatogenesis, n=33) groups. The percentage of patients with serum AMH levels <0.1 ng/ml and serum AMH levels were compared. RESULTS: SRR were 3.7%, 40.8%, 36.2% and 23.7% for serum AMH levels <0.1, 0.1-4, 4-8 and ≥8 ng/ml, respectively (Figure 1A), with a significant difference between serum AMH levels <0.1 ng/ml and the other groups (p<0.01). The percentages of patients with serum AMH levels below 0.1 ng/ml were 86.7%, 10.6%, 0% and 0% in TH, SCO, MA and HS groups, respectively (Figure 1B), with a significant difference between TH and the other groups (p<0.01). Serum AMH levels were 0.11±0.04, 6.91±6.59, 7.75±7.09 and 5.77±3.58 ng/ml for the four groups, respectively (Figure 1C), with a significant difference between TH and the other groups (p<0.01). CONCLUSIONS: When the serum AMH value ≥0.1 ng/ml, the higher the serum AMH value, the lower the SRR. On the other hand, serum AMH levels <0.1 ng/ml may suggest hyalinization of seminiferous tubules, loss of Sertoli cells and low possibility of sperm retrieval. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e505 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Tomoki Takeda More articles by this author Shoichiro Iwatsuki More articles by this author Satoshi Nozaki More articles by this author Hidenori Nishio More articles by this author Hiroki Kubota More articles by this author Yasue Kubota More articles by this author Hiroyuki Kamiya More articles by this author Hidetoshi Akita More articles by this author Kentaro Mizuno More articles by this author Yutaro Hayashi More articles by this author Yukihiro Umemoto More articles by this author Takahiro Yasui More articles by this author Expand All Advertisement PDF downloadLoading ...

MP30-07 EVALUATION OF MTESE OUTCOMES IN PATIENTS WITH PARTIAL AZFb Y CHROMOSOME MICRODELETIONS

You have accessJournal of UrologyCME1 Apr 2023MP30-07 EVALUATION OF MTESE OUTCOMES IN PATIENTS WITH PARTIAL AZFb Y CHROMOSOME MICRODELETIONS Jonathan Gal, Jessica Marinaro, Anna Mielnik, and Peter Schlegel Jonathan GalJonathan Gal More articles by this author , Jessica MarinaroJessica Marinaro More articles by this author , Anna MielnikAnna Mielnik More articles by this author , and Peter SchlegelPeter Schlegel More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Y chromosome microdeletions (YCMD) are a common cause of non-obstructive azoospermia. While men with deletions in the azoospermia factor C (AZFc) region have a high likelihood of having sperm in the ejaculate, or sperm retrievable with effective microdissection testicular sperm extraction (mTESE) procedure, those with complete deletions in the AZFa or AZFb regions have a poor prognosis for identification of sperm. Men with AZFa deletions typically do not have any germ cells, and no men, to-date, with complete AZFb deletions have had usable sperm identified. Partial deletions of AZFb appear to portend a different prognosis for fertility potential. Our objective was to evaluate semen parameters, sperm retrieval rates and fertility outcomes in men with partial deletions of the AZFb region. METHODS: We retrospectively reviewed 468 men with YCMDs who underwent testing at our institution from 4/1996 to 8/2022. Of these, we identified 20 men with partial AZFb deletions, defined as a deletion of some but not all tested segments from sY1227 to sY1291. Of these, 8 had documented semen analysis and underwent mTESE by a single, high-volume surgeon (PS). Sperm retrieval rate (SRR) was defined as the presence of mature sperm identified after processing by a trained andrologist. RESULTS: Of the 8 men with a partial AZFb deletion, 7 (87.5%) had a sY117-sY1291 deletion, and one (12.5%) had an sY1227-sY1192 deletion. All 8 men (100%) were azoospermic. Mean testosterone levels were 450.5 ± 350.5 (ng/dL), Mean FSH 6.5±2.8 (mIU/mL). After mTESE, 1 of 7 men (14%) with an sY117-sY1291 deletion was found to have sperm, and the 1 man with a sY1227-sY1192 deletion was found to have round spermatids on histology. Overall, the SRR was 12%. All pathology specimens (n=8, 100%) demonstrated maturation arrest. CONCLUSIONS: In this large series, we demonstrate that while patients with partial AZFb deletions are azoospermic, it is possible to find sperm at least for some men with mTESE. Distinguishing partial from complete AZFb deletions may have prognostic value. Further work is needed to determine which men with partial AZFb deletions are more likely to have a successful mTESE procedure. Source of Funding: Author JG is supported in part by the Frederick J. and Theresa Dow Wallace Fund of the New York Community Trust © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e393 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jonathan Gal More articles by this author Jessica Marinaro More articles by this author Anna Mielnik More articles by this author Peter Schlegel More articles by this author Expand All Advertisement PDF downloadLoading ...

Factors influencing the sperm retrieval rate of microdissection testicular sperm extraction in patients with nonmosaic Klinefelter syndrome.

To investigate the factors affecting the sperm retrieval rate of microdissection testicular sperm extraction (micro-TESE) in patients with nonmosaic Klinefelter syndrome (KS), 64 patients with nonmosaic KS who underwent micro-TESE in the Center for Reproductive Medicine of Peking University Third Hospital (Beijing, China) between January 2016 and December 2017 were included in the study. Data on medical history, physical examination and laboratory examination results, and micro-TESE outcomes were collected. Patients were divided into two groups according to micro-TESE outcomes. The following factors were compared between the two groups by the Mann ‒ Whitney U test or Student's t-test based on the distribution (nonnormal or normal) of the factors: age, testicular size, follicle-stimulating hormone level, luteinizing hormone level, testosterone level, and anti-Müllerian hormone level. The overall success rate of sperm retrieval was 50.0%. Correlation analysis showed that testicular volume was positively correlated with testosterone level. Using a logistic regression model, age and anti-Müllerian hormone levels were found to be better predictors for the sperm retrieval rate than the other parameters.

Touch print smear of testicular tissue with thionine stain for intraoperative diagnosis in azoospermic men.

To compare the diagnostic value of testicular tissue touch print smear (TPS) conducted on azoospermic patients with results obtained from histopathology and in vitro fertility (IVF) lab findings. Microdissection testicular sperm extraction was performed on a group of 148 azoospermic patients and testicular samples obtained intraoperatively. Using TPS, the samples were smeared onto a sterile slide, followed with staining using thionine. The testis tissue bulk samples were also transferred to the IVF lab, and determinations of sperm presence or absence obtained from IVF lab tests were compared with the TPS sample results. Needle testis biopsy was separately performed on a group of 360 azoospermic patients, and results of pathohistology review on the biopsies were further compared with determinations of spermatogenesis stage obtained from TPS for those patients. When compared with IVF lab results, TPS was found to have 100% (126/126) positive predictive value and 95.5% (25/26) negative predictive value for predicting sperm presence or absence, respectively. Furthermore, TPS was further found to have a 93.6% correlation (337 of 360 biopsies) with results of histological diagnoses performed by needle biopsy. Results from histology and TPS for the detection of sperm presence were concordant in 96.1% (346/360) of biopsies. Diagnosis of SCO by TPS shows the highest correlation with histopathology (98.6%), followed by complete spermatogenesis (97.5%), early maturation arrest (78.9%), and late maturation arrest (27.3%). The results support the continued use of TPS in testicular tissue analysis for more rapid assessment of spermatogenesis and for detection of spermatozoa in azoospermic subjects.

Outcomes of Microdissection Testicular Sperm Extraction/Intracytoplasmic Sperm Injection in Cases of Nonobstructive Azoospermia: A Retrospective Study

The outcomes and safety of intracytoplasmic sperm injection (ICSI) using testicular sperm have been controversial. We evaluated ICSI results, pregnancy outcomes, and neonatal health conditions using testicular sperm from patients with obstructive (OA) or nonobstructive (NOA) azoospermia. We further compared the ICSI outcomes after use of fresh versus cryopreserved testicular sperm from men with NOA. We included 314 men with NOA who underwent microdissection testicular sperm extraction (mTESE) and 303 with OA who underwent testicular sperm aspiration; both groups underwent ICSI. Therefore, 101 and 329 ICSI cycles were performed for mTESE and aspirated sperms, respectively. Furthermore, fresh and thawed embryos from both groups were transplanted to evaluate fertilization and pregnancy rates (NOA, 15 fresh and 123 thawed; OA, 59 fresh and 393 thawed). Finally, of the 101 ICSI cycles performed for NOA patients, 56 fresh-sperm cycles and 45 thawed-sperm cycles were performed to evaluate ICSI outcomes. Fertilization rates and two-pronuclear (2PN) fertilization rates were significantly lower in the NOA group than in the OA group. However, the 2PN cleavage rate, the high-quality embryo rate, the blastocyst formation rate, and the available blastocyst rate showed no significant intergroup differences. In addition, the pregnancy outcomes and neonatal health conditions were statistically similar. Finally, compared with thawed sperm, the fresh-sperm group had a higher 2PN fertilization rate and a higher high-quality embryo rate. However, blastocyst formation and available blastocyst rates were similar between the two groups. Patients with NOA achieved the same favorable results in embryo development, clinical pregnancy, and live birth capability as did OA patients. Neonatal conditions were not affected by type of azoospermia (NOA versus OA). For patients with NOA, fresh testicular sperm is superior to frozen-thawed testicular sperm in embryo development as evaluated at the cleavage stage, but we find no superiority evaluating at the stage of blastocyst formation.

Efficacy of stepwise mini-incision microdissection testicular sperm extraction for nonobstructive azoospermia with varied etiologies.

Stepwise mini-incision microdissection testicular sperm extraction (mTESE) is a procedure that attempts to minimize testicular damage. However, the mini-incision approach may vary in patients with different etiologies. Here, we performed a retrospective analysis of 665 men with nonobstructive azoospermia (NOA) who underwent stepwise mini-incision mTESE (Group 1) and 365 men who underwent standard mTESE (Group 2). The results showed that the operation time (mean ± standard deviation) for patients with successful sperm retrieval in Group 1 (64.0 ± 26.6 min) was significantly shorter than that in Group 2 (80.2 ± 31.3 min), with P <0.001. The total sperm retrieval rate (SRR) was 23.1% in our study, and there was no significant difference between Group 1 and Group 2 ( P >0.05), even when the etiologies of NOA were taken into consideration. The results of consecutive multivariate logistic regression analysis (odds ratio [OR]: 0.57; 95% confidence interval [CI]: 0.38-0.87; P =0.009) and receiver operating characteristic (ROC) analysis (area under the ROC curve [AUC]=0.628) showed that preoperative anti-Müllerian hormone (AMH) level in idiopathic NOA patients was a potential predictor for surgical outcomes after initial three small incisions made in the equatorial region without sperm examined under an operating microscope (Steps 2-4). In conclusion, stepwise mini-incision mTESE is a useful technique for NOA patients, with comparable SRR, less surgical invasiveness, and shorter operation time compared with the standard approach. Low AMH levels may predict successful sperm retrieval in idiopathic patients even after a failed initial mini-incision procedure.

Embryo development and live birth resulted from artificial oocyte activation after microdissection testicular sperm extraction with ICSI in patients with non-obstructive azoospermia.

The application of microdissection testicular sperm extraction (micro-TESE) to retrieve the sperm of patients with non-obstructive azoospermia (NOA) has greatly increased. Patients with NOA often have poor quality sperm. Unfortunately, there are few studies on artificial oocyte activation (AOA) performed on patients who successfully retrieved motile and immotile sperm by micro-TESE after intracytoplasmic sperm injection (ICSI). Therefore, this study sought to obtain more comprehensive evidence-based data and embryo development outcomes to aid consultation of patients with NOA who opted to receive assisted reproductive techniques and to determine whether AOA needs to be performed in different motile sperm after ICSI. This retrospective study involved 235 patients with NOA who underwent micro-TESE to retrieve adequate sperm for ICSI between January 2018 and December 2020.A total of 331 ICSI cycles were performed in the 235 couples. Embryological, clinical, and neonatal outcomes were demonstrated comprehensively between motile sperm and immotile sperm using AOA and non-AOA treatment. Motile sperm injection with AOA (group 1) showed significantly higher fertility rate (72.77% vs. 67.59%, p=0.005), 2 pronucleus (2PN) fertility rate (64.33% vs. 60.22%, p=0.036), and miscarriage rate (17.65% vs. 2.44%, p=0.018) compared with motile sperm injection with non-AOA (group 2). Group 1 had comparable available embryo rate (41.29% vs. 40.74%, p=0.817), good embryo rate (13.44% vs. 15.44%, p=0.265), and without an embryo for transfer rate (10.85% vs. 9.90%, p=0.815) compared with group 2. Immotile sperm injection with AOA (group 3) displayed significantly higher fertility rate (78.56% vs. 67.59%, p=0.000), 2PN fertility rate (67.36% vs. 60.22%, p=0.001), without an embryo for transfer rate (23.76% vs. 9.90%, p=0.008), and miscarriage rate (20.00% vs. 2.44%, p=0.014), but significantly lower available embryo rate (26.63% vs.40.74%, p=0.000) and good embryo rate (15.44% vs. 6.99%, p=0.000) compared with group 2. In groups 1, 2, and 3, the rates of implantation (34.87%, 31.85% and 28.00%, respectively; p=0.408), clinical pregnancy (43.87%, 41.00%, and 34.48%, respectively; p=0.360) and live birth (36.13%, 40.00%, and 27.59%, respectively; p=0.194) were similar. For those patients with NOA from whom adequate sperm were retrieved for ICSI, AOA could improve fertilization rate, but not embryo quality and live birth outcomes. For patients with NOA and only immotile sperm, AOA can help achieve acceptable fertilization rate and live birth outcomes. AOA is recommended for patients with NOA only when immotile sperm are injected.

Development of a predictive model for increasing sperm retrieval success by microdissection testicular sperm extraction in patients with nonobstructive azoospermia.

Microdissection testicular sperm extraction (micro-TESE) is widely used to treat nonobstructive azoospermia. However, a good prediction model is required to anticipate a successful sperm retrieval rate before performing micro-TESE. This retrospective study analyzed the clinical records of 200 nonobstructive azoospermia patients between January 2021 and December 2021. The backward method was used to perform binary logistic regression analysis and identify factors that predicted a successful micro-TESE sperm retrieval. The prediction model was constructed using acquired regression coefficients, and its predictive performance was assessed using the receiver operating characteristic curve. In all, 67 patients (sperm retrieval rate: 33.5%) underwent successful micro-TESE. Follicle-stimulating hormone, anti-Müllerian hormone, and inhibin B levels varied significantly between patients who underwent successful and unsuccessful micro-TESE. Binary logistic regression analysis yielded the following six predictors: anti-Müllerian hormone (odds ratio [OR] = 0.902, 95% confidence interval [CI]: 0.821-0.990), inhibin B (OR = 1.012, 95% CI: 1.001-1.024), Klinefelter's syndrome (OR = 0.022, 95% CI: 0.002-0.243), Y chromosome microdeletion (OR = 0.050, 95% CI: 0.005-0.504), cryptorchidism with orchiopexy (OR = 0.085, 95% CI: 0.008-0.929), and idiopathic nonobstructive azoospermia (OR = 0.031, 95% CI: 0.003-0.277). The prediction model had an area under the curve of 0.720 (95% CI: 0.645-0.794), sensitivity of 65.7%, specificity of 72.2%, Youden index of 0.379, and cut-off value of 0.305 overall, indicating good predictive value and accuracy. This model can assist clinicians and nonobstructive azoospermia patients in decision-making and avoiding negative micro-TESE results.

Nomogram based on a circular RNA biomarker for predicting the likelihood of successful sperm retrieval via microdissection testicular sperm extraction in patients with idiopathic non-obstructive azoospermia.

Many circular RNAs (circRNAs) are specifically expressed in the testes and seminal plasma of patients with non-obstructive azoospermia (NOA), highlighting them as potential predictors of microdissection testicular sperm extraction (micro-TESE) outcomes. Although research has indicated that circular RNA monoglyceride lipase (circ_MGLL) is highly expressed in the testicular tissues of patients with NOA, the association between circ_MGLL expression and sperm retrieval outcomes (SROs) in patients with idiopathic non-obstructive azoospermia (iNOA) receiving micro-TESE remains unclear. This single-center, retrospective cohort study enrolled 114 patients with iNOA who underwent micro-TESE at Northwest Women's and Children's Hospital from January 2017 to November 2021.A logistic regression model was used to examine associations between SRO and circ_MGLL expression in testicular tissues, the results of which were used in conjunction with previous findings to establish a nomogram. The predictive performance of the circ_MGLL-based nomogram was evaluated via calibration curves, receiver operating characteristic curves, and decision curve analysis (DCA) using an internal validation method. The generalized additive model indicated that the probability of successful SRO for micro-TESE decreased as circ_MGLL expression increased in testicular tissues. Across the entire cohort, univariate logistic regression analysis revealed that circ_MGLL expression was inversely associated with SRO in patients with NOA. This trend did not change after stratification according to age, body mass index, testicular volume, follicle-stimulating hormone (FSH) level, luteinizing hormone (LH) level, testosterone (T) level, or pathological type (or after adjusting for these confounders) (odds ratio <1, P < 0.001). A nomogram was then generated by integrating circ_MGLL, pathological types, and FSH, LH, and T levels. The circ_MGLL-based predictive model achieved satisfactory discrimination, with an area under the curve of 0.857, and the calibration curves demonstrated impressive agreement. The DCA indicated that the net clinical benefit of the circ_MGLL-based predictive model was greater than that of circ_MGLL alone. circ_MGLL is significantly associated with the SRO of micro-TESE in patients with iNOA. The circ_MGLL-based nomogram developed in the current study can predict successful SRO with high accuracy.

Identification of active spermatogenesis using a multiphoton microscope.

The sperm retrieval rate of microdissection testicular sperm extraction varies from 25% to 60%. Therefore, it is necessary to establish objective selection criteria for identifying seminiferous tubules with spermatozoa. Our aim was to develop a method for identifying spermatogenesis without sectioning testicular tissues. Testicular tissues of 10-week-old normal rats were fixed with 4% paraformaldehyde. Fluorescent labeling of seminiferous tubule nuclei and F-actin was performed, and the specimens were observed without sectioning using a multiphoton microscope. Cryptorchid rats were used as a model lacking elongated spermatids. Multiphoton images were compared with images of normal seminiferous tubules. In addition, seminiferous tubules of 10-week-old normal rats were labeled by testicular interstitial injection of fluorescent probes and observed by a multiphoton microscope without fixation. Terminal deoxynucleotidyl transferase dUTP nick end labeling-stained images of normal and probe-injected testes were compared. In fixed seminiferous tubules, elongated spermatids were identified. In addition, F-actin of apical ectoplasmic specialization was observed around elongated spermatids. Furthermore, spermatogenic stages were identified by an array of nuclei or F-actin. In cryptorchid testes, elongated spermatids and F-actin of the apical ectoplasmic specialization were not observed. In testes injected with fluorescent probes, F-actin of the apical ectoplasmic specialization was observed, and spermatogenic stages were identified without fixation. There was no significant difference in the number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells per seminiferous tubule between normal and probe-injected testes. Seminiferous epithelium could be observed without sectioning of tissues by fluorescent probes and a multiphoton microscope. Active spermatogenesis was observed by labeling F-actin with and without fixation. Moreover, the toxicity of fluorescent probes was limited. Our method has a potential for live imaging of testicular tissue.

Interaction between Host and Microbes in the Semen of Patients with Idiopathic Nonobstructive Azoospermia.

Men with nonobstructive azoospermia (NOA) face the dual problems of low sperm count and low sperm quality. Most men with NOA without a clear cause are classified as having idiopathic NOA (iNOA). Previous studies found that microbes exist in semen, and the semen microbes of NOA men are different from those of normal men. However, the relevant mechanism is not clear. In this study, we answered the three questions of "who is there," "what is it doing," and "who is doing it" by combining 16s rRNA, nontargeted metabolome detection and metabolite traceability analysis. We found that the composition and interaction of seminal plasma microbes in the iNOA group changed. Metabolite traceability analysis and metabolic pathway analysis revealed that microbial abnormalities in the NOA group were closely related to the decrease of microbial degradation of toluene and the increase of metabolism of fructose or mannose. In addition, the metabolic relationship between microbes and the host in male semen in iNOA revealed that such microbes can produce harmful metabolites that affect sperm quality, the microbes compete with sperm for essential nutrients, and their presence reduces sperm production of essential nutrients. IMPORTANCE Idiopathic nonobstructive azoospermia is one of the great challenges in assisted reproductive therapy. Although microdissection testicular sperm extraction technology is currently available, many men with iNOA still face the problem of poor sperm retrieval and poor sperm quality. The role of seminal plasma microbes in male disease has been continuously investigated since semen was demonstrated to harbor commensal microbes. To our knowledge, this is the first detailed description of the microbe-host relationship in iNOA semen. This study is an important complement to research on the treatment and etiology of iNOA and the rationale for our ongoing research.

A dual-center study of predictive factors for sperm retrieval through microdissection testicular sperm extraction and intracytoplasmic sperm injection outcomes in men with non-mosaic Klinefelter syndrome.

This study evaluated the predictors of sperm retrieval (SR) in non-mosaic Klinefelter syndrome (KS) patients undergoing microsurgical testicular sperm extraction (mTESE). The cutoff values of the predictors of SR and overall pregnancy rates after intracytoplasmic sperm injection (ICSI) were analyzed for the positive SR (PSR) cases. The study was a dual-center retrospective study. Overall 118 patients with KS underwent mTESE between January 2011 and July 2021. Clinicopathological factors including comorbidities, endocrine profiles, and testicular volumes were analyzed. A total of 58 patients showed PSR (49.2%) and 60 patients (50.8%) had negative SR (NSR). The mean overall age of the patients was 32.5 years. The NSR patients had a significantly greater prevalence of obesity, diabetes mellitus, and cerebrovascular disease. The PSR group had a significantly higher left testis mean volume (p=0.039). The differences between the two study groups regarding follicular-stimulating hormone, luteinizing hormone, and testosterone variations at 1 and 3 months after mTESE were insignificant. Preoperative mean neutrophil-to-lymphocyte ratio was significantly greater in the NSR group (p=0.011), but the platelet-to-lymphocyte ratio showed no significant difference between the two study groups. A live child birth was achieved in 53.4% of the PSR patients. Multivariate logistic analysis showed that total testicular volume >3.93 mL, left testis volume >1.79 mL, and neutrophil-to-lymphocyte ratio ≤1.82 were significantly associated with PSR. mTESE-ICSI is a feasible method for KS patients to have a child, and total testicular volume, left testis volume, and neutrophil-to-lymphocyte ratio might be predictors of successful SR.

An examination of predictive markers for successful sperm extraction procedures: a linear model and systematic review.

The authors performed a comprehensive review of current literature to create a model comparing commonly evaluated variables in male factor infertility, for example, follicle-stimulating hormone (FSH), testicular volume (TV), and testosterone (T), to better predict sperm retrieval rate (SRR). Twenty-nine studies were included, 9 with data on conventional testicular sperm extraction (cTESE) for a total of 1227 patients and 20 studies including data on microdissection testicular sperm extraction (mTESE) for a total of 4760 patients. A weighted-means value of SRR, FSH, T, and TV was created, and a weighted linear regression was then used to describe associations among SRR, type of procedure, FSH, T, and TV. In this study, weighted-means values demonstrated mTESE to be superior to cTESE with an SRR of 51.9% vs 40.1%. Multiple weighted linear regressions were created to describe associations among SRR, procedure type, FSH, T, and TV. The models showed that for every 1.19 mIU ml-1 increase in FSH, there would be a significant decrease in SRR by 1.0%. Seeking to create a more clinically relevant model, FSH values were then divided into normal, moderate elevation, and significant elevation categories (FSH <10 mIU ml-1, 10-19 mIU ml-1, and >20 mIU ml-1, respectively). For an index patient undergoing cTESE, the retrieval rates would be 57.1%, 44.3%, and 31.2% for values normal, moderately elevated, and significantly elevated, respectively. In conclusion, in a large meta-analysis, mTESE was shown to be more successful than cTESE for sperm retrievals. FSH has an inverse relationship to SRR in retrieval techniques and can alone be predictive of cTESE SRR.

Evaluation of in vitro fertilization outcome from very low sperm count and microdissection Testicular Sperm Extraction in patients with non-obstructive azoospermia: Paternal age factor

Evaluation of in vitro fertilization outcome from very low sperm count and microdissection Testicular Sperm Extraction in patients with non-obstructive azoospermia: Paternal age factor

Feasibility of repeat microdissection testicular sperm extraction within 6 months for nonobstructive azoospermia.

Microdissection testicular sperm extraction (mTESE) is the mainstay of sperm retrieval in men with nonobstructive azoospermia (NOA). Some experts believe that a resting period of ≥6 months is required between procedures to ensure better sperm retrieval rates and fewer complications. However, no scientific studies have investigated how long patients should wait before arranging a second mTESE procedure. This retrospective study aimed to evaluate whether good success rates are still achieved when a repeat mTESE procedure is performed within 6 months. Total 146 patients with NOA who underwent mTESE twice from the same testis between May 2012 and September 2019 were retrospectively collected. These patients were categorized into three groups according to the time interval between the two mTESE surgeries, with 44, 60, and 42 patients undergoing a repeat mTESE after <3 months (group I), between 3 and 6 months (group II), and >6 months (group III) after the first procedure, respectively. No significant differences were observed between groups in terms of patient characteristics and preoperative hormone profiles. Overall sperm retrieval rates did not differ among the three groups (93.2%, 90.0%, and 88.1% in groups I, II, and III, respectively [ p = 0.719]), nor did fertility outcomes, including rates of fertilization, biochemical pregnancy, clinical pregnancy, and cumulative live births. Sperm retrieval rates for repeat mTESE procedures were consistently high, even when the second procedure was performed within 6 months of the first. Repeat mTESE within 6 months is not a limitation for patients with NOA, if required clinically.

Micro-TESE surgery combined with ICSI regimen in the treatment of non-obstructive azoospermia patients and its effect analysis.

This study aimed to analyze the clinical effects of microdissection testicular sperm extraction (micro-TESE) surgery combined with an intracytoplasmic sperm injection (ICSI) regimen in the treatment of non-obstructive azoospermia (NOA) patients with different etiologies. In total, 128 NOA patients participated in this study, in which they received infertility treatment by micro-TESE surgery combined with an ICSI regimen, and all patients were divided into three groups [the Klinefelter syndrome (KS), the idiopathic and the secondary NOA groups]. In addition, the sperm retrieval rate (SRR), fertilization rate, embryo development status and clinical treatment effects were analyzed. Among the 128 NOA patients, the SRR of KS NOA patients was 48.65%, those of idiopathic and the secondary patients were 33.82% and 73.91%, respectively. Regardless of etiologies, there was no correlation with age, hormone value or testicular volume. Further analysis showed that the SRR of the KS group was positively related with testosterone (T) values, and the SRR of the secondary group had a positive relationship with follicle-stimulating hormone or luteinizing hormone values. In the subsequent clinical treatment, the retrieved sperm was subjected to ICSI and achieved good treatment effects, especially in the secondary group, and the implantation rate (55.56%) and clinical pregnancy rate (68.42%) were both higher than those of the idiopathic group (28.75% and 40.00%) and KS group (22.05% and 30.77%). Micro-TESE surgery combined with ICSI insemination is the most effective treatment regimen for NOA patients. The SRR of NOA patients with different etiologies are related to certain specific factors, and micro-TESE surgery seems to be the ideal and only way to have biological children.