Outcome of repeated micro-surgical testicular sperm extraction in patients with non-obstructive azoospermia

Aim： To evaluate the outcome of repetitive micro-surgical testicular sperm extraction （mTESE） attempts in non-obstructive azoospermia （NOA） cases, in relation to patients＇ initial testicular histology results. Methods： A total of 68 patients with NOA in whom mTESE had been performed in previous intracytoplasmic sperm injection （ICSI） attempts were reviewed. Results： Among the 68 patients with NOA, the first mTESE yielded mature sperm for ICSI in 44 （64%） （Sp^＋）, and failed in the remaining 24 （36%） （Sp^-）. Following their first trial, 24 patients decided to undergo a second mTESE. Of these 24 patients, no spermatozoa were obtained in 5 patients, and Sp^＋ but no fertilization/pregnancy were achieved in 19. In these 24 cases, mTESE was successively repeated for two （n = 24）, three （n = 4） and four （n = 1） times. The second attempt yielded mature sperm in 3/5 patients from the Sp group and 16/19 patients from the Sp^＋ group. At the third and fourth trials, 4/4 and 1/1 of the original Sp^＋ patients were Sp^＋ again, respectively. Distribution of main testicular histology included Sertoli cell-only syndrome （16%）, maturation arrest （22%）, hypospermatogenesis （21%） and focal spermatogenesis （41%）. Overall, in repetitive mTESE, 24/29 （82%） of the attempts were finally Sp^＋. Conclusion： Repeated mTESE in patients with NOA is a feasible option, yielding considerably high sperm recovery rate. In patients with NOA, mTESE may safely be repeated one or more times to increase sperm retrieval rate, as well as to increase the chance of retrieving fresh spermatozoa to enable ICSI.

Microdissection testicular sperm extraction： an update

Patients with non-obstructive azoospermia （NOA） were once considered to be infertile with few treatment options due to the absence of sperm in the ejaculate. In the last two decades, the advent of intracytoplasmic sperm injection （ICSI）, and the application of various testicular sperm retrieval techniques, including fine needle aspiration （FNA）, conventional testicular sperm extraction （TESE） and microdissection testicular sperm extraction （micro-TESE） have revolutionized treatment in this group of men. Because most men with NOA will have isolated regions of spermatogenesis within the testis, studies have illustrated that sperm can be retrieved in most men with NOA, including Klinefelter＇s syndrome （KS）, prior history of chemotherapy and cryptorchidism. Micro-TESE, when compared with conventional TESE has a higher sperm retrieval rate （SRR） with fewer postoperative complications and negative effects on testicular function. In this article, we will compare the efficacy of the different procedures of sperm extraction, discuss the medical treatment and the role of testosterone optimization in men with NOA and describe the micro-TESE surgical technique. Furthermore, we will update our overall experience to allow counseling on the prognosis of sperm retrieval for the specific subsets of NOA.

Seminal plasma anti-Müllerian hormone level correlates with semen parameters but does not predict success of testicular sperm extraction （TESE）

Aim： To assess seminal plasma anti-Müllerian hormone （AMH） level relationships in fertile and infertile males. Methods： Eighty-four male cases were studied and divided into four groups： fertile normozoospermia （n = 16）, oligoastheno- teratozoospermia （n = 15）, obstructive azoospermia （OA） （n = 13） and non-obstructive azoospermia （NOA） （n = 40）. Conventional semen analysis was done for all cases. Testicular biopsy was done with histopathology and fresh tissue examination for testicular sperm extraction （TESE） in NOA cases. NOA group was subdivided according to TESE results into unsuccessful TESE （n = 19） and successful TESE （n = 21）. Seminal plasma AMH was estimated by enzyme linked immunosorbent assay （ELISA） and serum follicular stimulating hormone （FSH） was estimated in NOA cases only by radioimmunoassay （RIA）. Results： Mean seminal AMH was significantly higher in fertile group than in oligoasthenoteratozoospermia with significance （41.5±10.9 pmol/L vs. 30.5±10.3 pmol/L, P 〈 0.05）. Seminal AMH was not detected in any OA patients. Seminal AMH wascorrelated positively with testicular volume （r = 0.329, P = 0.005）, sperm count （r = 0.483, P = 0.007）, sperm motility percent （r = 0.419, P = 0.021） and negatively with sperm abnormal forms percent （r = -0.413, p = 0.023）. Nonsignificant correlation was evident with age （r = -0.155, P = 0.414） and plasma FSH （ r = -0.014, P = 0.943）. In NOA cases, seminal AMH was detectable in 23/40 cases, 14 of them were successful TESE （57.5%） and was undetectable in 17/40 cases, 10 of them were unsuccessful TESE （58.2%）. Conclusion： Seminal plasma AMH is an absolute testicular marker being absent in all OA cases. However, seminal AMH has a poor predictability for successful testicular sperm retrieval in NOA cases.

A laboratory modification to testicular sperm preparation technique improves spermatogenic cell yield

Testicular sperm extraction is a common procedure used to find spermatogenic cells in men with nonobstructive azoospermia. The laboratory processing of biopsied testicular tissues needs to be performed meticulously to acquire a high yield of cells. In this study, the effectiveness of mincing the tissues after testicular biopsy was assessed using histological evaluation, as was the possible adverse effect of residual tissue on the migration of spermatogenic cells during density gradient centrifugation. Our results indicate that testicular residual tissue, when laid on the density gradient medium along with the sperm wash, hinders the spermatogenic cells＇ forming a pellet during centrifugation, and therefore impairs the intracytoplasmic sperm injection procedure. Whereas the mean number of recovered cells from the sperm wash medium （SWM） with residual tissue is 39.435 ~ 24.849, it was notably higher （60.189 ~ 28.214 cells） in the SWM without minced tissues. The remaining tissue contained no functional seminiferous tubules or spermatogenic cells in histological sections. In conclusion, the remaining residual tissue after mincing biopsied testicular tissue does not add any functional or cellular contribution to spermatogenic cell retrieval; in fact, it may block the cellular elements in the accompanying cell suspension from migrating through the gradient layers to form a pellet during centrifugation and cause loss of spermatogenic cells.

Effects of Cryopreservation on the Ultrastructure of Human Testicular Sperm

Objective To investigate effects of cryopreservation on changes of the ultrastructure of human testicular sperm and evaluate the efficacy of cryopreserving testicular tissue as a source of sperm for assisted reproduction. Methods Testicular biopsy tissues were obtained from infertile patients （n=12） with obstructive azoospermia and cryopreserved. Testicular sperm motility was observed after in vitro culture procedure. The ultrastructure of testicular sperm （n=6） was examined by transmission electron microscope. Results After cryopreservation, 10 biopsy tissues frozen revealed motile sperm, and 2 samples showed non-motile sperm. Some testicular sperm in frozen-thawed group had normal morphology in fine structures. Sperm head in frozen-thawed tissue showed a proportion of nuclei with more electron-dense granules of chromatin. In a few frozen-thawed sperm heads, formation of vesicles and degeneration were observed. The frozen-thawed testicular sperm frequently showed swollen or/and ruptured of the plasma membrane and acrosome membranes. Conclusion Cryopreservation of testicular tissue is simple and efficacious for testicular sperm extraction. And the freezing-thawing procedure of testicular tissue causes damage to ultrastructural morphology of human testicular sperm.

Fertilization of in vitro matured human oocytes by intracytoplasmic sperm injection (ICSI) using ejaculated and testicular spermatozoa

Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.

Probability to retrieve testicular spermatozoa in azoospermic patients

Aim: The degree of probability to retrieve spermatozoa from testicular tissue for intracytoplasmic sperm injection intooocytes is of interest for counselling of infertility patients. We investigated the relation of sperm retrieval to clinical dataand histological pattern in testicular biopsies from azoospermic patients. Methods: In 264 testicular biopsies from142 azoospermic patients, the testicular tissue was shredded to separate the spermatozoa, histological semi-thin sec-tions of which were then evaluated using Johnsen score. Results: The retrieval of spermatozoa correlated signifi-cantly (P < 0.001) with the testicular volume (r = 0.49), the FSH concentration ( r = -0.66), the maximum score(r = 0.85) and the mean Johnsen score (r = 0.81). In the multivariate regression analysis the successful testicularsperm extraction showed the closest relationship to the maximum score. The testicular volume correlated significantlywith the mean Johnsen score ( r = 0.64, P < 0. 001), and the basal serum FSH concentration mainly with the maxi-mum score ( r = -0.77; P < 0. 001 ). Patients with a history cryptorchidism showed a significantly lower Johnsenscore compared to the patients who did not have any testicular disease in the past (3.7 ± 2.4 vs. 5.9 ± 2. 5; P < O.01). Conclusion: In a limited range, the testicular volume and the FSH concentration in serum were related to theJohnsen score which correlated significantly with the sperm retrieval. The successful sperm retrieval can be expected inall azoospermic patients irrespective of the results of clinical examination. However, the probability of retrieval of sper-matozoa decreased significantly in patients with a FSH level > 18 U/L, tesficular volume < 5mL, mean Johnsen score< 5, and maximum Johnsen score < 7.

Development of a predictive model for increasing sperm retrieval sucpess 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-Miillerian 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-Mullerian hormone(odds ratio[OR]=0.902,95%confidence interval[Cl]:0.821-0.990),inhibin B(OR=1.012,95%Cl:1.001-1.024),Klinefelter’s syndrome(OR=0.022,95%Cl:0.002-0.243),Y chromosome microdeletion(OR=0.050,95%Cl:0.005-0.504),cryptorchidism with orchiopexy(OR=0.085,95%Cl:0.008-0.929),and idiopathic nonobstructive azoospermia(OR=0.031,95%Cl:0.003-0.277).The prediction model had an area under the curve of 0.720(95%Cl: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.

Testicular sperm extraction(TESE)outcomes in the context of malignant disease:a systematic review

Advances in the oncology field have led to improved survival rates.Consequently,quality of life after remission is anticipated,which includes the possibility to conceive children.Since cancer treatments are potentially gonadotoxic,fertility preservation must be proposed.Male fertility preservation is mainly based on ejaculated sperm cryopreservation.When this is not possible,testicular sperm extraction(TESE)may be planned.To identify situations in which TESE has been beneficial,a systematic review was conducted.The search was carried out on the PubMed,Scopus,Google Scholar,and CISMeF databases from 1 January 2000 to 19 March 2020.Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA)recommendations were followed in selecting items of interest.Thirty-four articles were included in the systematic review,including 15 articles on oncological testicular sperm extraction(oncoTESE),18 articles on postgonadotoxic treatment TESE and 1 article on both oncoTESE and postgonadotoxic treatment TESE.Testicular sperm freezing was possible for 42.9%to 57.7%of patients before gonadotoxic treatment and for 32.4%to 75.5%of patients after gonadotoxic treatment,depending on the type of malignant disease.Although no formal conclusion could be drawn about the chances to obtain sperm in specific situations,our results suggest that TESE can be proposed before and after gonadotoxic treatment.Before treatment,TESE is more often proposed for men with testicular cancer presenting with azoospermia since TESE can be performed simultaneously with tumor removal or orchiectomy.After chemotherapy,TESE may be planned if the patient presents with persistent azoospermia.

Combination of serum inhibin B and follicle-stimulating hormone levels can not improve the diagnostic accuracy on testicular sperm extraction outcomes in Chinese non-obstructive azoospermic men

Background It is still controversial whether the serum inhibin B level is a superior predictor of the presence of sperm in testicular sperm extraction （TESE） in azoospermic men compared with serum follicle-stimulating hormone （FSH）. In this study, we evaluated the diagnostic accuracy of serum inhibin B levels as a predictor of the outcome of TESE in Chinese non-obstructive azoospermic men and compared it with the traditional marker serum FSH and testicular volumes. Methods Basal values of serum hormone levels, testicular volumes and histological evaluation of 305 Chinese non-obstructive azoospermic men were analyzed. The level of inhibin B was measured using a three-step enzyme-linked immunoassay before sperm extraction, and the diagnostic accuracy of prediction of the outcome of TESE was compared for different markers by the receiver operating characteristics （ROC） curve analysis. Results Testicular sperm was successfully retrieved in 137 of 305 patients （44.9%）. The serum level of inhibin B, the FSH and the testicular volume were significantly different between the successful TESE group and the unsuccessful group. According to the ROC curve analysis, for inhibin B, the cut-off value for discriminating between successful and failed TESE was 28.39 pg/ml （sensitivity 83.5%, specificity 79.1%）. For FSH, the best cut-off value for discriminating was 11.05 pg/ml （sensitivity 83.5%, specificity 74.5%）. The area under the ROC curve of serum inhibin B was similar to that of FSH. Combining the serum inhibin B with FSH levels did not improve the predictive value for successful TESE. Conclusions Serum inhibin B and FSH levels are correlated with spermatogenesis. However, inhibin B is not superior to FSH in predicting the presence of sperm in TESE. And the combination of them does not improve the diagnostic accuracy on TESE outcome.

Use of testicular sperm for intracytoplasmic sperm injection in men with high sperm DNA fragmentation： a SWOT analysis

Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation （SDF） in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology （ART）, an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection （Testi-ICSI）. In this article, we used a SWOT （strengths, weaknesses, opportunities, and threats） analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval.

Testicular sperm extraction and intracytoplasmic sperm injection in non obstructive azoospermia

Objective To report the experience in sperm extraction from testicular biopsies (TESE) performed from March 1996 to July 1998 in men with non obstructive azoospermia (NOA) Methods Comparisons of age, volume of both testes, serum FSH and testosterone in men, and histology of testicular samples in the first cycles between cycles with spermatozoa found and those without spermatozoa found were performed Comparisons of fertilization, cleavage and pregnancy rates between cycles with spermatozoa injected and those with spermatids injected were performed Results Spermatozoa were found in only 12 out of 26 first TESE cycles (46 2%) and other cycles had spermatids (round cells) only Age of men, history of mumps orchitis/oligozoospermia, volume of both testes and serum FSH/testosterone levels in men were not significantly different between cycles with and without spermatozoa The fertilization rate was significantly higher in cycles with spermatozoa injected than those with round cell injections (63 3% vs 23 2%, P <0 0001, Chi squared test) The pregnancy rate was 14 3% per cycle when spermatozoa were injected Conclusions TESE followed by Intracytoplasmic sperm injection (ICSI) is an effective treatment in patients with NOA Less than half of the patients undergoing TESE had spermatozoa recovered Age of men, volume of both testes and serum FSH/ testosterone levels in men were not useful in predicting successful recovery Compared to using ejaculated and epididymal spermatozoa, fertilization and pregnancy rates were achieved when testicular spermatozoa were used for ICSI

Sperm retrieval rates and clinical outcomes for patients with different causes of azoospermia who undergo microdissection testicular sperm extraction-intracytoplasmic sperm injection

The aim of our study was to compare the sperm retrieval rates(SRRs)and clinical outcomes of patients with different causes of azoospermia who underwent microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI).We conducted a retrospective study at the Reproductive Medicine Center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.This study examined 769 patients with nonobstructive azoospermia who underwent 347 cycles of micro-TESE-ICSI.Patients with azoospermia were classified into Group A(Klinefelter syndrome,n=284,125 cycles),Group B(azoospermia Y chromosome factor c[AZFc]microdeletion,n=91,64 cycles),Group C(cryptorchidism,n=52,39 cycles),Group D(previous mumps and bilateral orchitis,n=23,23 cycles),and Group E(idiopathic azoospermia,n=319,96 cycles).Clinical characteristics,SRR,embryonic development,and pregnancy outcomes of the patients were compared between all groups.Patients in Group D had the highest and most successful SRR.The average SRR for all patients was 46.0%.The rates of clinical pregnancy,implantation,and live birth in Group D were 78.3%,65.0%,and 74.0%,respectively,which were higher than those in all other groups(P<0.05).Group B patients had the lowest clinical pregnancy,implantation,and live birth rates of all groups(P<0.05).No differences were found in the miscarriage rate or birth defects among the groups(P>0.05).Patients with orchitis had the highest SRR and best clinical outcomes.Although AZFc microdeletion patients had a higher SRR,their clinical outcomes were worse.

Use of testicular sperm in couples with SCSA-defined high sperm DNA fragmentation and failed intracytoplasmic sperm injection using ejaculated sperm

Sperm DNA fragmentation(SDF)has been linked with male infertility,and previous studies suggest that SDF can have negative influence on pregnancy outcomes with assisted reproduction.We performed a retrospective review of consecutive couples with a high SDF level that had intracytoplasmic sperm injection(ICSI)using testicular sperm(T-ICSI).We compared the T-ICSI outcomes to that of two control groups:87 couples with failed first ICSI cycle and who had a second ICSI cycle using ejaculated sperm(Ej-ICSI),and 48 consecutive couples with high sperm chromatin structure assay(SCSA)-defined SDF(>15%)that underwent an ICSI cycle using ejaculated sperm after one or more failed ICSI cycles(Ej-ICSI-high SDF).The mean number of oocytes that were retrieved and the total number of embryos were not different among the three groups.The mean number of transferred embryos in the T-ICSI group was higher than the Ej-ICSI group but not significantly different than the Ej-ICSI-high SDF group(1.4,1.2,and 1.3,respectively,P<0.05).Clinical pregnancy rate in the T-ICSI group was not significantly different than the Ej-ICSI and Ej-ICSI-high SDF groups(48.6%,48.2%,and 38.7%,respectively,P>0.05).No significant difference was found in live birth rate when comparing T-ICSI to Ej-ICSI and Ej-ICSI-high SDF groups.The results suggest that pregnancy outcomes and live birth rates with T-ICSI are not significantly superior to Ej-ICSI in patients with an elevated SCSA-defined sperm DNA fragmentation and prior ICSI failure(s).

Reproductive outcomes of intracytoplasmic sperm injection using testicular sperm and ejaculated sperm in patients with AZFc microdeletions:a systematic review and meta-analysis

Studies have explored the assisted reproductive technology(ART)outcomes of Y-chromosome azoospermia factor c(AZFc)microdeletions,but the effect of sperm source on intracytoplasmic sperm injection(ICSI)remains unknown.To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions,we searched Embase,Web of Science,and PubMed to conduct a systematic review and meta-analysis.The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group(risk ratio:0.97,95%confidence interval[CI]:0.73-1.28,P=0.82).The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group(risk ratio:1.06,95%Cl:0.54-2.06,P=0.87).The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group(risk ratio:1.24,95%Cl:0.66-2.34,P=0.50).Inevitable heterogeneity weakened our results.However,our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes,representing a meaningful result for clinical treatment.More properly designed studies are needed to further confirm our conclusions.

Clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection with fresh or cryopreserved sperm in patients with nonobstructive azoospermia

We performed this study to evaluate the clinical outcomes of microdissection testicular sperm extraction-intracytoplasmic sperm injection(micro-TESE-ICSI)treatment that used fresh or cryopreserved sperm in patients with nonobstructive azoospermia(NOA).A total of 338 NOA patients with 344 consecutive cycles received treatment in the reproductive medicine center of Peking University Third Hospital in Beijing,China,from January 2014 to December 2017.Fresh oocytes and fresh sperm were used in 222 patients with 234 cycles(Group A).Fresh oocytes and cryopreserved sperm were used in 116 patients with 110 cycles(Group B).We compared patient characteristics,embryonic development,and pregnancy outcomes between Groups A and B.There was no statistical difference in the patient characteristics,and no differences were observed with fertilization or quality embryo rates between Groups A and B.The rates of clinical pregnancy and live birth were both higher for Group A than those for Group B(both P<0.05).In conclusion,fresh testicular sperm appears to produce better ICSI outcomes than cryopreserved testicular sperm in patients with NOA.

Predictive factors for successful sperm retrieval by microdissection testicular sperm extraction in men with nonobstructive azoospermia and a history of cryptorchidism

This study aims to explore the factors influencing the success rate of the microdissection testicular sperm extraction(Micro-TESE)in patients with nonobstructive azoospermia(NOA)and cryptorchidism.Clinical data of 162 patients with cryptorchidism who underwent Micro-TESE due to infertility from December 2015 to May 2020 in the First Affiliated Hospital of Nanjing Medical University were analyzed retrospectively.In the univariate analysis,significant differences in the age of patient at the time of orchidopexy(median[interquartile range,IQR]:7.0[4.0–11.0]years vs 11.5[9.0–14.5]years,P<0.001),interval between orchidopexy and Micro-TESE(mean±standard deviation:17.5±5.0 years vs 14.4±4.4 years,P<0.001),severity of cryptorchidism(unilateral[62.8%]vs bilateral[31.6%],P<0.001;location of cryptorchidism,intra-abdominal[27.3%]vs inguinal[44.8%]vs suprascrotal[66.7%],P<0.001),volume of the dominant testis(median[IQR]:17.00[15.00–19.00]ml vs 14.50[11.75–16.25]ml,P<0.001),and levels of follicle-stimulating hormone(FSH;P=0.004)and testosterone(P=0.006)were observed between the successful and failed sperm extraction groups.After conducting the multivariate analysis,four of these factors,including unilateral/bilateral cryptorchidism(P<0.001),location of cryptorchidism(P=0.032),age of orchidopexy(P<0.001),and dominant testicular volume,were adopted in the clinical prediction model to evaluate preoperatively the success rate of Micro-TESE for patients with NOA and cryptorchidism.The likelihood of successful sperm retrieval by Micro-TESE in men with NOA and cryptorchidism increased in patients with mild forms of cryptorchidism.

Follow-up of children born after intracytoplasmic sperm injection with epididymal and testicular spermatozoa

Background To evaluate the safety of intracytoplasmic sperm injection （ICSI） with epididymal or testicular sperm, this study compared children born after ICSI treatment with epididymal or testicular sperm with children conceived after ICSI with ejaculated sperm. Methods This retrospective study included 317 children born after ICSI with percutaneous epididymal sperm aspiration （PESA）, 103 children born after ICSI with testicular sperm aspiration （TESA）, and a control group of 1008 children born after ICSI with ejaculated sperm. All of the patients received their assisted reproductive treatment in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2004 to December 2011. Data, such as the rate of stillbirths, perinatal mortality, gestational age, birth weight, and the rate of congenital malformations of the three groups, were compared. Results PESA and TESA children were not different from ICSI children in the rate of stillbirths, perinatal mortality, infant mortality rate, gestational age, the rate of prematurity, and the rate of malformations （P〉0.05）. A slight increase in birth defects was reported in the TESA group compared with those in the control group, but there was no significant difference between the groups （P〉0.05）. Conclusion ICSI with epididymal or testicutar sperm does not lead to more stillbirths or congenital malformations compared with ICSI using ejaculated sperm.

A preliminary study of sperm identification in microdissection testicular sperm extraction samples with deep convolutional neural networks

Sperm identification and selection is an essential task when processing human testicular samples for in vitro fertilization.Locating and identifying sperm cell(s)in human testicular biopsy samples is labor intensive and time consuming.We developed a new computer-aided sperm analysis(CASA)system,which utilizes deep learning for near human-level performance on testicular sperm extraction(TESE),trained on a custom dataset.The system automates the identification of sperm in testicular biopsy samples.A dataset of 702 de-identified images from testicular biopsy samples of 30 patients was collected.Each image was normalized and passed through glare filters and diffraction correction.The data were split 80%,10%,and 10%into training,validation,and test sets,respectively.Then,a deep object detection network,composed of a feature extraction network and object detection network,was trained on this dataset.The model was benchmarked against embryologists’performance on the detection task.Our deep learning CASA system achieved a mean average precision(mAP)of 0.741,with an average recall(AR)of 0.376 on our dataset.Our proposed method can work in real time;its speed is effectively limited only by the imaging speed of the microscope.Our results indicate that deep learning-based technologies can improve the efficiency of finding sperm in testicular biopsy samples.

Laboratory and Clinical Outcomes of Single Sperm Cryopreservation in Patients Underwent Micro-TESE

For men with severe oligozoospermia, sperm cryopreservation can preserve surgically obtained sperm. How to cryopreserve single sperm in men is still a hot topic in assisted reproduction technology. Aim to analyze the laboratory and pregnancy outcomes of single sperm cryopreservation group, we retrospectively selected 38 cycles underwent single sperm cryopreservation and thawing as the study group and 618 cycles underwent conventional sperm cryopreservation and thawing as the control group, which were performed in the reproductive medicine center of the Sixth Affiliated Hospital, Sun Yatsen University, from April 2014 to October 2023. All the sperm came from microdissection testicular sperm extraction (micro-TESE), and performed intracytoplasmic sperm injection (ICSI) for fertilization. Zygotes were cultured to Day 3 embryo, which were freshly transferred to female uterus. Surplus embryos were cultured to blastosphere and cryopreserved. There was no statistical difference in female/male age, female BMI, infertility duration and female basal sex hormone (FSH, LH E2, AMH), No. of oocytes retrieved per cycle, No. of ICSI oocytes per cycle and No. of embryos transferred per cycle between the two groups (P > 0.05). No significant difference was found in two-pronuclear oocyte fertilization rate (59.23% VS 58.84%), Day 3 available embryo rate (61.81% VS 63.55%), Day 3 good-quality embryo rate (45.73% VS 50.27%), blastocyst formation rate (47.83% VS 49.46%), the implantation rate (47.37% VS 52.16%), clinical pregnancy rate (36.84% VS 47.18%), miscarriage rate (14.29% VS 12.68%) and live birth rate (85.71% VS 81.70%) between two groups (P > 0.05). In conclusion, single-sperm cryopreservation was the optimal method to preserve sperm after micro-TESE. It can increase the utilization of each sperm and lead to clinical pregnancy.