Surgical recovery of sperm in non-obstructive azoospermia

The development of intracytoplasmic sperm injection (ICSI) opened a new era in the field of assisted reproduction and revolutionized the assisted reproductive technology protocols for couples with male factor infertility. Fertilisation and pregnancies can be achieved with spermatozoa recovered not only from the ejaculate but also from the seminiferous tubules. The most common methods for retrieving testicular sperm in non-obstructive azoospermia (NOA) are testicular sperm aspiration (TESA: needle/fine needle aspiration) and open testicular biopsy (testicular sperm extraction: TESE). The optimal technique for sperm extraction should be minimally invasive and avoid destruction of testicular function, without compromising the chance to retrieve adequate numbers of spermatozoa to perform ICSI. Microdissection TESE (micro-TESE), performed with an operative microscope, is widely considered to be the best method for sperm retrieval in NOA, as larger and opaque tubules, presumably with active spermatogenesis, can be directly identified, resulting in higher spermatozoa retrieval rates with minimal tissue loss and low postoperative complications. Micro-TESE, in combination with ICSI, is applicable in all cases of NOA, including Klinefelter syndrome (KS). The outcomes of surgical sperm retrieval, primarily in NOA patients with elevated serum follicle-stimulating hormone (FSH) (NOA including KS patients), are reviewed along with the phenotypic features. The predictive factors for surgical sperm retrieval and outcomes of treatment were analysed. Finally, the short- and long-term complications in micro-TESE in both 46XY males with NOA and KS patients are considered.     

Male factors determining the outcome of intracytoplasmic sperm injection with epididymal and testicular spermatozoa

During a period of 8 years, 1,079 intracytoplasmic sperm injection (ICSI) procedures with aspirated epididymal or testicular spermatozoa were performed. Epididymal spermatozoa were used in 172 cycles and testicular spermatozoa or spermatids in 907 cycles. Multiple biopsies were obtained from at least two different locations in the testes. Retrieved spermatozoa were used after cryopreservation (frozen) or immediately after aspiration (fresh). Three hundred patients had obstructive azoospermia (OA) or ejaculation failure. In 414 cases, azoospermia was caused by impaired spermatogenesis resulting from maldescended testes, chemotherapy/radiotherapy, or by Sertoli-cell-only syndrome, genetic disorders or unknown aetiology. Transfer rates, pregnancy rates and birth rates per ICSI cycle showed no statistically significant differences between testicular and epididymal spermatozoa in men with OA (28% average birth rates in both cases). However, birth rates differed significantly with regard to the status of spermatogenesis. Treatment of men with nonobstructive azoospermia (NOA) resulted in a birth rate of 19% per cycle. In all patient groups, there was no difference in the birth rates achieved with fresh and cryopreserved spermatozoa. While testicular volume, follicle-stimulating hormone level and age of the male patient are no statistically significant prognostic factors, the underlying cause of azoospermia is the most important factor determining the outcome of ICSI with epididymal and testicular spermatozoa. The pregnancy rate is lower in NOA patients than in those with OA.     

Effects of obesity on sperm retrieval,early embryo quality and clinical outcomes in men with nonobstructive azoospermia undergoing testicular sperm aspiration‐intracytoplasmic sperm injection cycles

The objective of this study was to assess the effects of body mass index (BMI) on sperm retrieval, early embryo quality and clinical outcomes in patients with nonobstructive azoospermia (NOA) undergoing testicular sperm aspiration‐intracytoplasmic sperm injection (TESA‐ICSI). A total of 3,005 infertile couples were evaluated between January 2010 and June 2017, including 1585 normal‐weight (BMI < 25 kg/m²), 847 overweight (BMI 25–29.99 kg/m²) and 573 obese (BMI ≥ 30 kg/m²) patients. We found no significant relationship between BMI and sperm retrieval rate (22.4%, 24.3% and 25.1%, p = 0.327) or sperm motility. Among the 705 patients with NOA who underwent TESA‐ICSI cycles, obese individuals had lower T levels and higher E2 levels than normal‐weight and overweight individuals. However, there were no significant differences in other male hormones (follicle stimulating hormone [FSH], luteinizing hormone [LH], or prolactin [PRL]) among the groups. We also found that the sperm parameters, embryo quality and clinical outcomes of patients with NOA undergoing TESA‐ICSI were not influenced by high BMI levels. In conclusion, this study demonstrated a lack of obvious effects of obesity on sperm retrieval, early embryo quality and clinical outcomes in infertile men undergoing TESA‐ICSI cycles, although T and E2 levels were affected.     

Intracytoplasmic sperm injection by testicular sperm in patients with aspermia or azoospermia after cancer treatment

The aim of this retrospective study was to evaluate the efficiency of testicular biopsy and intracytoplasmic sperm injection (ICSI) in patients with aspermia or non-obstructive azoospermia (NOA) after cancer treatment. From 1996 to 2003, 30 men with a history of cancer, affected by aspermia or NOA and without sperm cryopreserved before cytotoxic treatment underwent testicular sperm extraction (TESE). In these men, clinical, hormonal and histological characteristics were compared; 13 underwent 39 TESE-ICSI cycles using frozen-thawed testicular spermatozoa (TESE-ICSI group). In the same period, 31 ICSI cycles were performed in 20 men with aspermia or NOA using ejaculated sperm frozen before cancer treatment (ejaculated sperm-ICSI group). Fertilization, blastocyst development, pregnancy and miscarriage rates were compared between the groups. Testicular volume, serum follicle-stimulating hormone level and Johnsen score indicated complete although reduced spermatogenesis in men with aspermia and abnormal spermatogenesis in men with NOA. After TESE, sperm retrieval was positive in 92% of men with aspermia and 58% of men with NOA. In TESE-ICSI patients with NOA a significantly lower proportion of embryos developed to the blastocyst stage than in patients with aspermia and in those after ICSI with frozen-thawed ejaculated sperm (23% vs. 43% and 47%, p = 0.03 and p < 0.01 respectively). In all groups the miscarriage rates were high; in patients with aspermia and NOA, characterized by increased age, the miscarriage rate tended to be higher in spite of similar female age and female indications of infertility. In patients affected by aspermia or NOA after cancer treatment and without sperm cryopreserved before treatment, TESE-ICSI using testicular sperm provide a chance to father a child.     

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.     

Efficiency of percutaneous testicular sperm aspiration as a mode of sperm collection for intracytoplasmic sperm injection in nonobstructive azoospermia

PURPOSE: We determined the feasibility of obtaining mature spermatozoa for intracytoplasmic sperm injection (ICSI) by percutaneous testicular sperm aspiration in men with nonobstructive azoospermia. We also compared the results of ICSI using spermatozoa recovered by open excisional biopsy versus percutaneous testicular sperm aspiration. MATERIALS AND METHODS: A total of 84 men with nonobstructive azoospermia underwent percutaneous testicular sperm aspiration to recover testicular spermatozoa for ICSI on the day of ova retrieval from the wife. Percutaneous testicular sperm aspiration was performed with the patient under general anesthesia in the upper and lower poles of each testis. It was followed by immediate microscopic search of the aspirate to confirm the presence of spermatozoa. In the absence of spermatozoa open excisional biopsy was performed in the same setting. RESULTS: Percutaneous testicular sperm aspiration resulted in the recovery of mature spermatozoa in 45 men (53.6%). Of the remaining 39 men (46.4%) requiring open biopsy adequate spermatozoa were recovered in 28 (71.8%). Although the fertilization rate was significantly higher in the sperm aspiration group, the cleavage and pregnancy rates were similar in the 2 groups. CONCLUSIONS: Percutaneous testicular sperm aspiration was a successful initial approach to collect mature spermatozoa in a high proportion of men with nonobstructive azoospermia. It is safe, minimally invasive and well tolerated by all patients.     

冻融复苏微量精子行卵细胞胞质内单精子注射术的疗效及临床妊娠结局分析

目的:回顾性分析123例无精子症患者经皮附睾精子抽吸术(PESA)或经皮睾丸精子抽吸术(TESA)后冻融复苏微量精子行卵细胞胞质内单精子注射术(ICSI)的疗效及临床妊娠结局情况。方法:将采用微量冻融PESA、TESA精子行ICSI的病例归为冻融精子组,采用新鲜PESA、TESA精子行ICSI的病例归为对照组。比较冻融精子组与新鲜精子组组间及组内的双原核(2PN)受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率、多胎妊娠率有无统计学差异。结果:PESA精子冻融组与新鲜组受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率及多胎妊娠率分别为75.67%vs76.49%,64.96%vs66.19%,55.21%vs57.22%,13.21%vs12.61%,3.77%vs5.41%,37.74%vs37.84%(P>0.05),TESA精子冻融组与新鲜组受精率、优质胚胎率、临床妊娠率、流产率、宫外孕率及多胎妊娠率分别为74.41%vs76.43%,64.63%vs66.35%,46.81%vs53.39%,18.18%vs14.55%,4.55%vs1.82%,37.74%vs37.84%,组间及组内均无统计学差异(P>0.05)。PESA精子与TESA精子冻融复苏成功率为70.07%vs62.67%,无统计学差异(P>0.05)。结论:微量PESA及TESA精子冻融技术对无精子症患者来说是一种安全、经济、有效的治疗方法;精子冷冻复苏技术有待于进一步提高;该技术是否会增加子代远期遗传风险仍有待于进一步探讨和研究。     

ICSI outcome in patients with high DNA fragmentation: Testicular versus ejaculated spermatozoa

Sperm DNA fragmentation (SDF) has emerged as an important biomarker in the assessment of male fertility potential with contradictory results regarding its effect on ICSI. The aim of this study was to evaluate intracytoplasmic sperm injection (ICSI) outcomes in male patients with high SDF using testicular versus ejaculated spermatozoa. This is a prospective study on 36 men with high‐SDF levels who had a previous ICSI cycle from their ejaculates. A subsequent ICSI cycle was performed using spermatozoa retrieved through testicular sperm aspiration. Results of the prior ejaculate ICSI were compared with those of the TESA‐ICSI. The mean (SD) SDF level was 56.36% (15.3%). Overall, there was no difference in the fertilization rate and embryo grading using ejaculate and testicular spermatozoa (46.4% vs. 47.8%, 50.2% vs. 53.4% respectively). However, clinical pregnancy was significantly higher in TESA group compared to ejaculated group (38.89% [14 of 36] vs. 13.8% [five of 36]). Moreover, 17 live births were documented in TESA group, and only three live births were documented in ejaculate group (p < .0001). We concluded that the use of testicular spermatozoa for ICSI significantly increases clinical pregnancy rate as well as live‐birth rate in patients with high SDF.     

Testicular versus ejaculated spermatozoa in ICSI cycles of normozoospermic men with high sperm DNA fragmentation and previous ART failures

As a part of male assessment, conventional sperm parameters including morphologic features have been dedicated as major factors influencing fertilisation and pregnancy rates in assisted reproductive technology (ART). Genomic integrity of spermatozoa has also been found to influence fertility prognosis, and hence, sperm DNA fragmentation index (DFI) has been adopted by many centres to document this entity. Despite several suggested approaches, there is lack of universal consensus on optimising fertility outcomes in males with high sperm DFI. In this context, the results from cycles using testicular spermatozoa (TESA) obtained by aspiration were compared with those of ejaculated spermatozoa (EJ) in normozoospermic subjects with high sperm DFI and previous ART failures. Clinical (41.9% versus 20%) and ongoing pregnancy rates (38.7% versus 15%) were significantly better and miscarriages were lower in TESA group when compared to EJ group. Sperm DFI should be a part of male partner's evaluation following unsuccessful ART attempts. When high DFI is detected (>30%), ICSI using testicular spermatozoa obtained by TESA seems an effective option particularly for those with repeated ART failures in terms of clinical, ongoing pregnancies and miscarriages even though conventional sperm parameters are within normal range.     

Clinical use of pentoxifylline for activation of immotile testicular sperm before ICSI in patients with azoospermia

The testicular sperm from biopsy and frozen/thawed tissue are frequently immotile. The purpose of our retrospective study was to assess the effect of short exposure of testicular samples with only immotile sperm to pentoxifylline (PF)-sperm motility stimulator. In 77 of 294 (26.2%) testicular sperm ablation/testicular sperm extraction-intracytoplasmic sperm injection (TESA/TESE-ICSI) cycles in patients with azoospermia, only immotile sperm were found in biopsies even after 2 hours of incubation of tissue in the medium. These 77 cycles were divided into 2 groups. In group 1 (cycles between 1999 and 2001; n = 30), ICSI was performed with untreated immotile sperm. In group 2 (cycles between 2002 and 2004; n = 47), immotile testicular sperm were treated for 20 minutes with pentoxifylline (PF) (1.76 mM) before ICSI. Both groups had the same proportion of ICSI cycles with fresh, frozen/thawed, and aspirated testicular sperm. The overall pregnancy rate of TESA/TESE-ICSI did not vary during the study period. In 45 of 47 (95.7%) testicular samples with total immotility, the sperm started to move 20 minutes after PF treatment. The mean time required for ICSI was shortened in the PF group (30 minutes [minimum 10, maximum 90] vs 120 minutes [minimum 60, maximum 240]) due to easier identification of motile sperm. In comparison with the nontreated group, the PF group had a higher fertilization rate (66% vs 50.9%; P < .005) and mean number of embryos per cycle (4.7 +/- 3.3 vs 2.7 +/- 2.1; P < .01). The clinical pregnancy rate per cycle in PF and non-PF groups was 38.3% and 26.7%, respectively. By using PF in cases of only immotile testicular sperm we can cause movement of testicular sperm, allow easier identification of vital sperm, shorten the procedure, improve fertilization rates, and increase the number of embryos.     

Comparison of pregnancy and neonatal outcomes of intracytoplasmic sperm injection performed with frozen versus fresh testicular sperm

BackgroundIt remains controversial whether there is a difference in the prognosis of intracytoplasmic sperm injection (ICSI) using frozen or fresh testicular sperm in patients with obstructive azoospermia (OA). Moreover, in the available studies, few have tracked neonatal outcomes. This study aimed to compare the pregnancy and neonatal outcomes of ICSI using cryopreserved sperm versus fresh sperm collected by testicular sperm aspiration (TESA).MethodsA total of 317 OA patients treated with ICSI in a university affiliated hospital from January 2016 to December 2020 were included in this study. The participants were divided into two groups according to the type of sperm used for ICSI: frozen sperm group (n=154) and fresh sperm group (n=163). The pregnancy and neonatal outcomes of the two groups were compared.ResultsThe data produced by this study showed no significant statistical difference in the 2 pronuclei (2PN) fertilization rate, 2PN cleavage rate, high-quality blastocyst rate, and the average number of transferred embryos in the frozen and fresh sperm groups. Similarly, no difference was found in implantation rate, clinical pregnancy rate, multiple pregnancy rate, miscarriage rate, premature delivery rate, live birth rate, and gender ratio at birth (P>0.05). The average newborn birth weight was similar in both groups (2,932.61±728.40 vs. 3,100.32±515.64 g, respectively) (P>0.05). A higher incidence of low birthweight (LBW) newborns was found in the frozen sperm group (20.91% vs. 8.49%) (P<0.05). Multiple logistic regression analysis showed that LBW is related to single or twin pregnancies (P<0.01), but not sperm (frozen or fresh) (P>0.05). We further analyzed the twin and single pregnancies in the two groups separately, and found that the incidences of LBW were both similar (P>0.05). There was no difference in the Apgar scores at 1 min and 5 min after birth between the two groups (P>0.05).ConclusionsThe use of frozen testicular sperm by TESA was efficient for men with OA. There were similar pregnancy and neonatal outcomes following TESA-ICSI using frozen or fresh sperm in this retrospective study. Prospective investigations are needed for further validation.     

卵子玻璃化冷冻在睾丸取精失败周期中的临床应用(附8例报告)

目的:探讨卵子玻璃化冷冻在睾丸取精失败周期中的临床应用价值。方法:回顾分析体外受精周期取卵日因男方无精子症行睾丸取精失败且无供精接受卵子冷冻的8例不孕症妇女。所有卵子经玻璃化冷冻保存2个月后解冻,存活的成熟卵子采用与其丈夫血型一致的精子库冷冻精子行卵细胞胞质内单精子注射(ICSI)受精,记录卵子的存活、受精、卵裂、优质胚胎、临床妊娠及分娩情况。结果:冷冻-解冻60枚卵子中存活47枚,存活率为78.3%(47/60);其中41枚成熟卵子行ICSI受精,正常受精率为80.5%(33/41),卵裂率为81.8%(27/33),优质胚胎率为59.3%(16/27);15枚胚胎移植给8例患者,移植周期率为100%(8/8),平均移植胚胎数为1.9±0.8。B超显示有5个孕囊形成,胚胎种植率为33.3%(5/15);5例患者获得了临床妊娠,临床妊娠率为62.5%(5/8),均为宫内单活胎。4例已顺利分娩3男婴1女婴,体重为(3 787.5±513.7)g,染色体及发育均正常。结论:卵子玻璃化冷冻是睾丸取精失败患者的有效保障,与精子库冷冻供精受精可以获得理想临床结局。     

非梗阻性无精子症患者睾丸穿刺获精子结局的预测因素研究

目的探讨非梗阻性无精子症(NOA)患者睾丸体积、血清抑制素B(INHB)及性激素水平对预测睾丸穿刺(TESA)获精子结局的意义。方法实验组为162例NOA患者,按TESA获精子结局分为有精子组(TESA^+,n=74)和无精子组(TESA^-,n=88);正常组为60例同期精液常规参数正常者。比较各组的睾丸体积、血清INHB及性激素水平,筛选有显著性差异的指标,应用ROC曲线评价其对预测TESA获精子结局的意义。结果睾丸体积、血清INHB及FSH三项指标在TESA^+组和TESA^-组间存在显著性差异(P<0.05),因此为优选的预测指标。三者的ROC曲线下面积(AUC)分别为0.816、0.861、0.777,且后两者间有显著性差异(P<0.05),最佳切点值分别为8.15 ml、70.15 pg/ml、5.52 U/L,对应的诊断灵敏度分别为93.2%、87.8%、74.3%,特异度分别为65.9%、81.8%、77.3%。结论睾丸体积、血清INHB及FSH对预测NOA患者TESA获精子结局有重要意义,且INHB的预测意义优于FSH。各指标的预测意义与其对应的诊断灵敏度和特异度不成正相关,表明联合诊断的重要性。     

Analysis of the outcome of intracytoplasmic sperm injection using fresh or frozen sperm

Study Type – Diagnostic (retrospective cohort)
Level of Evidence 2b What’s known on the subject? and What does the study add? The results of ICSI using fresh or frozen sperm on the site of sperm retrieval remains controversial with respect to outcome. The results of this study showed no difference in outcome using ICSI either with respect to the site of retrieval or whether the sperm used was fresh or frozen. It also showed that the outcome of ICSI is not related to the underlying cause of the azoospermia.

OBJECTIVES

? To compare the outcome of first‐attempt intracytoplasmic sperm injection (ICSI) ICSI–embryo transfer (ET) cycles using frozen‐thawed testicular sperm (FTTS), fresh testicular sperm (FTS), frozen‐thawed epididymal sperm (FTES) and fresh epididymal sperm (FES) so as to determine which of these has the most successful ICSI outcome with respect to fertilization rate (FR), pregnancy rate (PR) and birth rate. ? To assess the outcomes according to the underlying aetiology of azoospermia.

PATIENTS AND METHODS

? The records of 493 patients undergoing first‐attempt ICSI between 1993 and 2008 were reviewed retrospectively. FTS was used in 112 cycles, FTTS in 43 cycles, FES in 279 cycles, and FTES in 59 cycles. ? Within each group, the aetiology of the azoospermia was recorded according to history, clinical examination and histological analysis (n= 316). ? The FR, clinical PR and delivery rate were calculated for each group with respect to the type of sperm retrieval used.

RESULTS

? Analysis of the data showed no significant differences between any of the four groups in the FR, PR or delivery rate (P > 0.05). ? There were no significant differences seen between fresh sperm (FTS and FES) and frozen sperm (FTTS and FTES) or between epididymal sperm (FES and FTES) and testicular sperm (FTS and FTTS) in any of the outcomes measured (P > 0.05). However, sub‐set analysis showed a statistically higher FR and PR for FTTS over fresh sperm. ? When comparing aetiologies, there was no significant difference in the FR, clinical PR and delivery rate between obstructive azoospermia (OA) and non‐obstructive azoospermia (NOA) groups. However, sub‐set analysis showed a higher PR and birth rate for FTTS over fresh sperm in both OA and NOA groups.

CONCLUSIONS

? The results of the present study suggest that using frozen sperm in ICSI cycles is a reliable and favourable method with the same outcome as fresh sperm. ? Testicular and epididymal sperm have similar ICSI outcomes for both fresh and frozen samples. However, results suggest a tendency for higher PRs and birth rates for frozen than for fresh testicular sperm in both OA and NOA aetiologies. ? The aetiology of azoospermia does not significantly affect the outcome of first‐attempt ICSI. The higher rates in the frozen groups suggest that these patients have had better quality semen when they were initially harvested and frozen.     

睾丸切开显微取精辅助非阻塞性无精子症患者生育

目的:探讨睾丸切开显微取精术在辅助男性非阻塞性无精子症患者生育的效果。方法:采用睾丸切开显微取精术获取精子,结合卵浆内单精子显微注射技术,辅助1例非阻塞性无精子症不育患者人工受精。结果:精子获取成功,结合卵浆内单精子显微注射技术使患者妻子获得妊娠,并成功分娩1健康女婴。结论:睾丸切开显微取精术为非阻塞性无精子症患者生育,提供了一种新的方法。     

经皮睾丸微穿刺活检后冷冻精子的卵胞质内单精子注射

目的:对经皮睾丸微穿刺活检后冷冻保存精子卵胞质内单精子注射(ICSI)治疗非梗阻性无精子症所致不育进行临床总结,并对其影响治疗结果的因素进行探讨。方法:对62例非梗阻性无精子症患者进行经皮睾丸微穿刺活检,发现活动精子者(35例)对睾丸活检组织进行冷冻保存;女方促排卵常规使用促性腺激素释放激动剂(GnRHa)/卵泡刺激素(FSH)/人绒毛膜促性腺激素(hCG)方案,B超监测卵泡发育情况并引导经阴道取卵,冷冻的睾丸组织解冻后行ICSI,良好胚胎进行移植。结果:取卵周期为35个,冷冻的睾丸精子解冻后行ICSI,35个周期进行常规胚胎移植。13例临床妊娠。启动周期、取卵周期与移植周期临床妊娠率均为37.14%(13/35)。结论:经皮睾丸微穿刺活检后ICSI是治疗非梗阻性无精子症所致不育的最主要和有效的方法;活检后对有活动精子的睾丸组织进行冷冻保存不影响治疗结果,可以减少患者睾丸活检的次数,减轻患者痛苦。     

Comparison of the outcome of intracytoplasmic sperm injection in obstructive and non-obstructive azoospermia in the first cycle: a report of case series and meta-analysis

To investigate the outcome of intracytoplasmic sperm injection with fresh and cryopreserved-thawed testicular spermatozoa in the first cycle in patients with obstructive azoospermia (OA) and non-obstructive azoospermia (NOA), a total of 90 cases, 48 OA and 42 NOA were studied. All patients underwent sperm retrieval by testicular sperm extraction (TESE) while their wives received conventional ovarian hyperstimulation. The hormone levels, testicular histology, the rates of sperm retrieval, fertilization, implantation and pregnancy were analysed and evaluated. This study and other four similar studies were subjected to meta-analysis. Sperm retrieval was successful in 100% OA and 61% NOA. Fresh spermatozoa were used in 87.5% and 92.4% of OA and NOA cases respectively; while cryopreserved-thawed spermatozoa were used in 12.5% and 7.6% of OA and NOA, respectively. The fertilization, implantation and clinical pregnancy rates were 65.5%, 15% and 25% respectively in OA group, and 54.2%, 5% and 23.1% respectively in NOA group. Sperm status (fresh or thawed), male partner's age, female age and male serum follicle-stimulating hormone had no significant effect upon fertilization rate, implantation rate, or pregnancy rate per embryo transfer. The results of meta-analysis indicate that there is no statistically significant difference in clinical pregnancy rates between the two groups. There was a significantly higher fertilization rate among OA patients in all analysed studies (95% CI = 14.29-15.71, d.f. 832, T = 1.96). In conclusion, although the fertilization rate was significantly higher in the OA group in our study and from the given meta-analysis, there were some differences as regards pregnancy rates. Although the overall effect was more or less similar pregnancy rates in both subtypes of azoospermia, this may not be true if non-male infertility variables were controlled for in all studies.     

Reliable single sperm cryopreservation in Cell Sleepers for azoospermia management

Conventional sperm freezing methods perform best when freezing sperm samples containing at least hundreds of spermatozoa. In this severe male factor infertility case series, we examined the reproductive outcomes in 12 intracytoplasmic sperm injection cases where spermatozoa used were frozen in Cell Sleepers. Cell Sleepers are novel devices in which individual spermatozoa can be frozen in microdroplets. The case series included five men with obstructive azoospermia, six with nonobstructive azoospermia and one with cryptozoospermia, in whom microscopic sperm retrievals from testicular sperm extraction (TESE), micro‐TESE extracts and a centrifugation procedure resulted in less than 50 spermatozoa. A total of 304 microscopically retrieved spermatozoa were frozen in 20 Cell Sleepers using a rapid manual cryopreservation method. A total of 179 mature oocytes were injected with recovered thawed spermatozoa, resulting in a fertilisation rate of 65.9% (118 of 179), with no total fertilisation failures. In 10 cases, an embryo transfer was performed, three on day 3 and seven on day 5, resulting in a per cycle pregnancy rate of 58.3% (seven of 12). Four of the pregnancies have progressed past 20 gestation weeks. The recovery and use of spermatozoa that were frozen in Cell Sleepers was uncomplicated and effective and eliminated the need to perform any microscopic sperm retrieval procedures on the day of oocyte collection. Modification of the routine sperm cryopreservation methodology to include the use of Cell Sleepers increases the range of sperm samples that can be effectively cryopreserved, to include men with severe male factor fertility.     

Microdissection testicular sperm extraction and IVF-ICSI outcome in nonobstructive azoospermia

We evaluated the efficiency of microdissection testicular sperm extraction (MicroTESE) in patients with nonobstructive azoospermia (NOA) and their pregnancy outcomes in a programme based on in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI). Fifty-six MicroTESE procedures were performed in 53 patients with NOA. Pre-operative levels of luteinising hormone, follicle-stimulating hormone (FSH), testosterone and prolactin were obtained and a Doppler sonography examination was conducted. Sperm retrieval rate, mean age of female partner, mean ICSI and fertilisation rate, number and quality of embryos transferred, implantation, pregnancy and miscarriage rates were calculated. Samples for testicular histological analysis were taken trans-operatively in every case. Sperm retrieval rate, mean ICSI per case and fertilisation rate were 57.1%, 7.4% and 58.4% respectively. A significant difference in pre-operative testicular volume ( P  = 0.001), serum FSH ( P  = 0.008) and total testosterone levels ( P  = 0.021) was found in patients from whom sperm could be retrieved. Mean 1.9 type A embryos were transferred per cycle. Implantation, clinical pregnancy and miscarriage rates were 20%, 40% and 18.7% respectively. It is concluded that MicroTESE is a viable option for men with NOA, offering excellent results in couples undergoing IVF-ICSI. Pre-operative serum FSH, testicular volume and total testosterone levels may have a prognostic value, although more data are needed to determine their significance and whether or not patients should be excluded from an initial sperm retrieval attempt.     

Open testicular mapping: A less invasive multiple biopsy approach for testicular sperm extraction

This study proposes a testicular sperm extraction technique that was inspired by testicular fine-needle aspiration. Here, we have described the technique of open testicular mapping (OTEM) and evaluated the successful sperm recovery in 92 patients with nonobstructive azoospermia (NOA). All patients underwent an OTEM biopsy. Patients were divided into two groups; group I included men with spermatozoa recovered and group 0 included men without spermatozoa recovered. Age, follicle-stimulating hormone (FSH) level and testicular volume were compared between the groups. In 50 of 92 men (54%), viable spermatozoa were found after OTEM. No differences were noted in age, FSH level or testicular volume. Using OTEM, it was possible to retrieve spermatozoa in 54% of the NOA men.