High implantation and pregnancy rates with testicular sperm extraction and intracytoplasmic sperm injection in obstructive and non-obstructive azoospermia

Thirty-two infertile couples with obstructive and non-obstructiveazoospermia were included in this study. Testicular sperm extraction(TESE) was performed in 16 obstructive azoospermic cases wheremicrosurgical sperm aspiration (MESA) or percutaneous spermaspiration (PESA) were impossible because of totally destroyedepididymis and 16 non-obstructive azoospermia cases with severespermatogenetic defect where the testicles were the only sourceof sperm cells. A total of 288 oocytes was obtained from 32females and 84% were injected. The fertilization rates (FR)with 2 pronuclei (PN) and cleavage rate were 50.8 and 68.2%respectively. A total of 15 pregnancies was achieved (53% perembryo transfer), nine from the obstructive and six from thenon-obstructive group. Four pregnancies resulted in clinicalabortion (26.6%). The ongoing pregnancy rate was 39.2% per embryotransfer (ET) and 343% per started cycle. A high implantationrate was also achieved (26.6% in non-obstructive and 30% inobstructive azoospermia group). Using testicular spermatozoain combination with ICSI in both obstructive and non-obstructiveazoospermic groups, high implantation and pregnancy rates canbe achieved.     

Increased sperm motility after in-vitro culture of testicular biopsies from obstructive azoospermic patients results in better post-thaw recovery rate

The objective of this study was to optimize the use of testicular biopsies in 14 patients with obstructive azoospermia. Testicular specimens were retrieved from six patients (group I) and cultured at 32 and 37 degrees C for up to 20 days; changes in percentage motile spermatozoa were compared. In four men of group I, one portion of the specimen was frozen at retrieval, and changes in post-thaw motility after 24 h of culture at 37 degrees C were recorded. In the other eight patients (group II), testicular specimens were frozen at retrieval and after 72 h culture at 37 degrees C. Pre and post-freezing motility and post-thaw recovery rate were compared. No significant differences were observed until day 8 in the improvement of motility between 32 and 37 degrees C in-vitro culture. Maximum motility was reached, under both conditions, between 48 h and 72 h. Post-thaw 24 h culture at 37 degrees C of specimens frozen at retrieval did not improve motility; however, 72 h pre-freezing culture significantly improved initial motility (P: < 0.01), post-thaw motility (P: < 0.01) and post-thaw recovery rate (P: < 0. 001). The higher recovery rate of samples frozen 3 days after retrieval allows more economical use of the tissue that is available.     

Fertility with testicular sperm extraction and intracytoplasmic sperm injection in non-obstructive azoospermic men

In non-obstructive azoospermia spermatozoa can usually onlybe isolated from the testicles, and thus the most promisingtreatment model is testicular sperm extraction (TESE). Hormoneconcentrations, testicular volume determinations and testicularbiopsy results are not uniform enough to select potential candidatesfor successful TESE and intracytoplasmic sperm injection (ICSI)approaches in advance. The aim of this study was to assess theefficacy of using ICSI with testicular spermatozoa in casesof non-obstructive azoospermia and to compare the inclusioncriteria and sperm existence in the testicles in sperm obtainableand non-obtainable groups. All men showed either complete orincomplete (n = 14) maturation arrest in spermatogenesis, severehypospermatogenesis (n = 10) or Sertoli cell-only syndrome (n= 5) in their testicular biopsies. Only 14 out of a total of29 men provided enough spermatozoa for the ICSI procedure, whileno spermatozoa were found in the testicular samples of the remaining15 men. Out of 123 oocytes obtained from 14 females, 101 wereinjected with the husbands' testicular sperm cells. Total fertilizationfailure was observed in three cases. Of 39 oocytes fertilized,38 cleaved. The fertilization and cleavage rates were 38.6 and97.4% respectively. The pregnancy rate was 20.7% per initiatedcycle. In the group from whom spermatozoa were obtainable, thepregnancy rate was 42.9% per initiated cycle and 54.5% per embryotransfer. A total of six pregnancies were achieved, of whichtwo Were twins and four were singletons. One singleton pregnancyresulted in abortion in the first trimester. There was no statisticaldifference concerning the serum follicle stimulating hormoneconcentration, testicular volume and biopsy results in groupsin which spermatozoa were obtainable or not. In conclusion,although the association of TESE with ICSI obtained pregnanciesfor some patients with non-obstructive azoospermia, furtherstudies are needed to determine the inclusion criteria for successfulTESE.     

Outcome of intracytoplasmic sperm injection with testicular spermatozoa in obstructive and non-obstructive azoospermia

From 1 August 1993 until 30 September 1994, 69 couples sufferingfrom azoospermia underwent testicular sperm extraction and intracytoplasmicsperm injection. In 50 couples with obstructive azoospermiaa total of 631 meta-phase-II oocytes were injected after testicularsperm extraction yielding a 2-PN fertilization rate of 57%.In female patients <40 years of age an ongoing pregnancyrate per transfer of 42% (14/33) was obtained. So far, eighthealthy babies have been born, including two singletons andthree twin gestations. In 19 couples with non-obstructive azoospermiaa total of 264 metaphase-II oocytes were injected after testicularsperm extraction, yielding a 2-PN fertilization rate of 58%.An ongoing pregnancy rate per transfer of 31% (5/16) was established.So far, six healthy babies have been born including one singleton,one twin and one triplet gestation.     

Correlation between testicular histology and outcome after intracytoplasmic sperm injection using testicular spermatozoa

A comprehensive study is presented of a series of 124 infertilemen undergoing testicular sperm retrieval for intracytoplasmicsperm injection (ICSI). In this study we correlated the histologicalchanges observed in the testicular tissue with the results ofthe wet preparation and the outcome after ICSI using testicularspermatozoa. In all patients with normal spermatogenesis andhypospermatogenesis spermatozoa were recovered from the wetpreparation. The sperm recovery rate was 84% in patients withincomplete germ-cell aplasia and maturation arrest, while inpatients with complete germ-cell aplasia or maturation arrestthis figure was 76%. In these patients more specimens were sampledand fewer spermatozoa were recovered. Since no spermatozoa wererecovered in only 10 patients, ICSI with testicular sperm wasperformed in the remaining 114 couples (91.9%). The normal fertilizationrate was 57.8%. The fertilization rate was significantly lowerin couples among whom the husband showed germ-cell aplasia andmaturation arrest. Overall, 55.2% of normally fertilized oocytesdeveloped into embryos showing 50% of anucleate fragments. Therewere no major differences between the different histologicalcategories in terms of embryonic development in vitro. The overallpregnancy rates per testicular sperm extraction (TESE) procedure,per ICSI procedure and per transfer were respectively 36.3,39.5 and 43.7%. The overall implantation rate per embryo (sacs/embryosreplaced) was 20.3%. A lower implantation rate was observedin couples among whom the husband had maturation arrest (notstatistically significant). The above data show that testicularbiopsies may have an important therapeutic role in the managementof infertility in azoospermic patients.     

Pregnancies after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa

In 25 patients (14 suffering from obstructive azoospermia, sixfrom non-obstructive azoospermia, three from astheno-azoospermiaand two from absence of ejaculation) spermatozoa were extractedfrom testicular biopsies. Intracytoplasmic sperm injection (ICSI)with fresh testicular spermatozoa was performed in 18 cases;spermatozoa in excess were cryopreserved in pills. No pregnancieswere achieved. In the remaining seven patients, testicular spermatozoawere retrieved and cryopreserved during a diagnostic testicularbiopsy. After thawing, sperm motility was assessed in 17 cases(68%), and 18 ICSI with cryopreserved testicular spermatozoawere performed. The mean two-pronuclear (2PN) fertilizationrate was 59%, the mean cleavage rate was 92%, and six clinicalpregnancies were achieved, all of them still ongoing (pregnancyrate 33%). A comparison of the results of ICSI carried out withfresh or cryopreserved testicular spermatozoa showed that themean 2PN fertilization rates per cycle (53 compared with 55%),mean cleavage rates per cycle (99 compared with 96%) and embryoquality were not significantly different In conclusion, cryopreservationof testicular spermatozoa is feasible, even in patients withnon-obstructive azoospermia, and the results of ICSI with frozen-thawedtesticular spermatozoa are similar to those obtained using freshtesticular spermatozoa. Cryopreservation of testicular spermatozoamay avoid repetition of testicular biopsies to retrieve spermatozoafor successive ICSI cycles in patients in whom the only sourceof motile spermatozoa is the testicle.     

Fertilization and pregnancy outcome with intracytoplasmic sperm injection for azoospermic men

The evident ability of the intracytoplasmic sperm injection (ICSI) procedure to achieve high fertilization and pregnancy rates regardless of semen characteristics has induced its application with spermatozoa surgically retrieved from azoospermic men. Here, ICSI outcome was analysed in 308 cases according to the cause of azoospermia; four additional cycles were with cases of necrozoospermia. All couples were genetically counselled and appropriately screened. Spermatozoa were retrieved by microsurgical epididymal aspiration or from testicular biopsies. Epididymal obstructions were considered congenital (n = 138) or acquired (n = 103), based on the aetiology. Testicular sperm cases were assessed according to the presence (n = 14) or absence (n = 53) of reproductive tract obstruction. The fertilization rate using fresh or cryopreserved epididymal spermatozoa was 72.4% of 911 eggs for acquired obstructions, and 73.1% of 1524 eggs for congenital cases; with clinical pregnancy rates of 48.5% (50/103) and 61.6% (85/138) respectively. Spermatozoa from testicular biopsies fertilized 57.0% of 533 eggs in non-obstructive cases compared to 80.5% of 118 eggs (P = 0.0001) in obstructive azoospermia. The clinical pregnancy rate was 49.1% (26/53) for non-obstructive cases and 57.1% (8/14) for testicular spermatozoa obtained in obstructive azoospermia, including three established with frozen-thawed testicular spermatozoa. In cases of obstructive azoospermia, fertilization and pregnancy rates with epididymal spermatozoa were higher than those achieved using spermatozoa obtained from the testes of men with non-obstructive azoospermia.     

Conventional multiple or microdissection testicular sperm extraction: a comparative study

BACKGROUND: Testicular sperm extraction (TESE) with ICSI is becoming the first-line treatment for non-obstructive azoospermia (NOA). Recently, the sperm retrieval rate (SRR) by microdissection TESE was reported to be higher than by conventional TESE. However, a comprehensive comparison between multiple and microdissection TESE patients including histological findings has not been reported. METHODS: Patients with NOA who underwent microdissection TESE (n = 56) or multiple TESE (n = 37) were compared. Pre-operative characteristics were similar between groups. In addition, microscopic findings during microdissection TESE also were investigated. RESULTS: Operative time was significantly longer for microdissection TESE than for multiple TESE. Histological examination suggested that spermatogenesis was relatively more impaired in the microdissection TESE group than in the multiple TESE group. Despite this, SRR by microdissection TESE (42.9%) appeared higher than by conventional TESE (35.1%) although this observation failed to reach statistical significance. Seventeen of 26 patients (65.4%) with heterogeneous tubule were successful for sperm retrieval. No severe operative complications occurred in any patient in either group, and no patient required post-operative hormone replacement to treat hypogonadism. CONCLUSIONS: Microsurgical technique is safe and may improve SRR for TESE in a variety of patients with NOA, especially patients with heterogeneous testicular tubules.     

Aggressive sperm immobilization prior to intracytoplasmic sperm injection with immature spermatozoa improves fertilization and pregnancy rates

This study was conducted to determine whether the mode of spermimmobilization prior to intracytoplasmic sperm injection (ICSI)influences fertilization by immature spermatozoa. Of the 837ICSI cycles evaluated, 81 were performed with epididymal ortesticular spermatozoa; 35 cycles with epididymal spermatozoaimmobilized in the standard fashion resulted in fertilizationand pregnancy rates of 48.3 and 51.4% respectively. When a moreaggressive sperm immobilization technique (i.e. permanentlycrimping the sperm fiagellum between the midpiece and the restof the tail) was applied in 17 cycles, the resultant fertilizationand pregnancy rates were significantly (P < 0.05) higher:82.0 and 82.4% respectively. Similar increases in fertilizationand ensuing pregnancy rates were also observed in ICSI cycleswith the aggressive immobilization of frozen-thawed epididymalspermatozoa (eight cycles) versus standard immobilization (16cycles). However, the fertilization rates for ICSI using testicularspermatozoa (five cycles) were basically the same, regardlessof the immobilization technique. Furthermore, for ejaculatedspermatozoa (756 cycles), the fertilization rates followingaggressive sperm immobilization were also positively affected(73.4%), although no statistical differences in the clinicalpregnancy rates were found. Because aggressive immobilizationappears to affect sperm membrane pennea-bilization, the enhancedfertilization patterns observed in immature spermatozoa followingaggressive immobilization may suggest a different membrane constitutionin these spermatozoa. These findings indicate that immaturegametes may require additional manipulation to enhance the post-ICSIevents essential for adequate nuclear decon-densation.     

Andrology: CASE REPORT Pregnancy after intracytoplasmic sperm injection of spermatozoa extracted from frozen-thawed testicular biopsy

The case report illustrates the successful application of anew method of sperm extraction from a frozen-thawed testicularbiopsy specimen within an established programme of intracytoplasmicsperm injection.     

High fertilization and implantation rates after intracytoplasmic sperm injection

Previously reported better fertilization rate after intra-cytoplasmicsingle sperm injection (ICSI) than after subzonal inseminationof several spermatozoa was confirmed in a controlled comparisonof the two procedures in 11 patients. Intracytoplasmic sperminjection was carried out in 150 consecutive treatment cyclesof 150 infertile couples, who had failed to have fertilizedoocytes after standard in-vitro fertilization (IVF) proceduresor who were not accepted for IVF because not enough motile spermatozoawere present in the ejaculate. A single spermatozoon was injectedinto the ooplasm of 1409 metaphase II oocytes. Only 117 oocytes(8.3%) were damaged by the procedure and 830 oocytes (64.2%of the successfully injected oocytes) had two distinct pronucleithe morning after the injection procedure. The fertilizationrate was not influenced by semen characteristics. After 24 hof further in-vitro culture, 71.2% of these oocytes developedinto embryos, which were transferred or cryopreserved. Only15 patients did not have embryos replaced. Three-quarters ofthe transfers were triple-embryo transfers. High pregnancy rateswere noticed since 67 pregnancies were achieved, of which 53were clinical, i.e. a total and clinical pregnancy rate of 44.7%and 35.3% per started cycle and 49.6% and 39.2% per embryo transfer.A total of 237 supernumerary embryos were cryopreserved in 71treatment cycles.     

显微取精术在非梗阻性无精子症患者中的临床应用及结局

目的观察睾丸显微取精术在非梗阻性无精子症患者中的临床应用及其结局。方法17例患者在配偶同步促排卵取卵前一天行睾丸显微切开取精术,获得精子患者行卵胞浆内单精子显微注射(ICSI)助孕,分析其受精率、可用胚胎率和临床妊娠结局。结果17例患者显微取精,13例成功获取精子,精子获得率为76.5%。这13对夫妇ICSI受精率为74.8%,可用胚胎率49.5%。3例新鲜周期移植,全部成功妊娠,1例已分娩1健康婴儿,另外2例持续妊娠中。其余10例行全胚冷冻,4例分别于胚胎冻存3个月后行解冻复苏移植,3例成功妊娠,2例已分娩(其中一例为非嵌合型Klinefelter综合征患者),另外1例持续妊娠中。结论睾丸显微切开取精术是使NOA患者和非嵌合型Klinefelter综合征患者成功获得自己遗传学子代的有效方法,有较高的精子获得率,联合ICSI技术、全胚冷冻-复苏移植可以获得较高的临床妊娠率。     

Testicular sperm extraction: impact of testicular histology on outcome, number of biopsies to be performed and optimal time for repetition

Testicular sperm extraction (TESE) may not always be successful in patients with non-obstructive azoospermia, as they only have minute foci of active spermatogenesis from which a tiny number of spermatozoa can be extracted. The aim of this study was to find the percentile incidence of successful TESE in non-obstructive azoospermia patients in relation to various histopathological patterns and the number of performed biopsies, and to determine the optimal time needed for repetition. A total of 216 patients underwent bilateral testicular biopsy taking a single piece from each testis for sperm retrieval and pathological evaluation. In another 100 patients, the same procedure was done but taking multiple samples (maximum four samples/testis). Spermatozoa were successfully retrieved from 37.5 and 49% of patients who supplied single and multiple samples respectively. TESE was significantly higher when multiple samples were taken in all histopathological groups except for Sertoli cell-only syndrome, tubular sclerosis and Klinefelter's pattern. Twenty-seven patients underwent repeated TESE for ICSI between 1 and 24 months from the first procedure; all of them had easy sperm retrieval during the first procedure. Although sperm retrieval was successful in 75 and 94.7% of patients who underwent the second attempt, before and after 3 months respectively, a second TESE was usually more difficult and necessitated multiple sampling.     

Case report: Pregnancy resulting from intracytoplasmic injection of spermatozoa from a frozen-thawed testicular biopsy specimen

A testicular biopsy specimen was taken in connection with scrotalexploration of a healthy 35 year old man who had azoospermia.Bilateral severe scarring of unknown aetiology was found inthe exploration, and no epididymal spermatozoa could be obtained.Spermatozoa from the fresh biopsy specimen were used for intracytoplasmicsperm injection (ICSI) on the same day. Two-embryo transferresulted in biochemical pregnancy. The rest of the biopsy specimenwas frozen as small pieces of tissue using glycerol as a cryoprotectant.ICSI was then performed with spermatozoa prepared from the frozen-thawedtissue. One embryo was obtained and transferred. The transferresulted in pregnancy, and a living fetus was seen in ultrasoundscans at the seventh and 16th weeks of pregnancy. It is possibleto avoid repeated testicular biopsies by using cryopreservationof testicular tissue.     

Blastocyst transfer following intracytoplasmic injection of ejaculated, epididymal or testicular spermatozoa

Recent studies indicate a strong paternal influence on embryo development and progression of the embryo to the blastocyst stage. The aim of this study was to compare, during extended culture, the in-vitro development of embryos resulting from intracytoplasmic sperm injection (ICSI) of ejaculated spermatozoa (group 1, n = 347), epididymal (group 2, n = 22) or testicular (group 3, n = 18) spermatozoa from obstructive azoospermic and testicular spermatozoa from non-obstructive azoospermic (group 4, n = 31) subjects. Fertilization and blastocyst formation rates were significantly lower in group 4 (P < 0.05). The incidence of expanded and hatching blastocysts was significantly lower in group 4 (P < 0.05). Overall in 93.2% ejaculate ICSI cycles, blastocysts were transferred on day 5. This was significantly higher than the 62% day 5 transfers in the non-obstructive azoospermic group (P < 0.05). Implantation rate per embryo was significantly higher in the ejaculate ICSI group compared with the other groups (P < 0.05). Clinical pregnancy per transfer was similar between groups; however, significantly fewer multiple pregnancies were encountered in the non-obstructive azoospermic group (P < 0.01). In conclusion, the source of the spermatozoa, most likely to be indicative of the severity of spermatogenic disorder, affects the rate of blastocyst formation and blastocyst implantation. Spermatozoa from non-obstructive azoospermic subjects, when utilized for ICSI, result in embryos that progress to the blastocyst stage at a lower and slower rate and implant less efficiently.     

Diagnostic epididymal and testicular sperm recovery and genetic aspects in azoospermic men.

Various procedures for sperm recovery in azoospermic men have been described, from open testicular biopsy to simple needle aspiration from the epididymis and the testis. Fifty-one obstructive and 86 non-obstructive azoospermic men were treated to compare the recovery of spermatozoa obtained by percutaneous aspiration from the epididymis (PESA) and aspiration/extraction from the testis (TESA, TESE) with histopathology. If TESA failed, the work up proceeded with TESE. All patients were karyotyped. Spermatozoa were recovered by PESA or TESA in all obstructive men (51/51 patients). In 22 out of 86 patients with non-obstructive azoospermia, testicular spermatozoa could be successfully recovered by TESA. In five additional patients TESE was successful in recovering spermatozoa where TESA had failed. In 43 patients, neither TESA nor TESE was successful. Sixteen patients chose not to proceed with TESE. Seven out of 86 patients had an abnormal karyotype in the non-obstructive group (8%), none in the obstructive group. In the non-obstructive patient group testicular histopathology showed hypospermatogenesis, incomplete maturation arrest and germ cell aplasia with focal spermatogenesis in cases where spermatozoa were recovered and complete germ cell aplasia, complete maturation arrest and fibrosis in cases where no spermatozoa were found. Spermatozoa were recovered by PESA or TESA from all patients with obstructive azoospermia and from approximately 40% of patients with non-obstructive azoospermia by TESA or TESE. Retrieval of viable spermatozoa in the infertility work-up was highly predictable for sperm recovery in subsequent ICSI cycles. TESA performed under local anaesthesia seems almost as effective as more invasive procedures in recovering testicular spermatozoa, both in obstructive and non-obstructive azoospermic men.     

Pregnancies achieved after frozen-thawed pronuclear oocytes obtained by intracytoplasmic sperm injection with spermatozoa extracted from frozen-thawed testicular tissues from non-obstructive azoospermic men.

The use of frozen-thawed testicular tissue as a source of spermatozoa for intracytoplasmic sperm injection (ICSI) in non-obstructive azoospermia yields favourable fertilization and pregnancy rates while avoiding both repetitive biopsies and unexpected cycle cancellations. Spermatozoa were obtained from frozen-thawed testicular biopsy specimens from 67 non-obstructive azoospermic men. Following fertilization, supernumerary two pronuclear (2PN) oocytes were frozen. After thawing, 17 cycles of embryo transfer were carried out with a mean number of 2.7 embryos and a mean cumulative embryo score (CES) of 18.3 per transfer. The clinical pregnancy and implantation rates per transfer in these cycles (23.5 and 8.3% respectively) were comparable to those of fresh embryo transfers (35.7 and 12.7% respectively) with a mean number of 2.7 embryos and a mean CES of 28.7 per transfer. Abortion rates, although higher with cryopreserved 2PN oocytes were not significantly different. With this approach, cryopreservation of supernumerary 2PN oocytes can be used to improve the cumulative pregnancy rates in a severely defective spermatogenetic population. To our knowledge, these are the first pregnancies reported which have been obtained by the transfer of cryopreserved pronuclear oocytes obtained from ICSI using cryopreserved testicular spermatozoa.     

Enzymatic digestion of testicular tissue may rescue the intracytoplasmic sperm injection cycle in some patients with non-obstructive azoospermia

Recovery of testicular spermatozoa from azoospermic patientswith testicular failure, followed by intracytoplasmic sperminjection (ICSI) is a recent advance in the treatment of maleinfertility. In most cases, free spermatozoa are recovered fromtesticular tissue after mechanical mincing of multiple biopsies.Testicular sperm retrieval, however, remains unsuccessful in30–50% of male patients suffering from Sertoli cell-onlysyndrome and maturation arrest. In this study, a strategy wasdeveloped in order to maximize the chance of sperm retrievalin difficult cases of testicular failure. The ultimate stepwas the use of enzymatic procedures (collagenase type IV) todissociate the testicular tissue completely. Testicular tissueof 41 patients for whom no spermatozoa were found after mechanicalmincing of the testicular tissue was investigated. In 14 outof the 41 cases (34%), enough spermatozoa for ICSI were foundafter fine mincing of multiple biopsies and several hours' searchin the cell suspension treated with the erythrocyte-lysing buffer(ELB). In 27 out of the 41 patients, no spermatozoa were foundeven after the use of ELB. In seven out of these 27 failures(26%), spermatozoa for ICSI were retrieved after enzymatic dissociationof the residual minced tissue pieces, thus making ICSI possibledespite failure to find spermatozoa with conventional mincing.From this study, we may conclude that enzymatic digestion oftesticular tissue is easy to perform, is not time-consumingand constitutes a successful method in reducing the sperm recoveryfailures in patients with non-obstructive azoospermia.     

Case report: Changes in motility patterns during in-vitro culture of fresh and frozen/thawed testicular and epididymal spermatozoa: implications for planning treatment by intracytoplasmic sperm injection

The present report describes the motility changes in vitro (percentagemotile and progressively motile) of freshly collected testicularand epididymal spermatozoa and following freeze/thaw of thesame spermatozoa from a man with obstructive azoospermia. Washedspermatozoa were cultured in micro droplets under paraffin oilor in test tubes using HEPES-buffered or bicarbonate-bufferedmedium containing 10% human serum. In fresh testicular spermcultures 60–65% of the sperm cells became motile within2 days of culture; the motility was maintained for a further4–5 days before a decline was observed. The progressivemotility unproved markedly on the third day of culture and itpeaked around day 5. Only a small number of frozen/ thawed testicularspermatozoa became motile during in-vitro culture (15–20%)and the motility was maintained for only 2–3 days beforeit declined. Furthermore, only 10–12% of the spermatozoashowed progressive motility. Spermatozoa recovered from micro-epididymalsperm aspiration (MESA) showed a gradual decrease in progressivemotility and in 5 days all sperm cells were found to be immotilein both freshly collected and frozen/thawed spermatozoa. Allculture systems supported sperm motility. It is clear that testicularspermatozoa, particularly from men with obstructive azoospermia,can be collected and maintained in vitro for up to 1 week beforethe oocyte retrieval but when frozen testicular or epididymalspermatozoa are used it is more reliable to thaw these spermatozoaon the day of intracytoplasmic sperm injection.     

Outcome of testicular sperm retrieval procedures in non-obstructive azoospermia: percutaneous aspiration versus open biopsy

The aim of this study was to evaluate whether the extraction of testicular spermatozoa with percutaneous versus open biopsy has an effect on the treatment outcome with intracytoplasmic sperm injection (ICSI) in men with non-obstructive azoospermia. Regardless of testicular size, follicle stimulating hormone concentration, and previous biopsy result, percutaneous testicular sperm aspiration (PTSA) using a 21-gauge butterfly needle was attempted first and if this failed testicular sperm extraction (TESE) was performed. In 63 men spermatozoa were found with PTSA whereas in 228 men TESE had to be undertaken. More men in the PTSA group had previously been diagnosed with hypospermatogenesis (82 versus 50%). Compared with the PTSA group, more men in the TESE group had germ cell aplasia (27 versus 10%) or maturation arrest (22 versus 8%). There was no difference between the groups regarding mean age of men and their partners, duration of stimulation, oestradiol concentration on the day of human chorionic gonadotrophin, number of oocytes retrieved, fertilization rate, and embryo quality between the two groups. The number of embryos transferred (4.38 versus 3.90) was significantly higher in the PTSA group (P < 0.05), reflecting the increased number of embryos available for transfer. Implantation rate per embryo was 20.7% in the PTSA and 13.3% in the TESE group (P < 0.05). Clinical pregnancy rates were 46 and 29% in the PTSA and TESE groups respectively (P < 0.05). Clinical abortion rates were similar (21.2 versus 24%). It is concluded that in men with non-obstructive azoospermia, easier sperm retrieval, which is most likely indicative of a more favourable histopathology, is associated with higher implantation rates per embryo.