Microepididymal Sperm Aspiration Research Articles

#42 : Comparison of Fertilization, Blastocyst Utilization, Clinical Pregnancy Rate (cPR), Implantation Rate (IR) and On-Going Pregnancy Rate Using Fresh and Frozen-Thawed Micro-Epididymal Sperm Aspiration (MESA) Sperm

Background and Aim: To compare the fertilization, blastocyst utilization, clinical pregnancy rate (cPR), implantation rate (IR) and on-going clinical pregnancy rate between fresh versus frozen-thawed micro-epididymal aspirated sperm (MESA) in intra-cytoplasmic sperm injection (ICSI) cycle. Methods: A retrospective study of a total of 27 MESA-ICSI cycles done from the year 2019 to 2022 was carried out. Patients were categorized into fresh MESA-ICSI group (n=16) and frozen-thawed MESA-ICSI group (n=11). The maternal mean age of both groups was 33.0 and 32.5 respectively. For fresh MESA-ICSI group, retrieved sperm were being injected on the same day while frozen-thawed MESA-ICSI group, sperm sample were being frozen after the MESA procedure and utilized in the future. Sample was processed using sperm medium (COOK MEDICALⓇ). All embryos were cultured in time-lapse monitoring incubator and assessed according to modified Gardner’s grading system. On day 5/6, blastocyst range from grade A to C will be vitrified for frozen embryo transfer (FET). A total of 22 FET procedures were done, and statistical analysis was performed using Fisher’s exact test, p<0.05. The mean number of embryos transferred for MESA-ICSI group and frozen-thawed MESA-ICSI group are 1.43 and 1.13 respectively. Results: There is no significant difference in the fertilization rate between fresh MESA-ICSI, 71.1% (106/149) and frozen-thawed MESA-ICSI group, 65.4% (70/107). The blastocyst utilization rate between fresh and frozen-thawed MESA-ICSI group are also similar, 45.3% (48/106) and 51.4% (36/70) respectively. Besides, the cPR shown higher in the fresh MESA-ICSI group, 64.3% (9/14) compared to frozen-thawed MESA-ICSI group, 37.5% (3/8). The IR, 50.0% (10/20) and on-going pregnancy rate, 57.1% (8/14) were higher in the fresh MESA-ICSI group compared to frozen-thawed MESA-ICSI group, 33.3% (3/9) and 12.5% (1/8) respectively, p<0.05. Conclusion: The results show the trend of higher cPR, IR and on-going pregnancy rate in fresh MESA-ICSI group compared to frozen-thawed MESA-ICSI group. However, further study is required to conduct in a larger population in order to obtain a significant result.

(284) A Systematic Review of Loco-sedative Anesthesia for Penile and Scrotal Surgery

Abstract Introduction The current standard of care for patients undergoing the majority of scrotal and penile procedures is to administer general or spinal anesthesia. The use of general or spinal anesthesia can lead to several complications, long wait times and is not feasible in many rural and remote areas. Objective The objective of this study is to perform a systematic review of the characteristics and outcomes of conscious sedation and local anesthesia for penile and scrotal urologic procedures. Methods A systematic search was conducted on PubMed, and Scopus databases following PRISMA criteria from inception to August 2021. Articles were included if they were prospective, randomized or non-randomized controlled trials that used local anesthetic or conscious sedation for penile or scrotal interventions. Additionally, included studies provided primary data on the use loco-sedative anesthesia and the efficacy and complications from the procedures. Results Nine articles with 4511 patients were included in the study. Procedures examined included vasectomies, adult circumcisions, varicocelectomies, urethrocutaneous fistula repair, microepididymal sperm aspiration and testicular sperm extraction. Of the studies that reported age, the mean patient age was 29.3 years old. The most common anesthetic and analgesic relief was the use of local anesthetic with 1% lidocaine. Only one study used conscious sedation whereas the rest used local anesthetics. However, there was significant heterogeneity in the type of local or conscious sedation method and whether there was a combination was used. 44.4% of the studies used the visual analog scale (VAS) for their primary endpoint. All the studies reported 100% successful procedures with no sedative complications. Conclusions Given the high efficacy rates, loco-sedative anesthesia is a promising technique for penile and scrotal urologic interventions. However, the current body of literature has limited data on a wider array of procedures. Further controlled studies examining the use of loco-sedative anesthesia for a larger variety of penile and scrotal interventions are recommended. This review also demonstrates the significant heterogeneity on how sedation may be assessed and the variety of local anesthetics that can be used to substitute for general or spinal anesthesia. Disclosure No

Sperm chromosome abnormalities in patients with normal karyotype and in translocation carriers: clinical relevance for assisted reproductive technology

Research questionWhat is the proportion of chromosomally abnormal spermatozoa in men with a history of reproductive failure, including patients with normal karyotype and carriers of translocations? Should this analysis be included in a clinical setting to define the best treatment options for infertile couples? DesignAneuploidy for chromosomes XY, 13, 15, 16, 17, 18, 21, 22 was tested by fluorescent in-situ hybridization (FISH) in 1665 samples from couples with normal karyotype having had at least three previous IVF failures, miscarriages, or both (group-A). A FISH test was also carried out in 76 samples from carriers of translocations (group B) to detect the proportion of spermatozoa with unbalanced rearrangement. ResultsIn group A, the lowest incidence of aneuploid sperm cells was found in men with normozoospermia (1.3%, range 0.09–6.31%) compared with men with moderate oligoasthenoteratozoospermia (2.1%, range 0.41–16.6%, P < 0.001), severe oligoasthenoteratozoospermia (4.7%, range 0.53–30.77, P < 0.001), microepididymal sperm aspiration (3.1%, range 1.19–24.24, P < 0.001) and testicular sperm extraction samples (5.8%, range 1.54–33.3, P < 0.001). In group B, the proportion of spermatozoa with unbalanced rearrangement was significantly higher in reciprocal (63%, range 10.0–87.6%) than in Robertsonian translocations (16%, range 4.3–51.0%, P < 0.001). ConclusionsPatients with poor prognosis of term pregnancy tend to generate high proportions of chromosomally abnormal spermatozoa, especially in severe male factor cases. Corresponding frequencies occur at wide ranges; therefore, the FISH test is needed to assess the proportion of spermatozoa with altered chromosome condition. A flowchart, which included the FISH test, was designed to assist clinicians guide couples with poor prognosis of pregnancy, on the most indicated treatment options.

Office-based andrology and male infertility procedures-a cost-effective alternative.

BackgroundFrom 2014–2016, our clinical practice progressively incorporated several male infertility and andrology procedures performed under local anesthesia, including circumcision, hydrocelectomy, malleable penile prostheses, orchiectomy, penile plication, spermatocelectomy, testicular prostheses, varicocelectomy, vasectomy reversal (VR), and testicular and microepididymal sperm aspiration (TESE/MESA). Given the observed outcomes and potential financial and logistical benefits of this approach for surgeons and patients, we sought to describe our initial experience.MethodsA retrospective analysis was performed of all andrologic office-based (local anesthesia only) and select OR (general or monitored anesthesia care) procedures performed from 2014–2016. Financial and outcomes analyses were performed for infertility cases due to the homogeneity of payment modalities and number of cases available. Demographic, clinicopathologic, and procedural costs (direct and indirect) were reviewed and compared.ResultsA total of 32 VRs, 24 hydrocelectomies, 24 TESEs, 10 circumcisions, 9 MESA/TESEs, 4 spermatocelectomies, 3 orchiectomies (1 inguinal), 2 microTESEs, 2 testicular prostheses, 1 malleable penile prosthesis, 1 penile plication, and 1 varicocelectomy. Compared to the OR, male infertility procedures performed in the clinic with local anesthesia were performed for a fraction of the cost: MESA/TESE (78% reduction), TESE (89% reduction), and VR (62% reduction). All office-based procedures were completed successfully without significant modifications to technique. Outcomes were similar between the office and OR including operative time (VR: 181 vs. 190 min, P=0.34), rate of vasoepididymostomy (VE) (23% vs. 32%, P=0.56), total sperm counts (72.2 vs. 50.9 million, P=0.56), and successful sperm retrieval (MESA/TESE 100% vs. 100%, P=1.00; TESE 80% vs. 100%, P=0.36). To our knowledge, the current study also represents the first report of office-based VE under local anesthesia alone. For hydrocelectomy procedures, recurrence (4%) and hematoma (4%) rates were low (mean 4.2 months follow-up), although this likely relates to modifications with technique and not the anesthesia or operative setting. Overall, when given the choice, 86% of patients chose an office-based approach over the OR.ConclusionsOffice-based andrology procedures using local anesthesia may be successfully performed without compromising surgical technique or outcomes. This approach significantly reduces costs for patients and the overall healthcare system and has become our treatment modality of choice.

Azoospermic HIV-1 infected patients wishing to have children: proposed strategy to reduce HIV-1 transmission risk during sperm retrieval and intracytoplasmic sperm injection: Case Report

To date, assisted reproductive technology (ART) with sperm washing is offered to serodiscordant couples with an human immunodeficiency virus-1 (HIV-1) infected male partner in order to have a child while reducing the risk of transmission to the woman. However, ART programmes are not possible if the man is azoospermic. We report here the first birth following intracytoplasmic sperm injection (ICSI) using frozen epididymal spermatozoa obtained after surgical sperm retrieval in a HIV-1 infected man with obstructive azoospermia. METHODS; Sperm obtained by micro-epididymal sperm aspiration was frozen after density gradient preparation and tested for HIV-RNA and DNA. ICSI with frozen sperm was performed. A twin pregnancy was obtained following ICSI. Two healthy girls were born. Maternal HIV-1 RNA and HIV-1 serology were negative during pregnancy and at delivery. This case report demonstrates that ART is possible in azoospermic HIV-1 infected men. On the basis of current knowledge, we propose a strategy to reduce HIV-1 transmission risk during sperm retrieval and ICSI in couples where the man is HIV-1 infected and azoospermic.

New Approach of Intracytoplasmic Sperm Injection With Epididymal Sperm by Retrograde Epididymal Sperm Aspiration for Multiple Failures in ART

Male factor infertility has undergone significant changes in the last few years. The treatment for severe oligozoospermia, and obstructive azoospermia, however, are still difficult. The introduction of intracytoplasmic sperm injection (ICSI) has resulted in better fertilization and pregnancy rates in cases with severe male factor infertility. Several techniques exist to obtain the sperm of severe oligozoospermia and obstructive azoospermia. However the use of a micro-operation scope and fine technique are needed for micro-epididymal sperm aspiration (MESA) and testicular sperm extraction (TESE). There are some risks to be expected such as subcutaneous bleeding in the percutaneous sperm aspiration (PESA). In order to obtain the sperm of severe male factor, the procedure of retrograde sperm aspiration (RESA) was designed and efficacy of RESA was evaluated in this study. RESA was undergone in severe male factor infertility couples who failed pregnant in several ICSI attempts with ejaculated spermatozoa. First, the upper side of the scrotum is opened 1cm and vas deferens is lifted. The G-24 elastic catheter is inserted into the vas toward the epididymis. Then, 0.3∼0.5ml of saline is injected and then aspirated the saline. The small number of epididymal sperms is collected with the saline, twenty -seven patients with severe male factor underwent RESA and ICSI was performed (RESA-ICSI group). Fifty-four patients (as control group), who failed pregnant at least .ve times by ICSI, underwent the ICSI with ejaculated sperm. The mean female age of RESA-ICSI group and control group were 35.1±3.3 years and 35.1±3.4 respectively. The previous cycle of ART in RESA-ICSI group was 10.5±7.5 and in control group was 10.0±6.7. The pregnancy rate per embryo transfer was 18.8% in RESA-ICSI group and 15.7% in control group. The pregnancy rate per cases of RESA-ICSI group (36.0%) tended to be higher than the control group (25.5%, not significantly). To pursue the sperm factors which reflect the efficacy of RESA, Kruger’s strict criteria (s.c.), motility rate, anomaly rate were analyzed. In the group of s.c. below 6%, pregnancy rate showed higher tendency in RESA-ICSI group as compared control group (42.1% vs. 18.2%, respective). In the motility rate patient below 50%, RESA-ICSI group showed the higher pregnancy rate tendency (43.8%) than control group (18.5%). In anomaly rate, patient below 70% showed significant higher pregnancy rate in RESA-ICSI group as compared control group (66.7% vs. 12.5%, P<0.05). RESA is a simple, safe and effective method to obtain the sperm of severe male factor infertility. It was suggested that ICSI with epididymal sperm was an effective treatment for multiple failures in some male factor patients.

303 INDUCED MOTILITY OF MESA/TESE SPERMATOZOA IS ASSOCIATED WITH INCREASED FERTILIZATION AND IMPLANTATION RATES IN HUMAN

The key for human IVF using the intracytoplasmic sperm injection (ICSI) procedure is to find moving and living sperm in azoospermic men. The objective of this study was to determine the effect of induced sperm motility on the fertilization and implantation rates in humans. Testicular sperm extraction (TESE) medium containing sperm stimulators and nutritional elements is specially designed for male factor and/or poor quality sperm with lower motility. Men with obstructed or absent ejaculatory ducts were treated by using a microepididymal sperm aspiration (MESA) procedure. Sperm are aspirated directly from the epididymis for use in the ICSI procedure. Men with non-obstructive azoospermia were treated by using the TESE procedure. Small samples of testicular tissue are obtained by needle or microsurgical biopsy and a few sperm are painstakingly dissected out of the tissue for use in the ICSI procedure. Sperm specimens either from epididymis or testis were divided into the two respective groups. In Group I, the specimens were washed in a TESE medium plus 10% human serum albumin (HSA), and then incubated with TESE medium containing 10% HSA in a 5% CO2 incubator to induce sperm motility. In Group II, the specimens were washed in human tubal fluid (HTF) medium plus 10% HSA, and then incubated with HTF containing 10% HSA in a 5% CO2 incubator. After 1 h of incubation, the motility of spermatozoa was evaluated under a microscope. The ICSI procedure was performed under an inverted microscope. Fertilized eggs were cultured in HTF medium containing 10% serum substitute supplement (SSS) until Day 3 for embryo transfer. Patients' pregnancy was followed after embryo transfer. This study demonstrated that incubation of spermatozoa from MESA and TESE procedures in TESE medium, as compared with HTF medium, results in significantly increased sperm progression (MESA sperm: 2-3 vs. 0-1; TESE sperm: 1-1.5 vs. 0-0.1) and motility (MESA sperm: 12.0 � 1.8% vs. 3.3 � 0.5%; TESE sperm: 4.5 � 0.2% vs. 0.15 � 0.02%) all respectively. Induced motility of spermatozoa in TESE medium, as compared with HTF medium, is associated with a significantly increased fertilization rate (MESA sperm: 74.5 � 3.5% vs. 60.2 � 3.1; TESE sperm: 73 � 4.3% vs. 48.4 � 3.7%) and implantation rate (MESA sperm: 29.8 � 3.8% vs. 19.4 � 2.7%, for patient age 32 � 3.0, in Group I, n = 13, and in Group II, n = 12; TESE sperm: 23.2 � 3.3% vs. 15.4 � 2.8%, patient age 33 � 2.8%, in Group I, n = 11, and in Group II, n = 9), and significantly decreased the ICSI performing time (30 � 10 vs. 120 � 20 min), all respectively. TESE medium can significantly increase the motility of spermatozoa from TESE/MESA, because it contains specific sperm stimulators and nutritional elements that directly resulted in the increase of fertilization and implantation rates.

PRONUCLEAR SYNCHRONIZATION AND NUCLEAR MORPHOLOGY OF MATURE AND IN VITRO MATURED OOCYTES IN THE RAT: AN ULTRASTRUCTURAL STUDY

The objective of this study was to evaluate synchronous and asynchronous pronucleus (PN) formation and the related patterns of juxtapositional nucleolus (n) formation in immature (prophase I [PI] and metaphase I [MI]) and mature (metaphase II [MII]) oocytes after fertilization, both ultrastructurally and at the level of light microscope. A single dose of 15 IU gonadotrophin was injected subcutaneously to twenty four 26-wk-old, female Wistar rats to induce ovulation. Human chorionic gonadotrophin (4 IU) was administered 40 h later, and after 4-6 h the ovaries were dissected, and the oocytes were aspirated. A total of 214 rat oocytes were classified according to a maturation index as follows: group I, 80 PI oocytes; group II, 50 MI oocytes; and group III, 84 MII oocytes. Immature oocytes were in vitro matured for 18-36 h. Spermatozoa were acquired by microepididymal sperm aspiration and processed using swim-up technique. Intracytoplasmic sperm injection was performed on mature oocytes after 2 h of incubation and on in vitro matured (IVM) oocytes 4 h after maturation. Pronuclear synchronization [both pronucleases (PNs) centrally located, equal sized, with equal numbers and sizes of juxtapositional nucleoli (Nn)] was observed in fertilized oocytes. Asynchronous PN formation (diversity between male and female PNs, related to dimensions, localization, and the number of Nn) in groups I, II, and III was found in 75, 86, and 47% of preembryos, respectively. There was a significant difference of synchronous pronuclear formation between mature and IVM oocytes (P < 0.05). In IVM oocytes, asynchronous PN formation is high, and juxtapositional pronucleolar patterns are observed to be low by transmission electron microscope (TEM).

The Efficacy of Repeat Percutaneous Epididymal Sperm Aspiration Procedures

The development of intracytoplasmic sperm injection spawned new methods of sperm retrieval for men with obstructive azoospermia who did not want to undergo reconstruction of the seminal tract. There is a wide array of different procedures that may be performed in these cases, for instance percutaneous epididymal sperm aspiration (PESA) and microepididymal sperm aspiration. However, concerns regarding the presence of sperm in a second PESA attempt due to possible fibrosis have been suggested by many authors. We evaluate if it is worthwhile to repeat percutaneous epididymal sperm aspiration. The records of 20 patients (23 attempts) who underwent repeat PESA from January 1996 to September 2000 for assisted reproductive technique purposes were reviewed. In all patients the repeat procedure was performed on the same side as the previous PESA. Data were collected on patient age, presence of motile sperm during PESA, epididymal side, pregnancies and abortion rates. Mean patient age +/- SD was 32.4 +/- 5.6 years. One patient was excluded from our analysis due to lack of information on the chart regarding the side of the procedure. Repeat PESA was performed in the right epididymis in 12 attempts and in the left in 10. Of the remaining 19 patients 14 (73.68%) did not and 5 (26.3%) have sperm in the epididymal fluid. In these 5 patients 8 repeat PESA procedures were performed (3 procedures in 1 and motile sperm was always found (8 of 22 attempts, 36.4%). Three patients achieved pregnancy with the motile sperm retrieved from the repeat PESA (3 of 8 repeat attempts, 37.5%). No abortions were detected. More than a third of repeat PESA attempts resulted in the presence of motile sperm. Before performing testicular sperm aspiration or extraction in patients who have undergone previous PESA without achieving pregnancy, repeat PESA may be done. Further attempts should be added in the future to confirm these results.

Percutaneous Testicular Sperm Aspiration and Intracytoplasmic Sperm Injection in Obstructive and Non-Obstructive Azoospermia: An Easy Alternative to TESE and MESA

Objective: To evaluate the recovery rate of sperm from the testis using percutaneous testicular aspiration with a 22-gauge hypodermic needle followed by evaluation of the fertilization rate and pregnancy rate after intracytoplasmic sperm injection. Materials and Methods: This is a prospective observational study performed in a private in vitro fertilization setting in Kuwait. Fifteen patients with obstructive and non-obstructive azoospermia were included in the study. Thirteen of them had previous microepididymal sperm aspiration, percutaneous epididymal sperm aspiration or testicular sperm extraction. The sperm were retrieved using percutaneous testicular aspiration under local analgesia. This was followed by intracytoplasmic sperm injection. A total of 146 eggs were collected and 112 were injected. Results: Normal fertilization occurred in 91 oocytes (87.5%) and the total number of embryos cleaved was 83 (91%). Embryo transfer was performed in 13 with pregnancy rate of 33.3 per treatment cycle and 38.5 per embryo transfer. Failure to retrieve sperm was encountered in 2 cases both in the hypospermatogenesis group. Conclusions: Percutaneous testicular sperm aspiration using hypodermic needles under local analgesic is an easy and cheap method with high patient acceptability, minimal complications and no need of special training. In this small group, it seems to have an acceptable success rate in terms of sperm retrieval and pregnancy in the obstructive type as well as hypospermatogenesis, but to lesser extent.

Outcome of intracytoplasmic sperm injection in azoospermic patients: stressing the liaison between the urologist and reproductive medicine specialist

Objectives. To analyze the outcome of intracytoplasmic sperm injection (ICSI) cycles in infertile couples in whom the main diagnosis of infertility was azoospermia of obstructive and nonobstructive origin. Methods. Eighty-three consecutive ICSI cycles were carried out with retrieved testicular or epididymal spermatozoa, 60 cycles in 32 patients with obstructive azoospermia and 23 cycles in 12 patients with nonobstructive azoospermia. Fifty-four testicular biopsies (testicular sperm extraction) and 18 epididymal aspirations (microepididymal sperm aspiration) were performed. Results. Motile spermatozoa were recovered in 65 cycles (90.3%). In another 3 (4.2%), nonmotile spermatozoa were retrieved. In 4 patients (5.5%), sperm could not be recovered. In 11 cycles, frozen sperm from a previous procedure were used. A significantly lower fertilization rate (64% versus 73%, P = 0.02), clinical pregnancy rate (13% versus 47%, P <0.001), and good embryo quality rates (35% versus 56%, P = 0.009) were observed in patients with nonobstructive azoospermia. In patients with obstructive azoospermia, no significant differences were observed when the outcome was analyzed on the basis of the sperm origin (ie, from testicular sperm extraction or microepididymal sperm aspiration). Conclusions. When combining testicular sperm extraction or microepididymal sperm aspiration with ICSI in patients with obstructive azoospermia, the results in terms of fertilization, implantation, and pregnancy rates were similar to those found in patients with nonazoospermic obstruction who underwent ICSI with ejaculated sperm. Patients with nonobstructive azoospermia had lower fertilization, embryo quality, and pregnancy rates than did those with obstructive azoospermia, probably because of severe defects in spermatogenesis, leading to poor gamete quality. The urologist and reproductive endocrinologist now have an excellent therapeutic option to offer men with previously intractable infertility.

Comparison of the Fertilization Rates Between Motile and Immotile Spermatozoa from Frozen-Thawed Samples Derived from Microepididymal Sperm aspiration (MESA) and Testicular Sperm Extraction (TESE)

Objectives: A major problem in working with epididymal and testicular sperm is the difficulty in differentiating between live and dead sperm, because most of the sperm are immotile. The objectives of this study were (1) to determine if the phenomenon of sperm tail curling could be used as an indicator for the selection of viable sperm; and (2) to compare the fertilization rates between frozen-thawed motile and immotile spermatozoa obtained by testicular biopsy and epididymal sperm aspiration.

Correlation of a Clinical Selection Protocol with Testicular Pathology in Men with Azoospermia Undergoing Testicular Sperm Extraction

Objective: Azoospermia is common in the infertile men population. It is almost always due to defective spermatogenesis or genital tract obstruction. Men with obstructive azoospermia are suitable for PESA (percutaneous sperm aspiration), MESA (microepididymal sperm aspiration) or TESA (testicular sperm aspiration) because they usually have normal spermatogenesis. It is easier to process epididymal spermatozoa compared with testicular ones. On the other hand, sperm production is sparsely distributed in azoospermic men with defective spermatogenesis, making TESE (testicular sperm extraction) the best method of sperm retrieval. Accurate distinction between defective spermatogenesis and genital tract obstructive in men with azoospermia has been based on the use of genital tract exploration, testicular biopsy and histology but these procedures are invasive. The objective of this study was to assess the value of a clinical protocol in selecting azoospermic patients with defective spermatogenesis for testicular sperm extraction.

The first births and ongoing pregnancies associated with sperm cryopreservation within evacuated egg zonae.

This new procedure principally aims to avoid a second or possibly multiple surgical procedures for sperm extraction from the male partner in cases of limited amounts of sperm cells, where normal freeze-thaw protocols would fail. Patients (n = 34) diagnosed as azoospermic, extreme oligozoospermic, or oligoasthenozoospermic underwent the process of sperm cryopreservation within evacuated egg zonae. Other samples were allocated to conventional sperm freezing. Sperm samples were acquired using testicular sperm extraction (TESE), microepididymal sperm aspiration (MESA), or fresh ejaculate. Subsequently, five of these 34 couples have undergone in-vitro fertilization (IVF) and achieved normal fertilization using post-thawed spermatozoa frozen under zonae pellucidae in conjunction with intracytoplasmic sperm injection (ICSI). The average fertilization rate for the post-thaw injected spermatozoa was 65%. This is comparable with the regular fertilization rate of 65% for combined MESA and TESE using fresh spermatozoa. All patients underwent embryo transfer. The average implantation rate per embryo was 31%; nearly the same for regular MESA/TESE ICSI cycles (32%). The first pregnancy associated with this procedure concluded with the full term delivery of healthy twin girls on July 18, 1997. The remaining four thaw procedures resulted in another twin delivery, an ongoing singleton gestation, a negative pregnancy test and a biochemical pregnancy respectively.

Cytogenetic abnormalities of unfertilized oocytes generated from in-vitro fertilization and intracytoplasmic sperm injection: a double-blind study.

In the present study we have assessed the cytogenetic abnormalities of unfertilized oocytes from in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) programmes during a one year period (July 1995 to July 1996) with the cytogenetic analysis being carried out in a double-blind manner. A total of 88 unfertilized ICSI and 85 unfertilized IVF oocytes were used for the study and of these 51 and 62 oocytes, in each respective group, were suitable for analysis. The haploidy, diploidy and aneuploidy rates between ICSI (62.7, 7.8 and 5.9%) and IVF (61.3, 9.7 and 14.5%) groups were similar. A significant inter-patient variation in the incidence of hypohaploidy was observed within the IVF group. Chromosomal fragmentation or breakage was observed at a similar rate in both groups of unfertilized oocytes (23.5 and 14.5% for ICSI and IVF respectively). A significantly higher proportion of ICSI oocytes contained sperm nuclei (27/51, 52.9%) than did IVF oocytes (20/62, 32.3%, P < 0.01). The distribution and state of sperm head chromatin in relation to oocyte chromosomal complement was studied in both groups. ICSI oocytes contained decondensed or swollen sperm nuclei in association with haploid oocyte chromosomes (12/27, 44.4%) or condensed sperm heads in oocytes showing no chromosomal complements (7/27, 25.9%). In IVF oocytes sperm heads were either arrested in the condensed state (5/20, 25%), metaphase stage (3/20, 15%) or had undergone premature chromosome condensation (PCC; 6/20, 30%) in association with haploid oocyte chromosomes. The incidence of PCC was similar in the two groups. A marked variation in the incidence of total chromosomal abnormality was observed between patients within both ICSI (0-75%) and IVF (0-71%) groups indicating a possible similarity in oocyte quality between the majority of male factor and tubal infertility patients. The type of sperm used in the two fertilization procedures showed an increased incidence of chromosomal breakage with ICSI-MESA (microepididymal sperm aspiration) spermatozoa (4/6, 67%) compared to the ICSI-ejaculated (6/35, 17.1%; P < 0.05), ICSI-testicular biopsy (2/10, 20%) and IVF-normospermic (9/62, 14.5%; P < 0.01) spermatozoa. Chromosomal fragmentation may be associated with the degree of difficulty experienced at sperm injection, especially with sperm retrieved from the reproductive tract. Thus chromosomal fragmentation in ICSI may need further investigation using a larger sample size in order to assess the possible causative factors.

Testicular sperm extraction and intracytoplasmic sperm injection: A simplified method for treatment of obstructive azoospermia

Objective: To assess the effectiveness of a simple method of testicular sperm extraction for the treatment of obstructive azoospermia. Design: Retrospective study. Setting: Teaching tertiary medical center. Patient(s): Seventeen men with obstructive azoospermia. Intervention(s): The patients underwent 19 cycles of intracytoplasmic sperm injection (ICSI) using testicular sperm. In 5 cycles, testicular sperm extraction was performed after failed microepididymal sperm aspiration. In 14 cycles, testicular sperm extraction was performed in the office under local anesthesia from the outset. The outcome was compared with ICSI cycles using ejaculated sperm (95 cycles) and epididymal sperm (12 cycles fresh and 9 cycles frozen-thawed). Main Outcome Measure(s): Clinical pregnancy and implantation rates. Result(s): There were no differences in the fertilization, cleavage, implantation, or clinical pregnancy rates among ICSI cycles using testicular, epididymal (fresh or frozen-thawed), or ejaculated sperm. Conclusion(s): When used in conjunction with ICSI, testicular sperm extraction from small excisional biopsy is a simple and cost-effective method for the treatment of obstructive azoospermia.

Approaches for obtaining sperm in patients with male factor infertility

Objective: To describe methods of sperm retrieval for intracytoplasmic sperm injection (ICSI) in patients with male factor infertility and to review the clinical results using sperm from the different sources. Design: The literature on sperm-obtaining methods and ICSI was reviewed. Studies related to this topic were identified through MEDLINE. Result(s): This review describes the evolution of sperm retrieval methods. Sperm can be obtained by microepididymal sperm aspiration (MESA), percutaneous sperm aspiration (PESA), and testicular sperm extraction (TESE),from patients with congenital absence of the vas deferens or acquired vas obstruction. When ICSI is performed with ejaculated, epididymal, or testicular sperm, good fertilization and pregnancy rates are achieved without significant differences among the various sperm sources. The original percutaneous sperm aspiration method has been modified slightly and yields successful results. Conclusion(s): Viable pregnancies can be achieved with ICSI by using not only ejaculated sperm, but also epididymal and testicular sperm. Microepididymal sperm aspiration, percutaneous sperm aspiration, modified percutaneous sperm aspiration, and testicular sperm extraction can be considered standard procedures to treat male factor infertility.

R-113. Testicular sperm extraction may be used if spermatozoa from micro-epididymal sperm aspiration were not useful

R-113. Testicular sperm extraction may be used if spermatozoa from micro-epididymal sperm aspiration were not useful

R-002. Efficacy and role of micro-epididymal sperm aspiration in unreconstructable obstruction of the reproductive tract

R-002. Efficacy and role of micro-epididymal sperm aspiration in unreconstructable obstruction of the reproductive tract

Testicular and epididymal sperm in a microinjection program: Methods of retrieval and results

To describe the methods of collection and laboratory preparation of epididymal and testicular sperm; to compare the fertilization and pregnancy rates; and to establish prognostic factors. Retrospective analysis. Academic reproductive medicine clinic. One hundred twelve consecutive microinjection cycles in 80 patients using either epididymal or testicular sperm. Sperm were collected by microepididymal sperm aspiration, open testicular biopsy, or fine needle tissue aspiration testicular biopsy. Fertilization rate, implantation, and clinical pregnancy rates. The fertilization rate was higher with epididymal sperm (67%) than with testicular sperm (50%). Implantation rates (fetal hearts per embryo, testicular: 11%, epididymal: 8%) and pregnancy rates (clinical pregnancy per oocyte collection procedure, testicular: 25%, epididymal: 29%) were not significantly different with epididymal and testicular sperm. Multiple regression analysis showed that normal fertilization rates were significantly lower with testicular sperm, immotile sperm, and severe spermatogenic disorders. Although fertilization rates are significantly lower with testicular sperm, higher implantation rates resulted in equivalent pregnancy rates. Thus, testicular aspiration of sperm for intracytoplasmic sperm injection is a simple, inexpensive method of sperm retrieval in cases of azoospermia resulting from genital tract obstruction or severe spermatogenic disorder.