Day Of Intracytoplasmic Sperm Injection Research Articles

MP43-02 COMBINATION OF MICROSCOPIC TESTICULAR SPERM EXTRACTION (MTESE) AND DENSITY GRADIENT PROCESSING TO OPTIMIZE SPERM RETRIEVAL AND PREGNANCY OUTCOMES IN MEN WITH NONOBSTRUCTIVE AZOOSPERMIA (NOA)

You have accessJournal of UrologyCME1 Apr 2023MP43-02 COMBINATION OF MICROSCOPIC TESTICULAR SPERM EXTRACTION (MTESE) AND DENSITY GRADIENT PROCESSING TO OPTIMIZE SPERM RETRIEVAL AND PREGNANCY OUTCOMES IN MEN WITH NONOBSTRUCTIVE AZOOSPERMIA (NOA) Nicholas Deebel, Banafsheh Nikmehr, Janmejay Hingu, Collette O'Connor, Rachel Ruble, Yueqiang Song, Ayse Vuruskan, Anuradha Devineni, Cihan Halicigil, Stuart Howards, Tamer Yalcinkaya, and Hooman Sadri-Ardekani Nicholas DeebelNicholas Deebel More articles by this author , Banafsheh NikmehrBanafsheh Nikmehr More articles by this author , Janmejay HinguJanmejay Hingu More articles by this author , Collette O'ConnorCollette O'Connor More articles by this author , Rachel RubleRachel Ruble More articles by this author , Yueqiang SongYueqiang Song More articles by this author , Ayse VuruskanAyse Vuruskan More articles by this author , Anuradha DevineniAnuradha Devineni More articles by this author , Cihan HalicigilCihan Halicigil More articles by this author , Stuart HowardsStuart Howards More articles by this author , Tamer YalcinkayaTamer Yalcinkaya More articles by this author , and Hooman Sadri-ArdekaniHooman Sadri-Ardekani More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003289.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: mTESE with Intracytoplasmic sperm injection (ICSI) is the gold standard for treating infertile couples when NOA in the male is present. However, using conventional tissue processing techniques, many patients will remain negative for sperm on mTESE. Standard practice in assisted reproduction technology (ART) does not utilize the density gradient centrifugation process following mTESE. We hypothesize that gradient utilization can enrich sperm retrieval following mTESE. METHODS: Men with NOA underwent mTESE from 2019-2022 by a single surgeon. The biopsy specimens underwent mechanical dissection. Subsequent tissue processing and gradient wash were carried out through a gradient column including 90% and 50% gradient solution diluted in standard sperm wash medium. Upon verification of sperm, the post-gradient sperm and remaining un-dissociated tissue were cryopreserved separately. On the day of ICSI, the frozen gradient sperm were thawed to recover viable sperm. If needed, frozen tissue was thawed, subjected to mechanical dissociation, and a subsequent gradient to obtain enough viable sperm. Fertilization and embryo development were assessed morphologically with the Embryoscope and genetically with preimplantation genetic testing for aneuploidy (PGT-A). Following embryo transfer, biochemical (ß-HCG) and clinical (gestational sac) pregnancy were assessed. RESULTS: Overall, 24 male patients (36±1.5 years old) with NOA underwent mTESE for an average operating time of 110.6±6.9 minutes (range: 37-177). The primary sperm identification rate after mechanical dissociation of tissue was 37.5% (n=9). The use of the density gradient led to an overall sperm retrieval rate of 70.8% (n=17) (p<0.05) and secondary sperm retrieval rate (sperm identification in initially negative patients) of 53.3% (n=8). Retrieved sperm were cryopreserved to be used for ICSI. Thirteen couples (average female age and AMH 32.1±1.6 and 3.2±0.8 ng/ml respectively) underwent ICSI using extracted sperm. The overall fertilization, per-cycle pregnancy, and per-transfer pregnancy rates were 66%, 76%, and 100% respectively. Amongst these couples, 6 underwent PGT-A, and 70.0% of tested blastocysts were euploid. CONCLUSIONS: Utilization of mTESE with subsequent gradient technology may serve as a highly sensitive sperm detection and enrichment method for NOA patients in order to obtain superior ART outcomes. Source of Funding: NA © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e601 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nicholas Deebel More articles by this author Banafsheh Nikmehr More articles by this author Janmejay Hingu More articles by this author Collette O'Connor More articles by this author Rachel Ruble More articles by this author Yueqiang Song More articles by this author Ayse Vuruskan More articles by this author Anuradha Devineni More articles by this author Cihan Halicigil More articles by this author Stuart Howards More articles by this author Tamer Yalcinkaya More articles by this author Hooman Sadri-Ardekani More articles by this author Expand All Advertisement PDF downloadLoading ...

O-150 Predicting the sperm retrieval of microsurgical testicular sperm extraction (mTESE) in infertile men with azoospermia on the day of TESE-ICSI

Abstract Study question Is the sperm retrieval rate of a small, pre-processed sample (PPS) of each TESE-biopsy representative for the sperm outcome on the day of ICSI? Summary answer The analysis of a PPS reliably reflects the probability of finding comparable numbers of sperm at time of TESE-ICSI. What is known already Azoospermia is defined as a condition where no spermatozoa are found in the ejaculate and is diagnosed in up to 15% among infertile men and in 11% of all patients attending our centre. The combination of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI) has become the standard treatment of azoospermic patients. However, no validated standard procedure has been identified to predict the exact sperm outcome of the cryopreserved TESE samples prior to TESE-ICSI so far. For optimal management of TESE-biopsies and the respective ICSI treatment, we developed a stepwise approach for the analysis of tissue samples. Study design, size, duration We retrospectively analysed the outcome of 872 microsurgically retrieved testicular biopsies of 198 patients of legal age who had a TESE-ICSI at our department between 2009 and 2019. From all 872 mTESE biopsies the number of sperm extracted from a small, pre-processed sample (PPS) before freezing procedure were known. The PPS was then compared to the number of sperm retrieved from the corresponding thawed specimen on the day of TESE-ICSI. Participants/materials, setting, methods During micro-TESE eight samples per testis are retrieved, then 1/10 of each biopsy is removed, digested with collagenase and screened for spermatozoa (pre-processed sample, PPS). If less than 100 spermatozoa are detected the absolute sperm number is recorded, otherwise the result is displayed as the maximum value of 100 sperm. On the day of ICSI, one or more TESE biopsies are thawed and processed for TESE-ICSI; the absolute sperm number is counted again. Main results and the role of chance Comparing the sperm yield of 872 TESE samples at time of ICSI to its respective PPS showed a similar sperm outcome with a minor deviation of ± 5 spermatozoa in 73.6% of all biopsies. However, 12.9% of the specimen had less and 13.4% had more spermatozoa. A negative sperm retrieval in the initial PPS was confirmed in 93.1% (268/288). PPS with 1-4 spermatozoa had a 27.2% (43/158) risk of complete absence of sperm on the day of ICSI, yet sperm detection (≥1 sperm) was positive in 72.8% (115/158) of the biopsies. With initially ≥5 spermatozoa present in the PPS, only 0.9% (4/426) of the biopsies had no sperm on the day of ICSI, vice versa 99.1% (422/426) were spermatozoa positive. A significant (p = 0.01) and strong (rs = 0.926) correlation of the sperm retrieval rates of the PPS and the ICSI sample was found meaning that the PPS reflects very well the sperm retrieval rate of the cryopreserved mTESE biopsy thawed at time of TESE-ICSI. However, if ≤ 4 sperm are found in the PPS, there is a relevant risk for a negative sperm retrieval on the day of ICSI and the couple should be carefully advised before start of treatment. Limitations, reasons for caution This analysis focussed on sperm prediction in cases of severe male factor infertility and therefore the sperm yield on the day of ICSI was chosen as primary outcome. The reproductive competence of the retrieved sperm in terms of pregnancy and birth rates should be subject to further investigation. Wider implications of the findings Treatment options for azoospermic patients are mostly related to the ability to find sperm on the day of ICSI. However, validated standards for sperm processing are missing. Therefore, a PPS seems to be a good option for prediction of sperm retrieval and improves counselling of the patients prior to TESE-ICSI. Trial registration number not applicable

Is a contralateral testicular exploration required at microdissection testicular sperm extraction for men with nonobstructive azoospermia, cryptozoospermia or severe oligozoospermia?

Men with nonobstructive azoospermia (NOA), cryptozoospermia and severe oligozoospermia are candidates for microdissection testicular sperm extraction (micro-TESE) and intracytoplasmic sperm injection (ICSI). We sought to evaluate micro-TESE outcomes and the need for bilateral testicular exploration in the three groups of men. We conducted a retrospective study of 233 consecutive micro-TESEs in men with nonobstructive azoospermia (n=173), cryptozoospermia (n=43) and severe oligozoospermia (n=17). The decision to terminate the micro-TESE after a unilateral or bilateral testicular exploration was determined at the time of surgery and was based on the presence or absence of mature spermatozoa in the harvested micro-biopsies. Final assessment of sperm recovery, on the day of ICSI, was reported as successful (available spermatozoon for ICSI) or unsuccessful (no spermatozoon for ICSI). Unilateral testicular exploration resulted in successful sperm retrieval in 43% (75/173), 79% (34/43) and 100% (17/17) of men with NOA, cryptozoospermia and severe oligozoospermia respectively. Therefore, 57%, 21% and none of the men with NOA, cryptozoospermia and severe oligozoospermia, respectively, required a bilateral micro-TESE. Overall, micro-TESE resulted in successful sperm retrieval in 52% (90/173), 91% (39/43) and 100% (17/17) of men with NOA, cryptozoospermia and severe oligozoospermia respectively. Our data indicate that men with severe oligozoospermia are unlikely to require a bilateral testicular exploration at micro-TESE. Moreover, most cryptozoospermic men will have a successful sperm retrieval by micro-TESE with the majority of these patients requiring a unilateral exploration. In contrast, over 50% of the men with nonobstructive azoospermia will require a bilateral micro-TESE.

PICSI vs. MACS for abnormal sperm DNA fragmentation ICSI cases: a prospective randomized trial.

To know which sperm selection technique, physiological intracytoplasmic sperm injection (PICSI) or magnetic-activated cell sorting (MACS), is better for the selection of sperm with abnormal sperm DNA fragmentation (SDF) in patients undergoing intracytoplasmic sperm injection (ICSI). A prospective randomized trial included 413 ICSI cases with abnormal SDF (> 20.3%) by TUNEL assay. Patients with at least 1 million total progressive motile sperm count were randomized to PICSI or MACS groups on the day of ICSI. PICSI depends on the hyaluronan binding of better SDF sperm where individual sperm was selected, while MACS selects non-apoptotic sperm population using Annexin V magnetic beads. All pre-implantation embryogenic parameters were observed and the main outcome was the ongoing pregnancy rate. There were no significant differences between patients allocated to PICSI and MACS in the studied parameters including pre-implantation embryological data, implantation, clinical pregnancy, and ongoing pregnancy rates. Meanwhile, sub-analysis according to the female age has shown that female patients with less than 30years of age in the MACS group had significantly higher good-quality blastocyst, clinical pregnancy, and ongoing pregnancy rates than the PICSI group. However, the higher implantation (p = 0.051), clinical pregnancy (p = 0.078), and ongoing pregnancy (p = 0.097) rates observed in females between 30 and 35years of age in the PICSI group did not reach significance level. PICSI and MACS are efficient techniques for sperm selection in cases with abnormal sperm DNA fragmentation. However, MACS is preferred when the females are younger than 30years, while PICSI is preferred in older females. NCT03398317 (retrospectively registered).

Sperm DNA fragmentation on the day of fertilization is not associated with embryologic or clinical outcomes after IVF/ICSI.

To evaluate if sperm DNA fragmentation (SDF) in the sample used for intracytoplasmic sperm injection (ICSI) impacts outcomes after euploid blastocyst transfer. Prospective cohort study of couples undergoing IVF with preimplantation genetic testing for aneuploidy from December 2014-June 2017. Sperm collected on the day of ICSI was analyzed for SDF using the sperm chromatin structure assay (SCSA®). Semen analysis parameters, embryologic outcomes, and clinical outcomes after euploid blastocyst transfer were compared between groups with DNA fragmentation index (DFI) ≤ 15% and DFI > 15% using Mann-Whitney U, t tests, and generalized linear mixed effects models. Two hundred thirty-four patients were included. One hundred seventy-nine men had DFI ≤ 15% (low DFI group) and 55 men had DFI > 15% group (high DFI group). Total motile sperm and sperm concentration were significantly lower in the group with DFI > 15% vs. DFI ≤ 15%. There was no difference in fertilization (86.3 vs. 84.2%, adjusted OR (95% CI) 0.86 (0.63-1.18)), blastulation (49.5 vs. 48.8%, adjusted OR 1.02 (0.75-1.36)), or euploidy (55.7 vs. 52.1%, adjusted OR 0.96 (0.7-1.31)) between the low and high DFI groups, respectively. Clinical outcomes were similar between low and high DFI groups, including implantation rate (68.8 vs. 79.8%), ongoing pregnancy rate (65.9 vs. 72.6%), and miscarriage rate (4.2 vs. 8.8%), respectively. Sperm DNA fragmentation on the day of ICSI is not associated with embryologic or clinical outcomes after euploid blastocyst transfer. Increasing levels of SDF are associated with low sperm concentration and total motile sperm count.

A point of confusion for embryologists in the identification of viable spermatozoa by the eosin-nigrosin test.

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients’ semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.

88 EFFECT OF BICARBONATE/CO2 LEVEL DURING EMBRYO CULTURE ON EQUINE BLASTOCYST RATE AFTER INTRACYTOPLASMIC SPERM INJECTION

Equine embryos appear to require a high glucose concentration for development to the blastocyst stage. The complete cell-culture medium, DMEM/F-12 (DM), which contains 17 mM glucose, has been widely used for equine embryo culture; however, in other species, high glucose during the early stages of embryo development is detrimental. To avoid this, we initiated a 2-step system using a low-glucose human embryo culture medium (Global) from Days 0 to 5 [Day 0 = day of intracytoplasmic sperm injection (ICSI)], with glucose (20 mM) added to the medium in the second step (Days 5 to 10; Choi et al. 2015 Reproduction 150, 31–41). We noted a high pregnancy loss rate (20%) in our clinical ICSI program (Hinrichs et al. 2014 J. Equine Vet. Sci. 34, 176), which used this 2-step Global system. Limited data are available on pregnancy with DM-produced embryos, but in one study, the loss rate was 1/13 (7.7%; Choi et al. 2011 Reproduction 142, 529–538). It is possible that use of DM in the second step of culture would better support normal blastocyst development than does Global with added glucose. However, DM is typically used at 5% CO2, and Global at 6% CO2, so use of both media would necessitate 2 sets of incubators. In the present study, we explored the use of DM in the second step of a two-step equine embryo culture system, under different CO2 environments. Oocytes were collected from research mares via follicle aspiration and were held overnight before being matured in vitro for 30 h. All media included 10% fetal bovine serum. On Days 0 to 5 after ICSI, all embryos were cultured in Global under 6% CO2 in mixed gas (5% O2 and remainder N2) at 38.2°C. In Experiment 1, on Day 5, embryos were transferred to DM prepared according to our standard method, with 14.3 mM NaHCO3 and 5 mM NaOH, and were cultured in mixed gas at either 5% CO2 or 6% CO2. Five replicates were performed. In Experiment 2, DM was prepared by our standard method, or with 24.2 mM bicarbonate and no NaOH. When pH was measured using a pH meter after media were equilibrated overnight, this higher bicarbonate provided the same pH at 6% CO2 (pH 7.3), as was achieved with the standard DM preparation at 5% CO2. Six replicates were performed. In both experiments, blastocyst development was assessed on Days 7 to 10, and blastocyst rates were compared between treatments by Fisher’s exact test. In Experiment 1, blastocyst rates were 43%, 13/30 and 27%, 8/30 for the standard DM preparation in 5% and 6% CO2, respectively (P &gt; 0.05). In Experiment 2, the blastocyst rates were 34%, 14/44 for the standard DM preparation at 5% CO2 and 43%, 19/44 for the high-bicarbonate DM at 6% CO2 (P &gt; 0.05). We conclude that a 2-step Global-DM system can support equine blastocyst production under a consistent CO2 environment (6%) if DM bicarbonate levels are adjusted to balance the increased CO2. This work was supported by the Clinical Equine ICSI Program, Texas A&amp;M University, and by the Link Equine Research Endowment Fund, Texas A&amp;M University.

PD05-10 ICSI OUTCOME IN PATIENTS WITH HIGH DNA FRAGMENTATION: TESTICULAR VS EJACULATED SPERM: QATAR EXPERIENCE

You have accessJournal of UrologyInfertility: Therapy I1 Apr 2016PD05-10 ICSI OUTCOME IN PATIENTS WITH HIGH DNA FRAGMENTATION: TESTICULAR VS EJACULATED SPERM: QATAR EXPERIENCE Ahmad Hassan AlMalki, Haitham T ElBardisi, Sami S AlSaid, Hassan Burjaq, Moza AlBadr, and Mohamad M Arafa Ahmad Hassan AlMalkiAhmad Hassan AlMalki More articles by this author , Haitham T ElBardisiHaitham T ElBardisi More articles by this author , Sami S AlSaidSami S AlSaid More articles by this author , Hassan BurjaqHassan Burjaq More articles by this author , Moza AlBadrMoza AlBadr More articles by this author , and Mohamad M ArafaMohamad M Arafa More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2617AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Sperm DNA Fragmentation Index (DFI) has emerged as an important biomarker in the assessment of male fertility potential. There are many controversies involving the routine use of DFI in clinical evaluation of infertile male. Another controversy is using testicular sperm instead of ejaculated sperm aiming to improve assisted reproductive techniques outcomes. The aim of the present study was to evaluate Intra Cytoplasmic Sperm Injection (ICSI) outcomes in male patients with high DFI using Testicular Vs. Ejaculated Sperm. METHODS This is a prospective study of 38 men with high DFI not responding to treatment who were scheduled for ICSI between January 2014 to January 2015. Inclusion criteria were (1) High DFI (= 30% using HaloSperm) not responding to treatment (2) Previous ICSI trial using ejaculated sperm. Couples with female factor infertility were excluded. Routine semen analysis was done including sperm DFI. ICSI data of the previous cycle where ejaculated sperm were used for injection was documented. On the day of ICSI, retrieved oocytes were fertilized using testicular sperms obtained by TESA. Comparison of outcomes measures (oocytes fertilized, quality of embryos, clinical pregnancy rate and live birth rate) of the two fertilization techniques were documented. RESULTS Two patients were excluded from the study due to presence of female factor infertility. Mean male age was 38±12, while mean female age was 31±5. Semen Fluid Analysis (SFA) characteristics were as follow: Count 16±20; Motility (progressive) 19±20; Abnormal forms 86±16; Round cells (0.04±0.11)X10^6 and DFI was 56±15. Overall, there was no difference in the fertilization rate and embryo grading using ejaculate and testicular sperms (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% (5 of 36)). Also, 17 live births were documented in TESA group and only 3 live births were documents in ejaculate group (p<0.0001). CONCLUSIONS For infertile patients with High DFI = 30%, using testicular sperm for ICSI significantly increase clinical pregnancy rate as well as Live birth rate. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e172 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Ahmad Hassan AlMalki More articles by this author Haitham T ElBardisi More articles by this author Sami S AlSaid More articles by this author Hassan Burjaq More articles by this author Moza AlBadr More articles by this author Mohamad M Arafa More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

How to select immotile but viable spermatozoa on the day of intracytoplasmic sperm injection? An embryologist's view.

A viable spermatozoon is the prerequisite for initiating fertilization in intracytoplasmic sperm injection. Usually motility is the primary sign used to determine a sperm's viability. However, in every in vitro fertilization (IVF) laboratory cases are observed in which none or only few spermatozoa are motile. This can occur in treatment cycles involving ejaculated samples but is most common in cases where surgically extracted testicular spermatozoa are used. To aid in selection, several techniques have been developed to identify viable spermatozoa from the immotile fraction. Amongst the more commonly used approaches are (i) the hypo-osmotic swelling test (ii) chemical substances for induction of tail movements (iii) the sperm tail flexibility test and (iv) laser-assisted immotile sperm selection. All can be used routinely in an IVF laboratory with each having both strengths and weaknesses. It is the purpose of this short review to focus on the technical issues involved in the performance of each of these techniques and to highlight the advantages and disadvantages of each approach.

Correlation between adjusted late spermatid score and sperm count in normozoospermic and oligozoospermic men

Abstract Objective To correlate late spermatid score and sperm count in men with normal semen analysis and oligozoospermia undergoing intracytoplasmic sperm injection (ICSI) to set the average threshold of spermatids/tubule of testicular tissue needed for normal sperm count/ml of semen. Material and methods This study was conducted on 24 normozoospermic subjects and 18 oligozoospermic patients who underwent wide bore needle biopsy because of failed sperm collection at the day of ICSI. Clinical data were reviewed and testicular biopsy slides were examined for Johnson score and adjusted late spermatid score. Results The optimum cut off value for adjusted late spermatid score in predicting a sperm count >20 million/ml was 20.75 late spermatid per tubule with 100% specificity. Model equations were produced to predict the sperm count using adjusted late spermatid score and vice versa. Conclusion This work supports the hypothesis that late spermatid score is a simple and reliable method for quantitation of spermatogenesis and it correlates well with the sperm count. The value of late spermatids needed for normal sperm count in this report is different from some other reports. The better standardization of work in this study may help resetting a new late spermatid threshold for normal sperm count.

Ultrasonographic assessment of endometrial receptivity at embryo transfer in an in vitro maturation of oocyte program

Ultrasonographic assessment of endometrial receptivity at embryo transfer in an in vitro maturation of oocyte program

Ultrastructural analysis of TESE and semen sperm tails from patients exhibiting absolute immotility on the day of ICSI.

Sperm flagellar pathology was found to be the underlying cause of motility disorders that lead to male infertility. Conventional in vitro fertilization (IVF) procedures will fail when sperm show a total absence of motility. In such difficult cases intracytoplasmic sperm injection (ICSI) is the only available technique to fertilize an oocyte. Fertilization rates are low and may also be reduced when immotile sperm are used for ICSI from ejaculate of other than epididiymal or testicular origin. Presence of totally immotile sperm in the ejaculate on the day of ICSI if spermatogenesis is normal testicular sperm recovery can improve ICSI outcomes. But for patients having severe morphological or functional sperm defects embryos of lower quality tend to be produced when totally immotile sperm are used. In this study the 2 patients exhibiting totally immotile sperm in their ejaculates and TESE samples on the day of ICSI showed the same ultrastructural abnormalities. Peri-axonemal and axonemal abnormalities that were seen in association with sperm nucleus structural defects suggested that the source of sperm has no effect on morphologic characteristics and also reflects abnormality in both spermatogenesis and spermiogenesis. In this study the two patients who presented with oligoteratozoospermia with total immotility, using either ejaculate or TESE sperm fertilization and embryo development, can be obtained with ICSI, but no pregnancies were established after embryo transfers.