Sperm Extraction Research Articles

Comparative motility assessment of sperm retrieved from micro-testicular sperm extraction: A single-centre study comparing fresh and frozen-thawed sperm.

Microsurgical testicular sperm extraction (microTESE) is crucial for treating non-obstructive azoospermia (NOA), offering both 'fresh' and 'frozen' options. This study evaluates the impact of fresh versus frozen microTESE on the progression to intra-cytoplasmic sperm injection (ICSI) cycles, focusing on sperm motility. We conducted a retrospective analysis of microTESE procedures at a major medical centre from 2007 to 2021, excluding cases of obstructive azoospermia and cryptozoospermia. Patients were divided into two groups: fresh microTESE (Group FR) and frozen microTESE (Group FZ). Sperm motility was assessed, and ICSI outcomes were compared between groups. Out of 128 microTESE procedures on 113 NOA patients, 31 were fresh and 97 were frozen. Sperm was found in 67.7% of fresh cases and 45.3% of frozen cases. In fresh cases, 85.7% had motile sperm for ICSI, whereas in frozen cases, 81.8% had motile sperm initially, but only 52.7% retained motility post-thaw. Our findings indicate a significant drop in motile sperm availability for ICSI in frozen microTESE cases compared to fresh ones. This suggests a potential advantage of fresh microTESE for certain couples, despite the logistical challenges, highlighting the need for careful patient selection and counselling.

Role of Platelet-Rich Plasma in the Management of Non-obstructive Azoospermia.

Non-obstructive azoospermia (NOA) is a common cause of infertility in males, which is characterized by the absence of sperm in the ejaculate, resulting from impaired spermatogenesis. The primary therapeutic approaches for the management of NOA include testicular sperm extraction, varicocelectomy in case of clinical varicoceles, and hormonal manipulation. While traditional treatments are found to have a limited role in the management of NOA, recent studies have explored the potential of platelet-rich plasma (PRP) as a safer and promising therapeutic option. Autologous PRP preparations are derived from the patient's blood and comprise growth factors and cytokines, promoting tissue repair and regeneration. PRP has a wide range of applications in the medical field, including managing infertility in males and females. This literature review aimed to evaluate the existing evidence on the efficacy and safety of PRP in the management of NOA. After a thorough review of relevant data from observational and experimental studies, the findings of this study suggested that PRP may positively influence spermatogenesis, sperm quality, DNA integrity, and sperm retrieval during assisted reproductive procedures. Further research is needed to establish the optimal PRP preparation, administration method, and long-term benefits. The newer studies shall include a diverse patient population and employ long-term follow-up to assess the durability of any positive effects of PRP treatment. The growing body of evidence regarding the therapeutic potential for NOA in humans offers greater opportunities for men seeking fertility treatment, informing clinical practice, and optimizing the use of PRP.

The experience of transfeminine adolescents and their parents regarding fertility preservation via testicular sperm extraction (TESE): a qualitative study.

How do transfeminine adolescents and their parents experience fertility preservation via testicular sperm extraction (TESE)? Participants experienced the TESE process positively when provided with appropriate guidance to navigate the decisional dilemma between preserving future biological parenthood and the pressure to start puberty suppression early. Sperm banking through ejaculation is not feasible for all transfeminine adolescents due to genital dysphoria and early puberty; for this group, TESE is the only alternative. However, during early puberty, they must postpone or pause puberty suppression until spermatogenesis is fully developed. All consecutive TESE patients in our centre and their parents were invited to participate. Between December 2022 and May 2023, we included 6 adolescents and 10 parents. We used a qualitative approach based on semi-structured interviews to study the experience of the transfeminine adolescents and their parents. Transfeminine adolescents diagnosed with gender dysphoria in early puberty (Tanner stage ≥2) who underwent TESE before the start of puberty suppression or gender-affirming hormones participated in this study. The interviews were recorded, manually transcribed and analysed using reflective thematic analysis focusing on understanding the participants' experiences of puberty and fertility preservation and the various factors and social processes influencing their decision to undergo TESE. Transfeminine adolescents decide to undergo invasive fertility preservation because of a possible, future desire for parenthood. Parents stressed the importance of preventing future decisional regret. They both saw masturbation to obtain a semen sample as unreachable. Even though TESE is a surgical procedure with possible complications and pain afterwards, this alternative seemed less dysphoric to them.Adolescents had to postpone the start of puberty suppression, waiting for spermatogenesis to complete. Given the unpredictable development of puberty, they felt a lot of stress and anxiety during this period and were primarily concerned about the lowering of their voice. Because of this puberty-related stress, they were constantly balancing their motivations for fertility preservation against the early start of puberty suppression to prevent unwanted irreversible pubertal body changes. To support this decisional dilemma, adolescents and their parents need adequate communication with each other and their healthcare providers to reflect on their stress for pubertal change. Furthermore, close, continuous surveillance of their current mental state by their parents, together with their psychologist and pubertal development by their physicians, is needed. Despite the challenging decision and postponement of puberty suppression, they retrospectively experienced it positively without any regret. All adolescents successfully underwent TESE, with viable spermatozoa obtained and without complications. This may result in a more positive reflective experience regarding fertility preservation. Furthermore, the perspectives of adolescents with the intention of TESE but who already started with puberty suppression before the preservation were not included. This study helps healthcare providers improve their counselling and guidance of transfeminine adolescents who consider postponing puberty suppression for fertility preservation via TESE. The adolescents and their parents know more clearly what to expect during fertility preservation. Specific peer-support programs may facilitate the decision process for upcoming adolescents. No external funding was used for this study. None of the authors have any conflicts of interest. Not applicable.

Sperm cryopreservation protocol for micro-TESE-retrieved sperm.

Azoospermia is characterized by the absence of sperm in the ejaculate and is categorized into obstructive azoospermia (OA) and nonobstructive azoospermia (NOA). For men with NOA, testicular sperm extraction (TESE) is the only method to obtain sperm for assisted reproductive technology (ART). Given the rarity of these sperm and the unpredictable success of subsequent retrieval attempts, cryopreservation of microdissection-TESE-obtained sperm is essential. Effective cryopreservation prevents the need for repeated surgical procedures and supports future ART attempts. After first delving into the physiological and molecular aspects of sperm cryopreservation, this review aims to examine the current methods and devices for preserving small numbers of sperm. It presents conventional freezing and vitrification techniques, evaluating their respective strengths and limitations in effectively preserving rare sperm, and compares the efficacy of using fresh versus cryopreserved testicular sperm.

Artificial intelligence interpretation of touch print smear cytology of testicular specimen from patients with azoospermia.

Identification of mature sperm at microdissection testicular sperm extraction (mTESE) is a crucial step of sperm retrieval to help patients with non-obstructive azoospermia (NOA) proceed to intracytoplasmic sperm injection. Touch print smear (TPS) cytology allows immediate interpretation and prompt sperm identification intraoperatively. In this study, we leverage machine learning (ML) to facilitate TPS reading and conquer the learning curve for new operators. One hundred seventy-six microscopic TPS images from the testicular specimen of patients with azoospermia at Taipei Veterans General Hospital were retrospectively collected, including categories of Sertoli cell, primary spermatocytes, round spermatids, elongated spermatids, immature sperm, and mature sperm. Among them, 118 images were assigned as the training set and 29 images as the validation set. RetinaNet (Lin et al. in IEEE Trans Pattern Anal Mach Intell. 42:318-327, 2020), a one-stage detection framework, was adopted for cell detection. The performance was evaluated at the cell level with average precision (AP) and recall, and the precision-recall (PR) curve was displayed among an independent testing set that contains 29 images that aim to assess the model. The training set consisted of 4772 annotated cells, including 1782 Sertoli cells, 314 primary spermatocytes, 443 round spermatids, 279 elongated spermatids, 504 immature sperm, and 1450 mature sperm. This study demonstrated the performance of each category and the overall AP and recall on the validation set, which were 80.47% and 96.69%. The overall AP and recall were 79.48% and 93.63% on the testing set, while increased to 85.29% and 93.80% once the post-meiotic cells were merged into one category. This study proposed an innovative approach that leveraged ML methods to facilitate the diagnosis of spermatogenesis at mTESE for patients with NOA. With the assistance of ML techniques, surgeons could determine the stages of spermatogenesis and provide timely histopathological diagnosis for infertile males.

Technologies to improve sperm retrieval in men undergoing micro-TESE for NOA.

Nonobstructive azoospermia (NOA) is considered the most challenging clinical scenario for infertile men and current treatments leave many men unsuccessful at being able to achieve a pregnancy with their partner using their own sperm. Microdissection testicular sperm extraction (micro-TESE) is the choice for men with NOA desiring to father children with their own gametes. Micro-TESE results in the highest numbers of sperm cells retrieved for use with in vitro fertilization/intracytoplasmic sperm injection. With suboptimal micro-TESE success rates of sperm retrieval and then pregnancy and live birth using the retrieved sperm with in vitro fertilization/intracytoplasmic sperm injection, advances to improve outcomes are necessary. This article comprehensively reviews the technologies investigated to date to improve the outcomes for men undergoing micro-TESE.

The effects of testicular stromal stem cells on surgically injured testicular tissue in rats.

This study investigated the effects of transplanted testicular stromal stem cells (tSSCs) on surgically damaged testis tissue. Ten-week-old male Wistar albino rats were divided into three groups: control (n = 6), damage (DG) (n = 6) and testicular stromal stem cell (TSSC) (n = 6) groups. Surgically induced damage was inflicted on the left testes of both the DG and TSSC groups, with no intervention on the right testes. In the TSSC group, damaged testes were treated with transplanted tSSCs, followed by orchiectomy after 15 days. Testes tissues were stained with haematoxylin-eosin (H&E), and recovery rates of functional structures were assessed by modified Johnsen scoring. The effects of tSSCs on testicular tissue were demonstrated by immunohistochemistry using BAX, BCL-2 and caspase 3. Serum testosterone levels were analysed using the enzyme-linked immunosorbent assay (ELISA) method. Surgical damage caused germ cell degeneration in some seminiferous tubules and a decrease in interstitial areas. With tSSC treatment, improvements in testicular architecture were identified through spermatogenesis in the seminiferous tubules and normal histological structures in the interstitial areas. Correspondingly, in the modified Johnsen score, the DG group showed a significant difference compared to the other groups (p = 0.001). High expressions of BAX, BCL-2 and caspase-3 in the DG group revealed prominent features of apoptosis. With the injection of tSSCs, these expressions significantly normalized according to H score analysis (all p = 0.004). Although serum testosterone levels in the tSSC group were higher compared to the control and DG groups, this difference was not statistically significant (p = 0.119). This study suggests transplanting tSSCs could accelerate tissue healing after testicular sperm extraction (TESE) surgery for azoospermia patients, potentially paving the way for a new and important clinical treatment.

The Efficacy of Aromatase Enzyme Inhibitor Administration before Microscopic Testicular Sperm Extraction (Micro TESE) in Men with Non-Obstructive Azoospermia

The Efficacy of Aromatase Enzyme Inhibitor Administration before Microscopic Testicular Sperm Extraction (Micro TESE) in Men with Non-Obstructive Azoospermia

Evaluation of the Results of Intracytoplasmic Sperm Injection and Microdissection Testicular Sperm Extraction Treatments in Patients with Nonobstructive Azoospermia According to Etiological Factors: A Retrospective Analysis

Objective: This study aimed to retrospectively compare the results of microdissection testicular sperm extraction (microTESE) and Intracytoplasmic sperm injection (ICSI) treatments in nonobstructive azoospermia (NOA) patients with different aetiologies. Determinants (clinical characteristics) for microTESE outcomes were compared between patients with successful sperm retrieval (SSR) and sperm retrieval failure (SRF). Methods: A total of 510 NOA patients who underwent microTESE between January 2015 and January 2024 were included in this study. Patients were classified according to the cause of NOA and SSR, fertilisation rate, clinical pregnancy, and overall live birth rate were evaluated. Results: The SSR rate was 44.1% in the whole population. The idiopathic patient group had the lowest SSR rate (X2: 34.81; p<0.01). There was no difference between the groups in terms of fertilisation rate, clinical pregnancy and overall live birth rate. There was a negative correlation between age and SSR rates in patients with idiopathic NOA (t:-0.27; p<0.01). SSR rates were higher in patients with cryptorchidism (right: t:0.8; P:0.003; left: t:0.72; p:0.002) and mumps orchitis (right: t:0.76; P<0.01; left: t:0.76; p=0.003). Conclusion: Etiology has a significant role in terms of SSR in patients with NOA. SSR was found to be significantly less in patients with idiopathic NOA compared to other causes. In addition, age and testicular volume were significant predictive factors for SSR in patients with idiopathic and acquired NOA.

Secondary azoospermia after a successful natural pregnancy: a primary prospective study

BackgroundTo date, there is a lack of studies conducted on males with secondary azoospermia as a potential cause of male infertility who had previously fathered children through natural conception. The current study aims to investigate the potential causes of secondary azoospermia as a presentation of male infertility as well as the prognostic factors that can impact sperm retrieval rate (SRR) while undergoing microdissection testicular sperm extraction (microTESE).ResultsThirty two patients were recruited from the andrology outpatient clinic from August 2023 till January 2024. The mean age of the patients was sixty-two years old. All patients had varicoceles. Twenty seven patients (84%) had palpable varicocele grade 2 and 3 on both sides. Further multivariate logistic regression analysis of the significant factors in the univariate regression revealed that younger age (OR 0.7, 95% C.I. 0.7-1.0, p = 0.03) and having a history of coronary artery disease (CAD) were predictable factors for negative TESE outcome (OR 123.1, 95% C.I. 3.2-4748.5, P = 0.01).ConclusionIt appears that the etiopathogenesis of secondary azoospermia are multifactorial. Varicocele and CAD are major factors to be considered. Future studies should be implemented deploying larger pools of patients suffering from the same condition to affirm the findings of this primary study.

Testicular sperm extraction for fertility preservation in young patients with cancer.

Cancer survivors in the adolescent and young adult generation often experience marriage, pregnancy, and childbirth after treatment; thus, fertility preservation is very important. In male patients, testicular sperm extraction (TESE) is sometimes performed due to azoospermia. Such a procedure is called oncological TESE (onco-TESE). In the present study, we aimed to define onco-TESE as TESE for fertility preservation in cancer patients, including those receiving gonadotoxic treatment. Seventeen male patients with cancer who had undergone onco-TESE for fertility preservation at Yokohama City University Medical Center between April 2014 and March 2023 were included in the study. Motile testicular sperm were acquired by TESE in 9 out of 17 cases. Among patients who had initiated chemotherapy before surgery, Motile sperm could be acquired by onco-TESE in 3 out of 9 cases. In chemotherapy-naive patients, Motile sperm were acquired by onco-TESE in 6 out of 8 cases. In the end, sperm cryopreservation was performed in 10 patients. Cryopreserved sperm were used in 2 of the 10 cases, and live birth was achieved after intracytoplasmic sperm injection in both cases. Before starting gonadotoxic treatment, it is important to confirm whether the patient desires to bear children. If having a baby is desired, a referral to a reproductive medicine doctor is recommended. Fertility preservation before starting gonadotoxic treatment is preferable, but fertility preservation could be considered even after such a treatment.

MALDI Imaging Mass Spectrometry Reveals Lipid Alterations in Physiological and Sertoli Cell-Only Syndrome Human Testicular Tissue Sections.

Azoospermia, the absence of sperm cells in semen, affects around 15% of infertile males. Sertoli cell-only syndrome (SCOS) is the most common pathological lesion in the background of non-obstructive azoospermia and is characterised by the complete absence of germinal epithelium, with Sertoli cells exclusively present in the seminiferous tubules. Studies have shown a correlation between successful spermatogenesis and male fertility with lipid composition of spermatozoa, semen, seminal plasma or testis. The aim of this research was to discover the correlation between the Johnsen scoring system and phospholipid expressions in testicular cryosections of SCOS patients. MALDI imaging mass spectrometry is used to determine spatial distributions of molecular species, such as phospholipids. Phosphatidylcholines (PCs), phosphatidylethanolamines (PEs) and sphingomyelins (SMs) are the most abundant phospholipids in mammalian cells and testis. SMs, the structural components of plasma membranes, are crucial for spermatogenesis and sperm function. Plasmalogens, are unique PCs in testis with strong antioxidative properties. This study, using imaging mass spectrometry, demonstrates the local distribution of phospholipids, particularly SMs, PCs, plasmalogens and PEs in human testicular samples with SCOS for the first time. This study found a strong relationship between the Johnsen scoring system and phospholipid expression levels in human testicular tissues. Future findings could enable routine diagnostic techniques during microTESE procedures for successful sperm extraction.

Development of a deep-learning model for detecting positive tubules during sperm recovery for nonobstructive azoospermia.

To enhance surgical testicular sperm retrieval outcome for men with nonobstructive azoospermia, a deep-learning model was developed to identify positive seminiferous tubules by labeling 110 images with sperm-containing tubules sampled during microdissection testicular sperm extraction as training and validation data. After training, the model achieved an average precision of 0.60.

Clinical predictors of successful outcomes for couples with nonobstructive azoospermic male partners undergoing micro-TESE.

Nonobstructive azoospermia (NOA) is the most challenging and complex clinical scenario for infertile men. Besides circumstances such as hypogonadotropic hypogonadism, surgical sperm retrieval is typically necessary, and microdissection testicular sperm extraction (micro-TESE) is the procedure of choice for men with NOA desiring to father children with their own gametes. Micro-TESE results in the highest numbers of sperm cells retrieved for use with in vitro fertilization/intracytoplasmic sperm injection (ICSI) in comparison to all other techniques for surgical sperm retrieval in men with NOA. Several factors may affect sperm retrieval rate and ICSI outcomes, including the patient's age, testicular volume, histopathological and genetic profile, and serum hormone levels. This article aims to review the medical literature describing predictors of successful micro-TESE and the outcomes of ICSI in men with NOA.

Second chance in fertility: a comprehensive narrative review of redo micro-TESE outcomes after initial failure.

When microdissection testicular sperm extraction (micro-TESE) fails, a redo procedure may be the only option for patients who want a biological child. However, there are many gaps of knowledge surrounding the procedure, which need to be addressed to help clinicians and patients make informed decisions. This review explores redo micro-TESE in the context of nonobstructive azoospermia (NOA). Literature was searched using Google Scholar, Medline, and PubMed. Search terms were "NOA" AND "second microdissection testicular sperm extractions" AND "redo microdissection testicles sperm extraction" AND "repeat microdissection testicular sperm extractions" AND "failed microdissection testicular sperm extractions" AND "salvage microdissection testicular sperm extractions". Only original articles in English were included. A total of nine articles were included, consisting of four retrospective and five prospective studies. The time gap between the first and second micro-TESE varied from 6 months to 24 months. Most of the included studies reported successful surgical sperm retrieval (SSR) in the second micro-TESE in the range of 10%-21%, except in one study where it reached 42%. It has not been presented any definitive information about the use of hormonal treatment or the benefit of varicocelectomy prior to the second micro-TESE. Patients with hypospermatogenesis and Klinefelter syndrome (KS) had the highest chance of success in redo surgery. In conclusion, redo micro-TESE following a negative procedure can lead to sperm recovery in 10%-21%. Patients with hypospermatogenesis and KS have a higher chance of success. There is no enough evidence to conclude which is the best hormonal stimulation if any before a redo surgery.

Reproductive characteristics and success rate of intracytoplasmic sperm microinjection in spinal cord injury infertile men: Retrospective cohort study.

Thanks to modern methods of assisted reproduction (ART), parenthood has become an attainable goal for couples in which the male partner has experienced spinal cord injury (SCI). The aim of our study was to determine the success of the treatment of infertile patients with SCI with intracytoplasmic sperm injection (ICSI) of cryopreserved sperm obtained by the testicular sperm aspiration (TESA) procedure. In this retrospective study 156 infertile couples were included, in which the male partner is primarily infertile due to azoospermia. Infertile couples were divided into two groups. The first group (n= 82) includes men with SCI, and the second (n= 74) men with obstructive azoospermia (OA) as the cause of infertility. All infertile men were examined and processed in the diagnostic procedure, and based on the urological findings, surgical extraction of sperm from the testicles was indicated. Exclusion criteria were the age of women over 40 and men over 45. We found that the quality of sperm was worse in the group with SCI, compared to the group with OA, but without statistical significance. Zenica and Johnsen score (p= 0.001; p= 0.000) showed worse semen characteristics in the group with SCI. There were no significant differences in the average number of embryos (p= 0.698), pregnancy rates per cycle (p= 0.979) and pregnancy rates per embrio transfer (ET), clinical pregnancy rates per ET (p= 0.987) and delivery rates per ET (p= 0.804) in couples with SCI, compared to couples with OA. Based on the results of this research, the TESA and ICSI procedures can be recommended as a successful method in the treatment of male infertility caused by azoospermia due to SCI.

Improving Fertility in Non-obstructive Azoospermia: Results from an Autologous Bone Mar-row-Derived Mesenchymal Stromal/Stem Cell Phase I Clinical Trial.

In this phase I clinical trial, our primary objective was to develop an innovative therapeutic approach utilizing autologous bone marrow-derived mesenchymal stromal/stem cells (BM-MSCs) for the treatment of nonobstructive azoospermia (NOA). Additionally, we aimed to assess the feasibility and safety of this approach. We recruited 80 participants in this non-randomized, open-label clinical trial, including patients undergoing NOA treatment using autologous BM-MSCs (n=40) and those receiving hormone therapy as a control group (n=40). Detailed participant characteristics, such as age, baseline hormonal profiles, etiology of NOA, and medical history, were thoroughly documented. Autotransplantation of BM-MSCs into the testicular network was achieved using microsurgical testicular sperm extraction (microTESE). Semen analysis and hormonal assessments were performed both before and six months after treatment. Additionally, we conducted an in-silico analysis to explore potential protein-protein interactions between exosomes secreted from BM-MSCs and receptors present in human seminiferous tubule cells. Our results revealed significant improvements following treatment, including increased testosterone and inhibin B levels, elevated sperm concentration, and reduced levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and prolactin. Notably, in nine patients (22.5%) previously diagnosed with secondary infertility and exhibiting azoospermia before treatment, the proposed approach yielded successful outcomes, as indicated by hormonal profile changes over six months. Importantly, these improvements were achieved without complications. Additionally, our in-silico analysis identified potential binding interactions between the protein content of BM-MSC-derived exosomes and receptors integral to spermatogenesis. Autotransplantation of BM-MSCs into the testicular network using microTESE in NOA patients led to the regeneration of seminiferous tubules and the regulation of hormonal profiles governing spermatogenesis. Our findings support the safety and effectiveness of autologous BM-MSCs as a promising treatment modality for NOA, with a particular focus on the achieved outcomes in patients with secondary infertility (registration number: IRCT20190519043634N1).

Effect of time since vasectomy on live birth rate of TESE‒ICSI egg donation cycles and male-related prognostic factors.

Vasectomy is a widely used method of contraception. However, some men may have the desire to become biological fathers again after a period. To explore the effect of time since vasectomy and different male comorbidities on live birth rates from intracytoplasmic sperm injection cycles using donated oocytes by using testicular spermatozoa obtained by testicular sperm extraction. This was a retrospective study of 123 couples who underwent a testicular sperm extraction‒intracytoplasmic sperm injection cycle after vasectomy using donated oocytes. Subjects were divided into groups according to time since vasectomy and the male risk factor evaluated. The main outcomes measured were live birth rate per embryo transfer, per oocyte donation cycle, and per couple. We assessed the cumulative live birth rate according to the time since vasectomy and considered male comorbidities: body mass index, hypertension, diabetes mellitus, dyslipidemia, and smoking. The overall live birth rate per couple was 59.3% (50.6-68.0). Considering the number of embryo transfer and oocyte donation cycle, the live birth rates were 34.1% (27.8-40.4) and 44.5% (36.9-52.1), respectively. The live birth rate according to time since vasectomy was not statistically different between groups. Consequently, the cumulative live birth rate was similar between the different interval times when considering one to eight embryo transfers (p=0.74). No statistical differences in live birth rate and cumulative live birth rate were found between groups clustered according to male body mass index, smoking, hypertension, and dyslipidemia. However, diabetic male patients had a significantly lower rate of live birth rate per couple (22.2% [4.94-49.4]) than non-diabetic patients did (62.7% [53.7-71.8]) (p=0.03), but not in their cumulative live birth rate. The time since vasectomy seems to have no detrimental effects on the live birth rate and cumulative live birth rate in testicular sperm extraction‒intracytoplasmic sperm injection cycles with donated oocytes. Male diabetes negatively affects the overall live birth rate per couple, but not the cumulative live birth rate. These results could be useful for multidisciplinary patient-tailored counseling, regarding the chance of having a pregnancy and facilitating the decision-making process of the fertility specialists.

P-072 Fresh testicular sperm seems to yield more fertilization abnormalities and early pregnancy loss than frozen testicular sperm

Abstract Study question How do ICSI outcomes using fresh testicular sperm, compare to those using frozen samples cryopreserved from the previous fresh testicular sperm extraction (TESE)? Summary answer Although fresh testicular sperm led to a better fertilization and pregnancy rate,it yielded more fertilization abnormalities and miscarriages than frozen sperm for the same couples. What is known already Testicular biopsies can be performed in cases of severe male infertility to search for spermatozoa. In the current era of advanced reproductive techniques, ICSI in association with TESE may be an effective treatment for patients with azoospermia. TESE could be performed the day of oocyte pick-up, or instead, it may be accomplished previously, and in this case TESE spermatozoa are cryopreserved for the future ICSI cycles. However, when reviewing the literature, not a lot of papers tried to compare the ICSI outcomes between fresh and frozen testicular sperm in the same couples. Study design, size, duration This retrospective study included the clinical and embryological data of pairs of fresh and frozen sperm injection cycles from couples after TESE between 2010 and 2023. The first ICSI was performed with fresh spermatozoa retrieved from TESE. The subsequent ICSI cycles used frozen spermatozoa from the first TESE. Participants/materials, setting, methods All infertile couples that underwent ICSI+TESE with testicular sperm cryopreservation in the study period were identified (110 couples). Of whom, 34 couples who underwent 40 subsequent ICSI cycle using frozen spermatozoa from the previous TESE were included. ICSI cycles were divided in: Group 1:ICSI with TESE and fresh testicular sperm injection Group 2:ICSI with frozen testicular sperm injection ICSI outcomes were compared for: fertilization/fertilization abnormalities rate, embryo quality, pregnancy and miscarriages rate, and live birth rate. Main results and the role of chance The average age of men who underwent ICSI was 42 years, while that of women was 33 years. On average, the couples had their ICSI with frozen sperm within 1.7 years after the TESE. There were no significant differences in women’s clinical parameters, the type of ovarian stimulation used nor its primary outcome between the two ICSI attempts. Although the primary outcome of ICSI showed a significantly higher fertilization rate for fresh testicular sperm compared to frozen sperm (64.4% vs 44.8%; p = 0.003), there was a tendency for more fertilization abnormalities when fresh testicular sperm was used (8.3% vs 6.2%, p = 0.4). Despite a better quality of cleavage embryos in ICSI with fresh testicular sperm (43.9% vs 39.2%, p = 0.6) and a higher pregnancy rate for the same group (21.9% vs 15.9%, p = 0.52), the miscarriage rate was twice as high when compared to the miscarriage rate of ICSI with frozen testicular sperm (6.7% vs 3.4%, p = 0.8), indicating that cryopreservation of testicular sperm could play a role in selecting better sperm for micro-injection. As for the live birth rate, ICSI with frozen sperm led to not statistically inferior rates when compared with ICSI with fresh sperm in the same couples (6.7% vs 10%, p = 0.6). Limitations, reasons for caution Due to our inclusion criteria, the study population may seem to be too reduced to reach significant results. To gain more informative insights, we recommend combining results from different centers and referring to the etiology of azoospermia that led to the TESE procedure. Wider implications of the findings Genetic testing and chromosomal aberration detection on testicular spermatozoa before and after cryopreservation could be proposed to compare the proportion of defected spermatozoa in both types of samples. Trial registration number not applicable

O-261 Frozen gametes differ in post-insemination dynamics to their fresh sibling counterparts in cycles using testicular sperm

Abstract Study question Are there differences in post-insemination events between fresh and frozen-derived gametes when testicular sperm extraction (TESE) is used? Summary answer Frozen-TESE leads to decreased fertilization, but not blastocyst euploidy rates; 2PN-lag time is significantly longer when frozen gametes are utilized compared to their fresh counterparts What is known already Studies indicate that the origin of testicular sperm influences the development potential of cleavage-stage embryos into blastocysts. Recent morphokinetic analysis suggest that embryos from frozen-TESE exhibit prolonged pronuclear stages and more frequent uneven cleavage compared to those from ejaculated sperm. Notably, no comparison with fresh-TESE was performed. To date, no studies have explored analysis in embryo morphokinetic parameters comparing fresh and frozen-TESE, especially when derived from the same patient.Such comparisons could provide a more accurate understanding of potential differences. Given that TESE occasionally involves the use of vitrified oocytes, it is essential to thoroughly examine outcomes across all combinations. Study design, size, duration Retrospective cohort study in a tertiary referral IVF centre including 72 cycles of 30 couples between January 2017 to September 2023. Following insemination by TESE-sperm, embryos were cultured in time-lapse (TL) incubator. Blastocysts ≥BL3CC (Gardner’s) underwent trophectoderm (TE) biopsy for Preimplantation Genetic Testing for aneuploidy (PGT-A) by next generation sequency (NGS). Participants/materials, setting, methods Gamete sources were fresh (fresh-TESE) or frozen-thawed-TESE (frozen-TESE) from the same patients with non-obstructive azoospermia (NOA), and fresh or vitrified (frozen) oocytes from the same female partner. Fertilization, usable blastocysts, and euploidy rates were recorded. Morphokinetics were annotated for time (t) of PB2 (tPB2), tPNa (PN-appearance), tPNf (PN-fading), t2-t9, CP (compaction), tM (Morula), tSB (start blastulation), tB (blastocyst) and timings between each developmental event. Main results and the role of chance Among included 980 oocyte/TESE dyads, 253 (25.8%) used fresh oocyte/sperm, 141 (14.4%) frozen oocyte/fresh sperm, 558 (56.9%) fresh oocyte/frozen sperm and 28 (2.9%) frozen oocyte/sperm. In oocyte/sperm dyads using fresh-TESE, normal fertilization (2PN) (52.0% vs. 44.5%) were higher compared to dyads using frozen-TESE (P < 0.01). Mixed-effects logistic-regression accounting for dependency structure between oocytes/sperm dyads of the same couple, showed an association of frozen-TESE with lower fertilization (OR: 0.64, 95%CI: 0.45-0.89, P = 0.008) and lower blastulation (³BL3CC)/MII (OR: 0.53, 95%CI: 0.37-0.77, P = 0.001), but similar blastulation (³BL3CC)/2PN (OR: 0.69, 95%CI: 0.45-1.07, P = 0.09). After accounting for female age, euploidy per tested blastocyst was similar whether fresh or frozen sperm was used (OR: 0.91, 95% CI: 0.46-1.79, P = 0.788). Frozen oocyte use on the other hand was not significantly associated with these outcomes (P > 0.05 for all). Analysis of morphokinetic parameters revealed that use of frozen dyads was associated with longer duration of 2PN after tPNa to tPNf: in frozen-TESE, 2.10h longer (95%CI: 0.55-3.64, P = 0.008); and in frozen oocytes, 2.34h longer (95%CI: 0.31-4.36, P = 0.024). Among embryos reaching blastulation, frozen-TESE embryos reached 6-cell stage earlier (tPNf to t6, 2.4h earlier, P = 0.009) but they were somewhat delayed in reaching expansion from there on (t6 to tEB, 4.4 hours later, P = 0.059). Limitations, reasons for caution In certain oocytes, insemination was conducted using either fresh or frozen immotile sperm that showed unresponsiveness to activation methods. However, the samples size hinders a comprehensive comparison. Wider implications of the findings Use of frozen-thawed TESE from NOA-patients may decrease fertilization and consequently lower blastocyst yield, but not euploidy rates. Synchronizing fresh-TESE with oocyte retrieval appears safer to maximize fertilization. Regardless of the gamete type, frozen-sourced sperm or oocyte impact 2PN-lag time, possibly indicating nuclear repair mechanisms post-freezing, requiring further investigation. Trial registration number 2310-ABU-020-VF