The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinc...The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages,with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup forrepeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providingpractitioners with a comprehensive analysis of each option’s strengths and limitations. Comparative studies indicate that, althoughfresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthawoutcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparablerates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular spermextraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and proceduralconvenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individualneeds and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes andprovide personalized reproductive care.展开更多
Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been sh...Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been shown that patients with sperm parameters abnormalities and abnormal morphology have a high rate of fragmentation and sperm DNA decondensation[2,3],and that sperm DNA fragmentation analysis could be used as a predictor factor of fertility potential[4].展开更多
The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm ...The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm fertilizing capacity.Among these,sperm concentration and motility are the first parameters to be evaluated through an estimation carried out by expert examiners.展开更多
Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in ...Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in antidepressant consumption.Antidepressants are associated with sexual dysfunction in both men and women;however,their role in male fertility has been scarcely studied.Fluoxetine and sertraline,two serotonin reuptake inhibitors(SSRIs),are among the most prescribed antidepressants worldwide.To determine their possible effects,human sperm cells were exposed to either sertraline or fluoxetine at concentrations previously found in blood and seminal fluid of patients undergoing treatment.Spermatozoa were incubated for up to 24 h at 37℃ and 5%CO_(2),and important functional parameters such as sperm motility,viability,mitochondrial membrane potential,cellular reactive oxygen species(ROS)production,chromatin/DNA integrity,acrosome status,and tyrosine phosphorylation were assessed.At low levels,fluoxetine consistently decreased progressive motility throughout time while promoting fluctuations in ROS levels and sperm capacitation.Nevertheless,it did not affect viability,mitochondrial membrane potential,acrosome reaction nor chromatin/DNA integrity.Sertraline,on the other hand,had little to nonsignificant impact at low doses,but affected almost all tested parameters at supratherapeutic concentrations.Altogether,our results suggest that both antidepressants may impair sperm function,possibly through different mechanisms of action,but fluoxetine is the only exhibiting mild negative effects at doses found in vivo.展开更多
Objective: To investigate the specific mechanism of hypoxia-inducible factor 1 alpha (HIF-1α) in the regulation of human sperm apoptosis, and to provide a new theoretical reference and scientific basis for the diagno...Objective: To investigate the specific mechanism of hypoxia-inducible factor 1 alpha (HIF-1α) in the regulation of human sperm apoptosis, and to provide a new theoretical reference and scientific basis for the diagnosis and treatment of asthenospermia and other related conditions. Methods: Semen samples were categorized into the normal group and asthenospermia group based on sperm motility criteria. HIF-1α interfering agent cobalt chloride (CoCl2) and guanylate cyclase activator (Lificiguat, YC-1) were added respectively, with a control group established accordingly. Sperm motility (using anterior viability rate as an index), apoptosis level, ATP level, mitochondrial membrane potential, and reactive oxygen species (ROS) level were measured. The expression levels of HIF-1α, p-PI3K, and Bcl-2 in the samples were analyzed using Western blotting. Results: Following CoCl2 treatment, there was a significant increase in sperm apoptosis compared to the normal control group (12.51% ± 2.50% VS 11.15% ± 2.42%);additionally, sperm motility (45.34% ± 3.37% VS 51.36% ± 11.68%), ATP production (11.51 ± 2.87 nM/µL VS 14.99 ± 2.83 nM/µL), ROS levels, and mitochondrial membrane potential all decreased significantly (all P α and p-PI3K increased significantly while Bcl-2 expression decreased (all P α in the YC-1 treatment group were decreased, and the expression level of Bcl-2 was increased (all P α can influence human sperm apoptosis and motility through the PI3K signaling pathway.展开更多
The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary p...The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary possibility of azoospermic men fathering their own genetic offspring.展开更多
High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,b...High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,but the 6th edition of the World Health Organization(WHO)manual for semen analysis included the SDF assessment in the chapter on extended semen examinations.展开更多
Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm D...Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm DNA.This oxidative damage is associated with male infertility.On the other hand,when RoS are produced at low levels,they participate in the redox signaling necessary for sperm capacitation.Capacitation-associated RoS are produced by the sperm oxidase,whose identity is still elusive,located in the plasma membrane of the spermatozoon.Ros,such as superoxide anion,hydrogen peroxide,nitric oxide,and peroxynitrite,activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events.Peroxiredoxins(PRDXs),antioxidant enzymes that fight against oxidative stress,regulate redox signaling during capacitation.Among them,PRDX6,which possesses peroxidase and calcium-independent phospholipase A,(iPLA,)activities,is the primary regulator of redox signaling and the antioxidant response in human spermatozoa.The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)pathway,and it is regulated by PRDX6 iPLA2,protein kinase C(PKC),and receptor-type protein tyrosine kinase.The understanding of redox signaling is crucial to pave theway fornovel diagnostic tools and treatments of male infertility.展开更多
Sperm cryopreservation is an essential technique for male fertility preservation,especially in men who are undergoing medical treatment.Conventional cryopreservation methods face limitations such as oxidative stress,D...Sperm cryopreservation is an essential technique for male fertility preservation,especially in men who are undergoing medical treatment.Conventional cryopreservation methods face limitations such as oxidative stress,DNA fragmentation,and cytotoxicity associated with traditional cryoprotectants like dimethyl sulfoxide(DMSO).Recent breakthroughs have focused on improving post-thaw sperm viability with novel cryoprotectants and innovative freezing strategies.Prospective approaches include the use of amino acid-based cryoprotectants,deep eutectic solvents,and antioxidants that have been described to prevent oxidative damage and maintain DNA integrity.Vitrification,a high-speed freezing technique that prevents ice crystal formation,has demonstrated superior outcomes compared to conventional slow freezing.Moreover,the Direct Dropping Method,a cryoprotectant-free approach,has been introduced as a contamination-minimizing technique that preserves sperm functionality.Multiomics tools are also utilized to determine biomarkers for protocol optimization.Despite these advancements,cryoprotectant toxicity is a central challenge,emphasizing the necessity for safer agents.Future research must focus on long-term sperm functionality and individualized cryopreservation strategies to maximize reproductive outcomes.The current review highlights the challenges associated with sperm cryopreservation,explores innovative strategies and novel cryoprotectants,underscores the significance of maintaining DNA integrity,and proposes future research directions to improve fertility preservation outcomes.展开更多
Medically assisted reproduction(MAR)techniques are highly dependent on the sperm quantity and quality.Low sperm concentrations can be bypassed at least to some point by the usage of more sophisticated MAR techniques l...Medically assisted reproduction(MAR)techniques are highly dependent on the sperm quantity and quality.Low sperm concentrations can be bypassed at least to some point by the usage of more sophisticated MAR techniques like intracytoplasmic sperm injection(ICSI).Compared to this,disruptions in established indicators of sperm quality like motility or morphology pose greater challenges for the therapy of couple infertility.展开更多
Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed a...Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed at the time of sperm retrieval.Onco-micro-TESE addresses the dual objectives of treating both infertility and the testicular tumor simultaneously.The technique is intricate,necessitating a comprehensive understanding of testicular anatomy,physiology,tumor biology,and advanced microsurgical methods.It aims to carefully extract viable spermatozoa while minimizing the risk of tumor dissemination.This review encapsulates the procedural intricacies,evaluates success determinants,including tumor pathology and spermatogenic tissue health,and discusses the implementation of imaging techniques for enhanced surgical precision.Ethical considerations are paramount,as the procedure implicates complex decision-making that weighs the potential oncological risks against the profound desire for fatherhood using the male gametes.The review aims to provide a holistic overview of onco-micro-TESE,detailing methodological advances,clinical outcomes,and the ethical landscape,thus offering an indispensable resource for clinicians navigating this multifaceted clinical scenario.展开更多
Background:Testicular sperm aspiration(TESA)is a minimally invasive testicular sperm retrieval technique that has been utilized in the treatment of male factor infertility.We sought to evaluate sperm retrieval outcome...Background:Testicular sperm aspiration(TESA)is a minimally invasive testicular sperm retrieval technique that has been utilized in the treatment of male factor infertility.We sought to evaluate sperm retrieval outcomes of primary and redo TESA in men with severe oligoasthenoteratozoospermia(OAT)and obstructive azoospermia(OA).Methods:This is a retrospective analysis of consecutive TESAs(primary and redo)for men with severe OAT and OA performed between January 2011 and August 2022 at a high-volume infertility center.We compared TESA outcomes in men with severe OAT to those with OA and compared outcomes of men who underwent primary and redo TESA on the same testicular unit.Results:439 TESAs(366 primary and 73 redo)in men with severe OAT(n=133)and OA(n=306)were included.Men with OA had significantly higher sperm retrieval rate(SRR)and motile SRR compared to men with severe OAT(99%vs.95%and 98%vs.83%,respectively,p<0.05).The requirement for multiple biopsies and the total number of aspirates were significantly lower in men with OA compared to those with severe OAT(15%vs.32%and 1.2±0.5 vs.1.4±0.7,respectively,p<0.05).In both groups,SRR,motile SRR,the requirement for multiple biopsies,and the total number of aspirates were not significantly different in primary compared to redo cases.Conclusion:Our data demonstrate that TESA retrieval rates are significantly higher in men with OA compared to those with severe OAT.The data also demonstrate that a redo TESA in these men is as effective as a primary TESA,suggesting that areas of active spermatogenesis are preserved 6 months after TESA.展开更多
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 onl...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.展开更多
Lymphomas represent one of the most common malignant diseases in young men and an important issue is how treatments will affect their reproductive health.It has been hypothesized that chemotherapies,similarly to envir...Lymphomas represent one of the most common malignant diseases in young men and an important issue is how treatments will affect their reproductive health.It has been hypothesized that chemotherapies,similarly to environmental chemicals,may alter the spermatogenic epigenome.Here,we report the genomic and epigenomic profiling of the sperm DNA from a 31-year-old Hodgkin lymphoma patient who faced recurrent spontaneous miscarriages in his couple 11-26 months after receiving chemotherapy with adriamycin,bleomycin,vinblastine,and dacarbazine(ABVD).In order to capture the potential deleterious impact of the ABVD treatment on mutational and methylation changes,we compared sperm DNA before and 26 months after chemotherapy with whole-genome sequencing(WGS)and reduced representation bisulfite sequencing(RRBS).The WGS analysis identified 403 variants following ABVD treatment,including 28 linked to genes crucial for embryogenesis.However,none were found in coding regions,indicating no impact of chemotherapy on protein function.The RRBS analysis identified 99high-quality differentially methylated regions(hqDMRs)for which methylation status changed upon chemotherapy.Those hqDRMs were associated with 87 differentially methylated genes,among which 14 are known to be important or expressed during embryo development.While no variants were detected in coding regions,promoter regions of several genes potentially important for embryo development contained variants or displayed an altered methylated status.These might in turn modify the corresponding gene expression and thus affect their function during key stages of embryogenesis,leading to potential developmental disorders or miscarriages.展开更多
Insulin-like growth factor 2(IGF2)is a critical endocrine mediator implicated in male reproductive physiology.To investigate the correlation between IGF2 protein levels and various aspects of male infertility,specific...Insulin-like growth factor 2(IGF2)is a critical endocrine mediator implicated in male reproductive physiology.To investigate the correlation between IGF2 protein levels and various aspects of male infertility,specifically focusing on sperm quality,inflammation,and DNA damage,a cohort of 320 male participants was recruited from the Women’s Hospital,Zhejiang University School of Medicine(Hangzhou,China)between 1^(st) January 2024 and 1^(st) March 2024.The relationship between IGF2 protein concentrations and sperm parameters was assessed,and Spearman correlation and linear regression analysis were employed to evaluate the independent associations between IGF2 protein levels and risk factors for infertility.Enzyme-linked immunosorbent assay(ELISA)was used to measure IGF2 protein levels in seminal plasma,alongside markers of inflammation(tumor necrosis factor-alpha[TNF-α]and interleukin-1β[IL-1β]).The relationship between seminal plasma IGF2 protein levels and DNA damage marker phosphorylated histone H2AX(γ-H2AX)was also explored.Our findings reveal that IGF2 protein expression decreased notably in patients with asthenospermia and teratospermia.Correlation analysis revealed nuanced associations between IGF2 protein levels and specific sperm parameters,and low IGF2 protein concentrations correlated with increased inflammation and DNA damage in sperm.The observed correlations between IGF2 protein levels and specific sperm parameters,along with its connection to inflammation and DNA damage,underscore the importance of IGF2 in the broader context of male reproductive health.These findings lay the groundwork for future research and potential therapeutic interventions targeting IGF2-related pathways to enhance male fertility.展开更多
Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial insemination.While freeze-th...Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial insemination.While freeze-thawing is known to impair sperm DNA integrity,whether the damage induced consists of single-(SSB)or double-strand breaks(DSB)has not been determined.In addition,no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions,or are rather spread throughout the regions linked to protamines.The main aim of the present work,therefore,was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in cattle.Results The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay,and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis(PFGE).Before cryopreservation,the incidence of SSB was 10.99%±4.62%and involved 20.56%±3.04%of sperm cells,whereas these figures significantly(P<0.0001)increased up to 34.11%±3.48%and 53.36%±11.00%in frozen-thawed sperm.In contrast,no significant differences in the incidence of DSB were observed(P>0.990)before and after cryopreservation(before:incidence of 13.91%±1.75%of sperm DNA affecting 56.04%±12.49%of sperm cells;after:incidence of 13.55%±1.55%of sperm DNA involving 53.36%±11.00%of sperm cells).Moreover,PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid(<31.5 kb)was greater(P<0.0001)after(27.00%±4.26%)than before freeze-thawing(15.57%±4.53%).These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines,which are structured in toroids.On the other hand,in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm(P=0.032 and P=0.005),but not with the size of the DNA fragments resulting from these breaks(P>0.05).Conclusion Cryopreservation of bovine sperm generates single-strand DNA breaks,which are mainly located in protamine-condensed toroidal regions.The incidence of DNA breaks in cryopreserved sperm has an impact on cat-tle fertility,regardless of the size of generated fragments.展开更多
According to the World Health Organization(WHO)manual,sperm concentration should be measured using an improved Neubauer hemocytometer,while sperm motility should be measured by manual assessment.However,in China,thous...According to the World Health Organization(WHO)manual,sperm concentration should be measured using an improved Neubauer hemocytometer,while sperm motility should be measured by manual assessment.However,in China,thousands of laboratories do not use the improved Neubauer hemocytometer or method;instead,the Makler counting chamber is one of the most widely used chambers.To study sources of error that could impact the measurement of the apparent concentration and motility of sperm using the Makler counting chamber and to verify its accuracy for clinical application,67 semen samples from patients attending the Department of Andrology,West China Second University Hospital,Sichuan University(Chengdu,China)between 13 September 2023 and 27 September 2023,were included.Compared with applying the cover glass immediately,delaying the application of the cover glass for 5 s,10 s,and 30 s resulted in average increases in the sperm concentration of 30.3%,74.1%,and 107.5%,respectively(all P<0.0001)and in the progressive motility(PR)of 17.7%,30.8%,and 39.6%,respectively(all P<0.0001).However,when the semen specimens were fixed with formaldehyde,a delay in the application of the cover glass for 5 s,10 s,and 30 s resulted in an average increase in the sperm concentration of 6.7%,10.8%,and 14.6%,respectively,compared with immediate application of the cover glass.The accumulation of motile sperm due to delays in the application of the cover glass is a significant source of error with the Makler counting chamber and should be avoided.展开更多
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 th...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 within 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.展开更多
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 mi...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.展开更多
To investigate the impact of preoperative serum follicle-stimulating hormone(FSH)levels on the probability of testicular sperm retrieval,we conducted a study of nonobstructive azoospermic(NOA)men with different testic...To investigate the impact of preoperative serum follicle-stimulating hormone(FSH)levels on the probability of testicular sperm retrieval,we conducted a study of nonobstructive azoospermic(NOA)men with different testicular volumes(TVs)who underwent microdissection testicular sperm extraction(micro-TESE).A total of 177 NOA patients undergoing micro-TESE for the first time from April 2019 to November 2022 in Shenzhen Zhongshan Obstetrics and Gynecology Hospital(formerly Shenzhen Zhongshan Urology Hospital,Shenzhen,China)were retrospectively reviewed.The subjects were divided into four groups based on average TV quartiles.Serum hormone levels in each TV group were compared between positive and negative sperm retrieval subgroups.Overall sperm retrieval rate was 57.6%.FSH levels(median[interquartile range])were higher in the positive sperm retrieval subgroup compared with the negative outcome subgroup when average TV was<5 ml(first quartile[Q1:TV<3 ml]:43.32[17.92]IU l^(−1) vs 32.95[18.56]IU l−1,P=0.048;second quartile[Q2:3 ml≤TV<5 ml]:31.31[15.37]IU l^(−1) vs 25.59[18.40]IU l^(−1),P=0.042).Elevated serum FSH levels were associated with successful micro-TESE sperm retrieval in NOA men whose average TVs were<5 ml(adjusted odds ratio[OR]:1.06 per unit increase;95%confidence interval[CI]:1.01–1.11;P=0.011).In men with TVs≥5 ml,larger TVs were associated with lower odds of sperm retrieval(adjusted OR:0.84 per 1 ml increase;95%CI:0.71–0.98;P=0.029).In conclusion,elevated serum FSH levels were associated with positive sperm retrieval in micro-TESE in NOA men with TVs<5 ml.In men with TV≥5 ml,increases in average TVs were associated with lower odds of sperm retrieval.展开更多
文摘The use of fresh versus frozen spermatozoa in men with nonobstructive azoospermia (NOA) undergoingin vitro fertilization (IVF)has been a debated hot topic among reproductive specialists. Each approach presents distinct advantages and disadvantages,with fresh sperm typically showing superior sperm quality, while frozen sperm offers logistical flexibility and a reliable backup forrepeated cycles. This review summarizes the latest advancements in sperm retrieval and cryopreservation techniques, providingpractitioners with a comprehensive analysis of each option’s strengths and limitations. Comparative studies indicate that, althoughfresh sperm often has better quality metrics, cryopreservation methods such as vitrification have significantly improved postthawoutcomes, making frozen sperm a viable choice in assisted reproductive technologies (ART). The findings show comparablerates for fertilization, implantation, clinical pregnancy, and live birth between fresh and frozen microdissection testicular spermextraction (micro-TESE) sperm in many cases, although patient-specific factors such as timing, cost-effectiveness, and proceduralconvenience should guide the final decision. Ultimately, the choice of using fresh or frozen sperm should align with the individualneeds and conditions of patients. This tailored approach, supported by the latest advancements, can optimize ART outcomes andprovide personalized reproductive care.
文摘Total or severe teratospermia affects the prognosis of fertility and causes serious problems for patients undergoing assisted reproduction[1].The pathophysiological mechanism of teratospermia is unclear.It has been shown that patients with sperm parameters abnormalities and abnormal morphology have a high rate of fragmentation and sperm DNA decondensation[2,3],and that sperm DNA fragmentation analysis could be used as a predictor factor of fertility potential[4].
文摘The analysis of the ejaculate,better known as spermiogram,represents the first and main step to identify whether a series of sperm quality parameters are within the norm and therefore are consistent with normal sperm fertilizing capacity.Among these,sperm concentration and motility are the first parameters to be evaluated through an estimation carried out by expert examiners.
文摘Depression currently affects about 280 million people worldwide and its prevalence has been increasing dramatically,especially among the young and people of reproductive age,which consequently leads to an increase in antidepressant consumption.Antidepressants are associated with sexual dysfunction in both men and women;however,their role in male fertility has been scarcely studied.Fluoxetine and sertraline,two serotonin reuptake inhibitors(SSRIs),are among the most prescribed antidepressants worldwide.To determine their possible effects,human sperm cells were exposed to either sertraline or fluoxetine at concentrations previously found in blood and seminal fluid of patients undergoing treatment.Spermatozoa were incubated for up to 24 h at 37℃ and 5%CO_(2),and important functional parameters such as sperm motility,viability,mitochondrial membrane potential,cellular reactive oxygen species(ROS)production,chromatin/DNA integrity,acrosome status,and tyrosine phosphorylation were assessed.At low levels,fluoxetine consistently decreased progressive motility throughout time while promoting fluctuations in ROS levels and sperm capacitation.Nevertheless,it did not affect viability,mitochondrial membrane potential,acrosome reaction nor chromatin/DNA integrity.Sertraline,on the other hand,had little to nonsignificant impact at low doses,but affected almost all tested parameters at supratherapeutic concentrations.Altogether,our results suggest that both antidepressants may impair sperm function,possibly through different mechanisms of action,but fluoxetine is the only exhibiting mild negative effects at doses found in vivo.
文摘Objective: To investigate the specific mechanism of hypoxia-inducible factor 1 alpha (HIF-1α) in the regulation of human sperm apoptosis, and to provide a new theoretical reference and scientific basis for the diagnosis and treatment of asthenospermia and other related conditions. Methods: Semen samples were categorized into the normal group and asthenospermia group based on sperm motility criteria. HIF-1α interfering agent cobalt chloride (CoCl2) and guanylate cyclase activator (Lificiguat, YC-1) were added respectively, with a control group established accordingly. Sperm motility (using anterior viability rate as an index), apoptosis level, ATP level, mitochondrial membrane potential, and reactive oxygen species (ROS) level were measured. The expression levels of HIF-1α, p-PI3K, and Bcl-2 in the samples were analyzed using Western blotting. Results: Following CoCl2 treatment, there was a significant increase in sperm apoptosis compared to the normal control group (12.51% ± 2.50% VS 11.15% ± 2.42%);additionally, sperm motility (45.34% ± 3.37% VS 51.36% ± 11.68%), ATP production (11.51 ± 2.87 nM/µL VS 14.99 ± 2.83 nM/µL), ROS levels, and mitochondrial membrane potential all decreased significantly (all P α and p-PI3K increased significantly while Bcl-2 expression decreased (all P α in the YC-1 treatment group were decreased, and the expression level of Bcl-2 was increased (all P α can influence human sperm apoptosis and motility through the PI3K signaling pathway.
文摘The advent of intracytoplasmic sperm injection,along with the realization that many men with azoospermia due to primary testicular failure may have a few spermatozoa in their testes,has resulted in the revolutionary possibility of azoospermic men fathering their own genetic offspring.
文摘High levels of sperm DNA fragmentation(SDF)are associated with reduced assisted reproductive technology(ART)outcomes.Currently,SDF is not included in routine clinical assessment of male partners of infertile couples,but the 6th edition of the World Health Organization(WHO)manual for semen analysis included the SDF assessment in the chapter on extended semen examinations.
基金supported by The Canadian Institutes of Health Research(PJT165962).
文摘Reactive oxygen species(Ros)play a dual role in mammalian spermatozoa.At high levels,they are detrimental to sperm function since they can promote oxidative stress that produces oxidation of protein,lipids,and sperm DNA.This oxidative damage is associated with male infertility.On the other hand,when RoS are produced at low levels,they participate in the redox signaling necessary for sperm capacitation.Capacitation-associated RoS are produced by the sperm oxidase,whose identity is still elusive,located in the plasma membrane of the spermatozoon.Ros,such as superoxide anion,hydrogen peroxide,nitric oxide,and peroxynitrite,activate protein kinases and inactivate protein phosphatases with the net increase of specific phosphorylation events.Peroxiredoxins(PRDXs),antioxidant enzymes that fight against oxidative stress,regulate redox signaling during capacitation.Among them,PRDX6,which possesses peroxidase and calcium-independent phospholipase A,(iPLA,)activities,is the primary regulator of redox signaling and the antioxidant response in human spermatozoa.The lysophosphatidic acid signaling is essential to maintain sperm viability by activating the phosphatidylinositol 3-kinase/protein kinase(PI3K/AKT)pathway,and it is regulated by PRDX6 iPLA2,protein kinase C(PKC),and receptor-type protein tyrosine kinase.The understanding of redox signaling is crucial to pave theway fornovel diagnostic tools and treatments of male infertility.
文摘Sperm cryopreservation is an essential technique for male fertility preservation,especially in men who are undergoing medical treatment.Conventional cryopreservation methods face limitations such as oxidative stress,DNA fragmentation,and cytotoxicity associated with traditional cryoprotectants like dimethyl sulfoxide(DMSO).Recent breakthroughs have focused on improving post-thaw sperm viability with novel cryoprotectants and innovative freezing strategies.Prospective approaches include the use of amino acid-based cryoprotectants,deep eutectic solvents,and antioxidants that have been described to prevent oxidative damage and maintain DNA integrity.Vitrification,a high-speed freezing technique that prevents ice crystal formation,has demonstrated superior outcomes compared to conventional slow freezing.Moreover,the Direct Dropping Method,a cryoprotectant-free approach,has been introduced as a contamination-minimizing technique that preserves sperm functionality.Multiomics tools are also utilized to determine biomarkers for protocol optimization.Despite these advancements,cryoprotectant toxicity is a central challenge,emphasizing the necessity for safer agents.Future research must focus on long-term sperm functionality and individualized cryopreservation strategies to maximize reproductive outcomes.The current review highlights the challenges associated with sperm cryopreservation,explores innovative strategies and novel cryoprotectants,underscores the significance of maintaining DNA integrity,and proposes future research directions to improve fertility preservation outcomes.
文摘Medically assisted reproduction(MAR)techniques are highly dependent on the sperm quantity and quality.Low sperm concentrations can be bypassed at least to some point by the usage of more sophisticated MAR techniques like intracytoplasmic sperm injection(ICSI).Compared to this,disruptions in established indicators of sperm quality like motility or morphology pose greater challenges for the therapy of couple infertility.
基金supported by the National Natural Science Foundation of China(No.82371633)Peking University Clinical Scientist Training Program and the Fundamental Research Funds for the Central University(BMU2023PYJ H012).
文摘Oncological microdissection testicular sperm extraction(onco-micro-TESE)represents a significant breakthrough for patients with nonobstructive azoospermia(NOA)and a concomitant in situ testicular tumor,to be managed at the time of sperm retrieval.Onco-micro-TESE addresses the dual objectives of treating both infertility and the testicular tumor simultaneously.The technique is intricate,necessitating a comprehensive understanding of testicular anatomy,physiology,tumor biology,and advanced microsurgical methods.It aims to carefully extract viable spermatozoa while minimizing the risk of tumor dissemination.This review encapsulates the procedural intricacies,evaluates success determinants,including tumor pathology and spermatogenic tissue health,and discusses the implementation of imaging techniques for enhanced surgical precision.Ethical considerations are paramount,as the procedure implicates complex decision-making that weighs the potential oncological risks against the profound desire for fatherhood using the male gametes.The review aims to provide a holistic overview of onco-micro-TESE,detailing methodological advances,clinical outcomes,and the ethical landscape,thus offering an indispensable resource for clinicians navigating this multifaceted clinical scenario.
文摘Background:Testicular sperm aspiration(TESA)is a minimally invasive testicular sperm retrieval technique that has been utilized in the treatment of male factor infertility.We sought to evaluate sperm retrieval outcomes of primary and redo TESA in men with severe oligoasthenoteratozoospermia(OAT)and obstructive azoospermia(OA).Methods:This is a retrospective analysis of consecutive TESAs(primary and redo)for men with severe OAT and OA performed between January 2011 and August 2022 at a high-volume infertility center.We compared TESA outcomes in men with severe OAT to those with OA and compared outcomes of men who underwent primary and redo TESA on the same testicular unit.Results:439 TESAs(366 primary and 73 redo)in men with severe OAT(n=133)and OA(n=306)were included.Men with OA had significantly higher sperm retrieval rate(SRR)and motile SRR compared to men with severe OAT(99%vs.95%and 98%vs.83%,respectively,p<0.05).The requirement for multiple biopsies and the total number of aspirates were significantly lower in men with OA compared to those with severe OAT(15%vs.32%and 1.2±0.5 vs.1.4±0.7,respectively,p<0.05).In both groups,SRR,motile SRR,the requirement for multiple biopsies,and the total number of aspirates were not significantly different in primary compared to redo cases.Conclusion:Our data demonstrate that TESA retrieval rates are significantly higher in men with OA compared to those with severe OAT.The data also demonstrate that a redo TESA in these men is as effective as a primary TESA,suggesting that areas of active spermatogenesis are preserved 6 months after TESA.
文摘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.
文摘Lymphomas represent one of the most common malignant diseases in young men and an important issue is how treatments will affect their reproductive health.It has been hypothesized that chemotherapies,similarly to environmental chemicals,may alter the spermatogenic epigenome.Here,we report the genomic and epigenomic profiling of the sperm DNA from a 31-year-old Hodgkin lymphoma patient who faced recurrent spontaneous miscarriages in his couple 11-26 months after receiving chemotherapy with adriamycin,bleomycin,vinblastine,and dacarbazine(ABVD).In order to capture the potential deleterious impact of the ABVD treatment on mutational and methylation changes,we compared sperm DNA before and 26 months after chemotherapy with whole-genome sequencing(WGS)and reduced representation bisulfite sequencing(RRBS).The WGS analysis identified 403 variants following ABVD treatment,including 28 linked to genes crucial for embryogenesis.However,none were found in coding regions,indicating no impact of chemotherapy on protein function.The RRBS analysis identified 99high-quality differentially methylated regions(hqDMRs)for which methylation status changed upon chemotherapy.Those hqDRMs were associated with 87 differentially methylated genes,among which 14 are known to be important or expressed during embryo development.While no variants were detected in coding regions,promoter regions of several genes potentially important for embryo development contained variants or displayed an altered methylated status.These might in turn modify the corresponding gene expression and thus affect their function during key stages of embryogenesis,leading to potential developmental disorders or miscarriages.
基金supported by grants from the National Natural Science Foundation of China(No.82370846)the Zhejiang Provincial Natural Science Foundation of China(No.LY24H070002).
文摘Insulin-like growth factor 2(IGF2)is a critical endocrine mediator implicated in male reproductive physiology.To investigate the correlation between IGF2 protein levels and various aspects of male infertility,specifically focusing on sperm quality,inflammation,and DNA damage,a cohort of 320 male participants was recruited from the Women’s Hospital,Zhejiang University School of Medicine(Hangzhou,China)between 1^(st) January 2024 and 1^(st) March 2024.The relationship between IGF2 protein concentrations and sperm parameters was assessed,and Spearman correlation and linear regression analysis were employed to evaluate the independent associations between IGF2 protein levels and risk factors for infertility.Enzyme-linked immunosorbent assay(ELISA)was used to measure IGF2 protein levels in seminal plasma,alongside markers of inflammation(tumor necrosis factor-alpha[TNF-α]and interleukin-1β[IL-1β]).The relationship between seminal plasma IGF2 protein levels and DNA damage marker phosphorylated histone H2AX(γ-H2AX)was also explored.Our findings reveal that IGF2 protein expression decreased notably in patients with asthenospermia and teratospermia.Correlation analysis revealed nuanced associations between IGF2 protein levels and specific sperm parameters,and low IGF2 protein concentrations correlated with increased inflammation and DNA damage in sperm.The observed correlations between IGF2 protein levels and specific sperm parameters,along with its connection to inflammation and DNA damage,underscore the importance of IGF2 in the broader context of male reproductive health.These findings lay the groundwork for future research and potential therapeutic interventions targeting IGF2-related pathways to enhance male fertility.
基金Ministry of Science,Innovation and Universities,Spain(NextGeneration EU fundsMaría Zambrano Program 124/MTAI/22+2 种基金and PID2020-113320RB-I00)Agency for Management of University and Research Grants,Regional Government of Catalonia,Spain(2021-SGR-00900)Catalan Institution for Research and Advanced Studies(ICREA).
文摘Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial insemination.While freeze-thawing is known to impair sperm DNA integrity,whether the damage induced consists of single-(SSB)or double-strand breaks(DSB)has not been determined.In addition,no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions,or are rather spread throughout the regions linked to protamines.The main aim of the present work,therefore,was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in cattle.Results The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay,and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis(PFGE).Before cryopreservation,the incidence of SSB was 10.99%±4.62%and involved 20.56%±3.04%of sperm cells,whereas these figures significantly(P<0.0001)increased up to 34.11%±3.48%and 53.36%±11.00%in frozen-thawed sperm.In contrast,no significant differences in the incidence of DSB were observed(P>0.990)before and after cryopreservation(before:incidence of 13.91%±1.75%of sperm DNA affecting 56.04%±12.49%of sperm cells;after:incidence of 13.55%±1.55%of sperm DNA involving 53.36%±11.00%of sperm cells).Moreover,PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid(<31.5 kb)was greater(P<0.0001)after(27.00%±4.26%)than before freeze-thawing(15.57%±4.53%).These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines,which are structured in toroids.On the other hand,in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm(P=0.032 and P=0.005),but not with the size of the DNA fragments resulting from these breaks(P>0.05).Conclusion Cryopreservation of bovine sperm generates single-strand DNA breaks,which are mainly located in protamine-condensed toroidal regions.The incidence of DNA breaks in cryopreserved sperm has an impact on cat-tle fertility,regardless of the size of generated fragments.
基金supported by the Natural Science Foundation of China(No.32171264 and No.81974226)the Sichuan Science and Technology Program(2023NSFSC1609)。
文摘According to the World Health Organization(WHO)manual,sperm concentration should be measured using an improved Neubauer hemocytometer,while sperm motility should be measured by manual assessment.However,in China,thousands of laboratories do not use the improved Neubauer hemocytometer or method;instead,the Makler counting chamber is one of the most widely used chambers.To study sources of error that could impact the measurement of the apparent concentration and motility of sperm using the Makler counting chamber and to verify its accuracy for clinical application,67 semen samples from patients attending the Department of Andrology,West China Second University Hospital,Sichuan University(Chengdu,China)between 13 September 2023 and 27 September 2023,were included.Compared with applying the cover glass immediately,delaying the application of the cover glass for 5 s,10 s,and 30 s resulted in average increases in the sperm concentration of 30.3%,74.1%,and 107.5%,respectively(all P<0.0001)and in the progressive motility(PR)of 17.7%,30.8%,and 39.6%,respectively(all P<0.0001).However,when the semen specimens were fixed with formaldehyde,a delay in the application of the cover glass for 5 s,10 s,and 30 s resulted in an average increase in the sperm concentration of 6.7%,10.8%,and 14.6%,respectively,compared with immediate application of the cover glass.The accumulation of motile sperm due to delays in the application of the cover glass is a significant source of error with the Makler counting chamber and should be avoided.
文摘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 within 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.
文摘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.
基金supported by the Shenzhen Fundamental Research Program(No.JCYJ20210324121807021).
文摘To investigate the impact of preoperative serum follicle-stimulating hormone(FSH)levels on the probability of testicular sperm retrieval,we conducted a study of nonobstructive azoospermic(NOA)men with different testicular volumes(TVs)who underwent microdissection testicular sperm extraction(micro-TESE).A total of 177 NOA patients undergoing micro-TESE for the first time from April 2019 to November 2022 in Shenzhen Zhongshan Obstetrics and Gynecology Hospital(formerly Shenzhen Zhongshan Urology Hospital,Shenzhen,China)were retrospectively reviewed.The subjects were divided into four groups based on average TV quartiles.Serum hormone levels in each TV group were compared between positive and negative sperm retrieval subgroups.Overall sperm retrieval rate was 57.6%.FSH levels(median[interquartile range])were higher in the positive sperm retrieval subgroup compared with the negative outcome subgroup when average TV was<5 ml(first quartile[Q1:TV<3 ml]:43.32[17.92]IU l^(−1) vs 32.95[18.56]IU l−1,P=0.048;second quartile[Q2:3 ml≤TV<5 ml]:31.31[15.37]IU l^(−1) vs 25.59[18.40]IU l^(−1),P=0.042).Elevated serum FSH levels were associated with successful micro-TESE sperm retrieval in NOA men whose average TVs were<5 ml(adjusted odds ratio[OR]:1.06 per unit increase;95%confidence interval[CI]:1.01–1.11;P=0.011).In men with TVs≥5 ml,larger TVs were associated with lower odds of sperm retrieval(adjusted OR:0.84 per 1 ml increase;95%CI:0.71–0.98;P=0.029).In conclusion,elevated serum FSH levels were associated with positive sperm retrieval in micro-TESE in NOA men with TVs<5 ml.In men with TV≥5 ml,increases in average TVs were associated with lower odds of sperm retrieval.
