Poor sperm quality and advancing age are associated with increased sperm DNA damage in infertile men

With increasing evidence for faulty paternal contribution to reproduction, there has been a steady increase in studies highlighting an association between sperm DNA damage, failed/delayed fertilisation and aberrant embryo development. Owing to prevailing ambiguity, the aims of the study were to analyse the genetic integrity of the male gamete and then to understand its association with age, standard semen parameters, lifestyle and occupational factors. The study included 504 subjects, attending university infertility clinic for fertility evaluation and treatment. Semen characteristics were analysed by standard criteria; terminal deoxynucelotidyl transferase-mediated nick end-labelling assay was employed for DNA damage assessment. The average incidence of sperm DNA damage in patients with normozoospermic semen parameters was <10%. Patients with oligozoospermia, severe oligozoospermia, oligoasthenoteratospermia, asthenoteratozoospermia and necrozoospermia had significantly higher level of sperm DNA damage (P < 0.001). Patients above 40 years of age had significantly high levels of DNA damage (P < 0.001) compared with their counterparts. Patients with varicocele and a history of alcohol consumption had higher incidence of spermatozoa with DNA damage (P < 0.01). Poor sperm characteristics in the ejaculate are associated with increased sperm DNA damage. Age-related increase in sperm DNA damage and association of the same with varicocele and alcohol consumption are also demonstrated.     

Famotidine does not affect human sperm fertility characteristics (motility,viability and DNA integrity) in vitro

Famotidine, a histamine‐2 receptor antagonist, is commonly used to relieve the acid‐related gastrointestinal diseases; however, its effect on human sperm parameters, and hence on sperm function, is still undetermined. Here, we intended to measure human sperm motility, viability, and DNA integrity of ejaculated human sperm in the presence of famotidine at 0, 0.1, 1 and 10 mM concentrations in vitro. Forty‐nine semen samples of normal count, motility, and morphology were included in this study. Sperm motility was assessed using Makler counting chamber and a phase contrast optics (200× magnification), whereas sperm viability was assessed using eosin–nigrosin staining procedure. The effect of famotidine on sperm DNA integrity was measured using flow cytometry. Famotidine at 0.1, 1 or 10 mM had insignificant effect on human sperm motility (progressive, p = .9594; and total, p = .8420), sperm viability (p = .6471), and content of DNA breaks in sperm (p > .05) compared with the control. In conclusion, famotidine at 0.1, 1 or 10 mM did not alter human sperm motility, viability or DNA integrity in vitro. Although, these findings indicate safety of famotidine in human sperm, further in vivo studies are required to establish the drug's safety.     

精子DNA碎片指数和精子畸形率对ICSI临床结局的影响

目的:通过评估ICSI治疗前的精子畸形率(SMR)和精子DNA碎片指数(DFI),探讨精子DFI和SMR对卵胞浆内单精子注射(ICSI)助孕结局的影响。方法:共入组79对因少弱精子症实施第一周期ICSI治疗的不孕夫妇,在进入治疗周期前3⁶个月,评价精子浓度、前向运动精子百分率、SMR及DFI。主要观察SMR和DFI与ICSI结局参数的关系。结果:79例少弱精子症患者DFI正常51例,异常28例,异常组的DFI值明显升高(14.18%vs 41.47%);巧合的是,SMR正常组同样为51例,异常组28例,异常组的SMR值亦明显升高(87.88%vs98.46%)。按DFI正常(DFI≤25%)与异常(DFI>25%)分组,或按SMR正常(≤96%)与异常(>96%)分组,组间的双方年龄、女方BMI、获卵数、移植胚胎数等基本情况差异无统计学意义。DFI正常和异常组间,SMR正常和异常组间的受精卵子数、可移植胚胎数、早期流产率无显著差异;异常组生化妊娠率(43.5%vs 61.5%)和临床妊娠率(39.1%vs 56.4%)降低,但差异无统计学意义(P=0.19及0.10)。精子DFI与SMR呈显著正相关(r=0.231,P<0.05)。结论:精子DFI增高(>25%),与按严格标准检测的SMR增高(>96%)男性行ICSI治疗,生化妊娠率和临床妊娠率降低,但与正常者比较未发现有统计学差异,可能与样本量小有关,有必要深入研究。     

Stability of sperm characteristics in men with disturbances in sperm quality

Sixty-one men referred to our laboratory for semen analysis, and subsequently judged to exhibit some form of sperm pathology, were asked to return for a second analysis, not less than 2 months after the first, in order to assess the stability of the pathological changes observed. In almost half of the cases, the referring physician had, on his own initiative, started hormone or antibiotic treatment. The sperm parameters studied included sperm count, sperm motility judged by laser-Doppler spectroscopy, and sperm morphology and viability. The motility characteristics included percentage motile, their average velocity, and percentage swimming in a progressive manner, and their progressive velocity. In untreated subjects, there was no significant difference between the first and second analysis in any of the sperm parameters measured. This was also true for both oligozoospermic individuals (less than 20 x 10(6) sperm/ml) and the group with higher sperm concentrations. All parameters were highly correlated on the two occasions. The average coefficients of variation of the paired observations were highest for sperm count (approximately 25%) and lowest for sperm velocities and the proportion of abnormal and viable cells in the ejaculate (1-9%). No major differences in the extent of variation could be detected between the low and high sperm density groups. In general, the unsystematic antibiotic and hormone regimens (clomiphene or androgen) used by the referring physicians had no discernable effect on any aspect of sperm quality, indicating the need for more controlled and standardized programmes of treatment.     

Localization of AKAP4 and tubulin proteins in sperm with reduced motility

Aim： To perform screening, related to A-kinase anchoring proteins 4 （AKAP4） and tubulin proteins, in spermatozoa with absent or severely reduced motility in order to detect the status of the fibrous sheath and the axonemal structure. Methods： An immunocytochemical study of tubulin, used as a positive control, and AKAP4 was carded out to detect the presence and the distribution of these proteins in different sperm samples. The morphological characteristics of sperm were studied by transmission electron microscope （TEM） and the results were elaborated using a formula reported in previous studies. PCR was carried out on DNA extracted from peripheral blood lymphocytes to analyse partial sequences of the Akap4 and Akap3 genes. Results： Immunolabelling of tubulin and AKAP4 showed different patterns, which led us to divide the patients into groups. In group I, the absence of AKAP4 and tubulin was revealed, although these patients did not show alterations in the Akap4/Akap3 binding site. TEM evaluation highlighted that a high presence of necrosis was associated with total sperm immotility. In group Ⅱ, a regular AKAP4 and tubulin signal was present, although motility was reduced and TEM analysis revealed the presence of immaturity. In group Ⅲ, in which a weak AKAP4 label associated with normal tubulin staining and reduced motility was observed, a severe disorganization of the fibrous sheath was highlighted by TEM. Conclusion： While the role of AKAP4 in sperm motility is unclear, absent or weak AKAP4-1abelling seems to be associated with absent or weak sperm motility.     

Human sperm DNA oxidation, motility and viability in the presence of L-carnitine during in vitro incubation and centrifugation

In vitro incubation and centrifugation is known to decrease human sperm quality. In the human body, besides its antioxidant effects, L-carnitine (LC) facilitates the transport of activated fatty acids from the cytosol to the mitochondrial matrix. In this study, we investigated the effect of LC on human sperm motility, viability and DNA oxidation after incubation and centrifugation, following the sperm preparation protocols of assisted reproduction. Normozoospermic semen samples (n = 55) were analysed according to the World Health Organization (WHO) guidelines. LC concentrations that are not toxic to spermatozoa as determined by sperm motility and viability were standardised after 2 and 4 h of incubation at 37 °C. Semen samples to which the optimal LC concentrations were added were also centrifuged for 20 min at 300 g and analysed for sperm motility, viability and DNA oxidation. Sperm motility was improved at 0.5 mg ml(-1) LC after incubation and centrifugation with 5 × 10(6) sperm ml(-1). Higher concentration of LC (50 mg ml(-1)) significantly decreased sperm motility and viability. LC did not alter the baseline of sperm DNA oxidation during both incubation and centrifugation. In conclusion, LC may enhance sperm motility following incubation and centrifugation, while it might not affect sperm viability and DNA oxidation.     

人精子体外获能前后计算机辅助分析参数的比较

目的为临床使用计算机辅助精子分析(CASA)系统的精液分析参数提供参考。方法对48例人类冷冻复温精液和116例临床进行丈夫人工授精(AIH)的新鲜精液体外获能前后CASA检测结果,分组进行配对资料t-检验和独立样本t-检验。结果所有实验组精子体外获能后的曲线运动速度(VCL)、直线运动速度(VSL)、平均路径速度(VAP)、精子头侧摆幅度(ALH)和鞭打频率(BCF)显著提高(P<0．01);除冻存精液精子体外获能后的运动前向性(STR)和头部椭圆度外,其余各组精液精子获能后的STR、直线性(LIN)和头部椭圆度显著下降(P<0,01)。行AIH治疗的精液标本,无论获能前后,已孕组精子VAP、LIN和头部椭圆度显著高于未孕组(P<0．05),而BCF和头部面积低于未孕组(P<0.05);获能前两组的(a b)级精子百分率无显著性差异,获能后差异显著(P<0．05)。结论体外获能处理可显著提高精子的各种运动速度,但精子运动的前向性和直线性降低;精液常规分析结果中的精子运动等级和速度不直接反映精子的受精能力。     

Relationship between sperm quality and chromatin condensation measured by sperm DNA fluorescence using flow cytometry

DNA flow cytometry of sperm from 100 randomly chosen men undergoing fertility investigation revealed a general association between reduced sperm quality, as judged by conventional parameters, and the appearance of sperm with lower degrees of chromatin condensation in the ejaculate as measured by DNA fluorescence intensity. Chromatin hypocondensation, as measured by increased fluorescence, was manifested to different degrees in different samples. In many cases of more extreme sperm pathology, such as oligoasthenoteratozoospermia (OAT), the whole population of spermatozoa appeared to be affected. Significant numbers of hypercondensed spermatozoa were present in both normozoospermic men and men with different degrees of disturbance in sperm quality. All of the different parameters of sperm quality could be correlated significantly with certain of the flow parameters, although not one in particular could be used to predict deviations from the normal flow profile. In several asthenoteratozoospermic men and a small proportion of men with OAT, the DNA profiles were normal, implying that in these cases the disturbance may not be so fundamental. The presence of leucocytes in the ejaculate was associated with a general increase in the preponderance of hypocondensed subpopulations of spermatozoa in men with OAT as well as in normozoospermic subjects, emphasizing the effect of inflammatory conditions in the reproductive tract on sperm quality.     

Impact of cigarette‐smoking on sperm DNA methylation and its effect on sperm parameters

DNA methylation is an epigenetic modification of the genome. The purpose of this study was to determine the influence of cigarette‐smoking on sperm DNA methylation from a genomewide survey of sperm samples and to ascertain its effect on sperm parameters. Twenty‐eight sperm DNA samples (from 14 fertile smokers as a case study and 14 proven fertile nonsmokers as controls) were subjected to Infinium 450K BeadChip arrays to identify the changes in the DNA methylation level between the two groups. Then, deep bisulphite sequencing was used to validate five CpGs on 78 samples. The results from the Infinium 450K found that only 11 CpGs showed a significant difference in DNA methylation between the case and the control groups. Five CpGs of the eleven (cg00648582, cg0932376, cg19169023, cg23841288 and cg27391564) underwent deep bisulphite sequencing where cg00648582, related to the PGAM5 gene, and the cg23841288 CpGs, related to the PTPRN2 gene amplicons, showed a significant increase in their DNA methylation level in more than one CpG in the case group. In contrast, a significant decrease was found at cg19169023 and at its various neighbouring CpGs in the TYRO3 gene‐related amplicons. Furthermore, this study demonstrated a significant correlation between the variation in sperm DNA methylation level and standard sperm parameters in the case group.     

Influence of extended incubation time on Human sperm chromatin condensation,sperm DNA strand breaks and their effect on fertilisation rate

The purpose of this study was to determine influence of extended incubation time on sperm chromatin condensation and DNA strand breaks and their effect on fertilisation rate. Forty couples undergoing ICSI therapy were included. Semen was prepared by PureSperm gradient centrifugation and divided into two parts. The first part (G1) was used immediately for ICSI, whereas the second part (G2) was kept in the incubator at 37°C, 5% and 90% Humidity for 5 hr, and thereafter, the capacitated spermatozoa were used for ICSI. The TUNEL test and chromomycin CMA3 were used to evaluate the DNA strand breaks and chromatin condensation respectively. The percentage of condensed chromatin was 73.92 ± 12.70 in the group 1 and 81.13 ± 10.31% in group 2 (p = .001). However, the double‐strand breaks were 11.15 ± 8.67% in G.1 and 16.30 ± 11.12% in G.2. (p = .001). Fertilisation rate in the (Group 1) was 62.45% and 69.17% in (Group 2). There was a positive correlation between condensed chromatin and fertilisation rate (r = 0.846, p = .001) and a negative correlation with DNA double‐strand breaks (r = ?0.802; p = .001). In conclusion, the prolonged sperm incubation (5 hr) leads to a higher chromatin condensation and to a significantly increased number of DNA strands double breaks with no influence on fertilisation rates.     

大鼠膜联蛋白5的重组表达及其对人精子体外运动功能的影响

目的:先前研究发现促性腺激素释放激素(GnRH)激动剂对大鼠睾丸间质细胞中膜联蛋白5(annex-in5)的翻译和转录水平都有一定的影响,推测annexin5可能是睾酮分泌调节中的一个信号分子。为研究annexin5在雄性生殖调节中的作用,本研究拟获得具有活性的大鼠annexin5重组蛋白,并观察其对人精子运动功能的影响。方法:化学合成法合成大鼠annexin5的编码基因序列,将该基因插入组氨酸标签(HIS)融合表达载体pET28a中,在T7启动子控制下,通过异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达HIS融合蛋白;以亲和层析法纯化表达融合蛋白并用活化部分凝血激酶时间(APTT)交叉试验法验证该蛋白的抗凝血活性。15例供精者的精液标本一式2份,1份加重组蛋白annexin5(终浓度为10-8mol/L),1份做对照,分别处理20和60min,用计算机辅助精液分析系统(CASA)分析精子活力、活动率;处理20min后,做精子爬高试验。结果:合成的目的基因产物长度为974bp,插入序列与annexin5的序列完全一致。在IPTG诱导下,BL21(DE3)重组菌高效表达出相对分子质量为36000的融合蛋白,经纯化获得95%纯度的融合蛋白,纯化蛋白具有很强的抗凝血活性。Annexin5处理20min后精子活动率提高了21%(P<0.05),活力提高了40%(P<0.01),而处理60min后精子活力、活动率与对照组比较均无显著性差异;精子爬高高度与对照组比较具有极显著差异[(37.84±6.35)mmvs(49.5±12.27)mm,P<0.01]。结论:成功构建了大鼠annexin5重组表达载体并在大肠埃希菌中高效表达annexin5蛋白,且该蛋白体外处理精子20min时提高精子的运动能力。     

Cryopreservation of human sperm: efficacy and use of a new nitrogen-free controlled rate freezer versus liquid nitrogen vapour freezing

Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory.     

Human sperm DNA integrity in normal and abnormal semen samples and its correlation with sperm characteristics

Reports indicate an increase in the incidence of DNA fragmentation in male factor infertility and its role in the outcome of assisted reproductive techniques (ART). However, reports are conflicting between the relationships of sperm DNA integrity with conventional semen parameters. We examined the relationship between conventional sperm parameters and DNA integrity using acridine orange (AO) test. The study included 373 patients and 28 fertile volunteers. DNA normality was compared with semen parameters between the patient and donor populations. Significant correlations were noted between DNA normality and sperm concentration ( r  = 0.18, P  = 0.000), motility ( r =  0.21, P  = 0.0001), rapid motility (0.19, P  = 0.000), normal morphology by World Health Organization ( r =  0.15, P  = 0.019) and head defects ( r =  −0.15, P  = 0.023). A significant difference was noted in AO levels between donors and patients with asthenozoospermia ( P  = 0.002) and oligoasthenozoospermia ( P  = 0.001). A significant difference in DNA integrity was noted in samples having <30% and >30% normal morphology. A wide range of % DNA normality was observed in the patient group. Sperm assessment for DNA status using AO is reliable and shows good correlation with sperm count, motility and morphology. Assessment of sperm DNA status with AO staining may be helpful prior to ART.     

Seminal parameters of chronic male genital inflammation are associated with disturbed sperm DNA integrity

Definition of chronic male genital tract inflammation and its impact on male infertility is still a matter of debate. In particular, DNA integrity has been reported to be disturbed in subfertile men. Thus, the aim of this study was to investigate an association of DNA integrity to altered standard semen parameters as well as inflammatory parameters such as peroxidase‐positive cells, macrophages and seminal interleukin‐6 concentration. Macrophages were detected by CD18/HLA‐Dr staining, and DNA integrity was analysed by acridine orange staining using flow cytometry. Interleukin‐6 was detected by ELISA. Normal DNA integrity showed a significant correlation to sperm number and progressive motility. Moreover, a significant inverse correlation of DNA integrity to Interleukin‐6 and macrophages could be demonstrated. Further on, seminal interleukin‐6 also significantly correlated to macrophages. No association has been observed between the number of peroxidase‐positive cells and normal DNA integrity. As disturbed DNA integrity has been reported to negatively influence spermatozoon–egg interaction and even fertilisation rates following ICSI, and as early miscarriages have been associated with sperm DNA damage, it should be screened very carefully for male genital tract inflammations in couples undergoing infertility treatment. Measuring Interleukin‐6 seems superior to assessment of the number of leucocytes alone and additional assessment of DNA integrity into the diagnostic work‐up should be considered.     

The presence of human papillomavirus in semen does not affect the integrity of sperm DNA

It remains unknown whether human papillomaviruses (HPVs) in semen affect sperm DNA integrity. We investigated whether the presence of these viruses in semen was associated with an elevated sperm DNA fragmentation index. Semen samples of 22 normozoospermic patients undergoing infertility treatment, nine fertile donors and seven fertile men with a risk of HPV infection (genital warts or condylomas) were included in the study. The samples were examined by an INNO‐LiPA test PCR‐based reverse hybridisation array that identifies 28 types of HPVs as simple or multiple infections. Sperm DNA integrity was determined by sperm chromatin dispersion assay (SCD). Our preliminary findings demonstrate an increase in HPV infection in infertile men with respect to fertile men. However, the sperm DNA fragmentation index was not increased in semen containing these viruses.     

Stimulation of sperm motility by kinins in fresh and 24 hours aged human ejaculates

Kallikrein, a kinin-releasing proteinase, significantly stimulates human sperm motility in fresh ejaculates with primarely reduced sperm motility (asthenozoospermia) as well as in 24 hours stored ejaculates with reduced sperm motility resulting from post-ejaculatory aging processes. adiition of a kininogen source (serum) further significantly improved sperm motility of kallikrein-stimulated ejaculates. In contrast to the 24 hours lasting stimulation of kallikrein, enhancement of sperm motility by kinins is limited to 2 hours after addition of a single dose of kallidin (1 ng/ml).     

影响人类精子耐冻性的蛋白质分析

目的:利用比较蛋白质组学技术研究影响人类精子耐冻性的蛋白质。方法:将10例供精志愿者的31份精液标本依据冷冻复苏率的不同分为高冷冻复苏率组和低冷冻复苏率组,应用二维电泳技术和质谱分析鉴定技术寻找两组标本的精子蛋白和精浆蛋白在表达量上的差异。结果:在精子蛋白和精浆蛋白中共发现了22个差异表达蛋白。其中精浆蛋白12个,分别为细胞角蛋白1、神经元导向因子3、Rho2样蛋白、Krueppel样因子10、FERM域蛋白8、GMP合成酶、鸟嘌呤结合蛋白G、溶质载体家族12、ATP结合盒亚家族D1、6-磷酸果糖-2-激酶、肌钙蛋白C、肌球蛋白轻链;精子蛋白9个,分别为卷曲螺旋结构域蛋白113、RAF原癌基因丝氨酸/苏氨酸蛋白激酶、Ras相关蛋白Rab-31、α微管蛋白3C/D、锌指蛋白879、胆固醇26-羟化酶、DNA修复和重组蛋白、细胞角蛋白73、钙磷蛋白;精浆和精子共有蛋白1个,为细胞周期依赖性蛋白激酶1。文献调研发现这些差异蛋白主要与精子的能量代谢、成熟、运动、DNA修复等有关。结论:鉴定了多种可能影响人类精子耐冻性的蛋白质,为以后从分子水平上揭示精子冷冻复苏原理提供了依据。     

膜联蛋白5对人精子膜及DNA完整性的影响

目的:研究膜联蛋白5(Annexin5)对人精子细胞膜及DNA完整性的影响。方法:①精子膜完整性测定:按精子浓度>20×106/ml;活率>60%选取标准收集精液标本53份,分为3组,实验组为47.5μl精液中加入2.5μl10-6mol/L的Annexin5;阴性对照组为47.5μl精液标本中加入2.5μl1mol/L的Tris-HCl(pH8.0,25℃);空白对照组为47.5μl精液标本中加入2.5μl0.01mol/L的PBS(pH7.4),作用20min后,通过精子低渗肿胀试验(HOS)检测精子膜的完整性。②DNA完整性测定:同方法①,3组实验标本作用20min后,每份标本加入2.5μl0.02mol/L的H2O2,作用60min,通过吖啶橙(AO)荧光染色检测精子核DNA的完整性。结果:An-nexin5处理20min后低渗肿胀精子百分率与空白对照组及阴性对照组比较均具有极显著差异[(66.17±12.02)%vs(58.13±13.08)%,P<0.01;(66.17±12.02)%vs(59.94±11.91)%,P<0.01];空白对照组与阴性对照组比较无显著性差异。加入H2O2后,Annexin5组的DNA碎片指数与空白对照组及阴性对照组比较均具有极显著差异[(6.39±1.07)%vs(11.16±1.16)%,P<0.01;(6.39±1.07)%vs(10.86±1.05)%,P<0.01],空白对照组与阴性对照组比较无显著性差异。结论:Annexin5蛋白能在体外提高低渗肿胀精子百分率,对精子膜的完整性具有保护作用,同时对HO作用引起的精子核DNA破坏起保护作用。     

Cryoprotective effect of l‐carnitine on motility,vitality and DNA oxidation of human spermatozoa

Successful cryopreservation for human spermatozoa markedly influences the reproductive outcomes of assisted reproductive technologies. But in spite of its usefulness, cryopreservation significantly decreases sperm quality. l ‐carnitine has been found to improve the quality of spermatozoa in selected cases with male infertility. Here, we examined the efficacy of l ‐carnitine in improving sperm motility and vitality and reducing sperm DNA oxidation during cryopreservation. Semen samples from infertile patients (n = 22) were collected and analysed. Cryopreservation medium supplemented with l ‐carnitine was mixed with the semen at a ratio of 1 : 1 (v/v). The final l ‐carnitine concentration in each cryovial was 0.5 mg ml^?1 per 5 × 10⁶ cell ml^?1. Controls were cryopreserved without addition of l ‐carnitine. After 24 h of cryopreservation, thawed sperm samples were analysed for motility, vitality and DNA oxidation. Sperm vitality was assessed by the eosin–nigrosin test, while sperm DNA oxidation was measured by flow cytometry. Addition of l ‐carnitine significantly improved sperm motility and vitality (P < 0.05) compared with the control. The flow cytometry experiment showed no statistical difference (P > 0.05) in the levels of DNA oxidation between samples and controls. In conclusion, l ‐carnitine improves human sperm motility and vitality, but has no effect on sperm DNA oxidation after cryopreservation.     

Assessment of sperm quality, DNA integrity and cryopreservation protocols in men diagnosed with testicular and systemic malignancies

Men diagnosed with malignancy are often referred for semen banking to preserve their fertility prior to cancer treatment. The chances of cancer patients for achieving future fecundity will be determined by the sperm quality including the integrity of the genomic material in the frozen samples. The objectives of this study were to compare the sperm quality and DNA integrity in men diagnosed with testicular and systemic malignancies before receiving treatment and to identify the optimum cryopreservation protocol for their samples including a remote semen collection option. In comparison with fertile donors, patients with testicular malignancies had significantly lower sperm concentration, while both testicular and systemic malignancy patients had significantly lower sperm motility and cryosurvival rates. In addition, the SCSA defined DNA fragmentation index was significantly higher in patients with testicular and systemic malignancies compared with fertile donors. It was noted that the extent of deterioration in sperm quality and DNA integrity seen in cancer patients did not reach the previously defined statistical threshold for impaired fertility. Freezing spermatozoa with the seminal plasma offers the highest protection against cryo-injury. Nevertheless, remote semen collection can still be used as it yields adequate results.