The relationship of sperm DNA fragmentation index with the outcomes of in-vitro fertilisation-embryo transfer and intracytoplasmic sperm injection

This study was conducted to investigate the relationship between sperm DNA fragmentation index (DFI) and the outcomes of in-vitro fertilisation-embryo transfer (IVF-ET) and intracytoplasmic sperm injection (ICSI). Sperm DFI in 116 patients for IVF and 63 patients for ICSI were detected with acridine orange test (AOT), and all the cases were divided into DFI ≤ 30% group and DFI > 30% group according to the DFI value; then, the relationship of DFI with the outcomes of IVF/ICSI were analysed. Both in IVF and ICSI cycles, good embryo rate and spontaneous abortion rate in DFI > 30% group were significantly different from that in DFI ≤ 30% group, meanwhile, the fertilisation rate and cleavage rate were similar in two groups. In ICSI cycles, there was a significantly negative correlation between the DFI value and the rates of embryo implantation and pregnancy; the couples with DFI > 30% had significantly lower embryo implantation rate and pregnancy rate than the ones with DFI ≤ 30%. The receiver operating characteristics curve analysis demonstrated that the DFI value were statistically significant predictors of pregnancy. It is concluded that DNA-damaged sperm could have a potential adverse effect on embryo quality and progression of pregnancy as well as the outcomes of ICSI.     

Sperm DNA fragmentation index,as measured by sperm chromatin dispersion,might not predict assisted reproductive outcome

Objective

Routine semen parameters have limited clinical diagnostic value for predicting male infertility. The aim of this study was to investigate the association between sperm DNA fragmentation index (DFI) and semen quality, and between DFI and clinical pregnancy rate of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI).

Female age affects the utility of sperm DNA fragmentation in predicting IVF and ICSI outcomes

Research questionThe aim of this study was to investigate how female age affects the predictive effect of sperm DNA fragmentation index (DFI) on clinical outcomes with assisted reproductive technology.DesignA total of 2371 patients, comprising 2115 men with a normal DFI (≤30), 256 men with a high DFI (>30) and women of different ages, were recruited and investigated. All patients had normal chromosome karyotypes and were undergoing their first fresh IVF or intracytoplasmic sperm injection (ICSI) cycles. Clinical outcomes were analysed according to the two DFI groups and female age ≤30 and >30 years. Binary logistic regression analysis was performed to identify factors associated with clinical outcome.ResultsThe proportion of couples with at least one good-quality embryo in the DFI ≤30 group was higher than that in the DFI >30 group. When female age exceeded 30 years, clinical pregnancy rate and the proportion of couples with good-quality embryos in the DFI >30 group were lower compared with DFI ≤30; however, there were no differences in outcomes for female age ≤30 years according to DFI. When DFI >30, the cut-off value of female age was 30.5 for detecting clinical pregnancy; the sensitivity was 62.0%, and the specificity was 63.6%. Clinical pregnancy rate and proportion of couples with good-quality embryos were lower in the DFI >30 versus DFI ≤30 group with a female age above 30 years for IVF but not for ICSI.ConclusionFemale age has a negative effect and should be considered in predicting the effects of sperm DNA fragmentation on pregnancy outcomes.     

精子DNA碎片与体外受精结局的关系

目的:探讨精子DNA碎片与体外受精(in vitro fertilization,IVF)结局的关系。方法:采用染色质扩散实验(sperm chromatin dispersion,SCD)对242例接受IVF的男方进行精子DNA碎片率(DNAfragmentation index,DFI)检测,按照WHO标准进行精液常规分析,将精子DFI、精液常规参数和IVF受精率、卵裂率、可移植胚胎率、优质胚胎率进行Spearman相关分析,将精子DFI、精液常规参数对生化妊娠、临床妊娠的影响进行Logistic回归分析。结果:精子DFI与精子前向活动率呈负相关(r=-0.355,P<0.001);密度梯度法处理前、后精子DFI均与IVF受精率呈负相关(r=-0.223,P<0.001)(r=-0.136,P<0.05);精子DFI、精液常规参数与卵裂率、可移植胚胎率、优质胚胎率无相关性;精子DFI与生化妊娠、临床妊娠结局无相关性。结论:精子DFI影响精子活力与IVF受精率,精子DFI检测对预测IVF受精率有一定的临床意义。     

Clinical significance of sperm DNA damage threshold value in the assessment of male infertility

Sperm DNA integrity is a prerequisite for normal spermatozoal function. The aim of the study was to evaluate the role of sperm chromatin damage, its cut-off level and its effect on sperm parameters in men with idiopathic infertility by analyzing 100 idiopathic infertile men and 50 fertile controls. Semen samples were analyzed as per WHO 1999 guidelines and sperm chromatin structure assay (SCSA) was applied to measure DNA fragmentation index (DFI) in sperm. The mean DFI of infertile men (35.75) was significantly (P < .0001) higher as compared to controls (26.22). The threshold level of 30.28% was obtained as cut-off value to discriminate infertile men from fertile controls. Sperm count, forward motility, and normal morphology found to be negatively associated with DFI in overall study subjects. Infertile men with severe oligozoospermia had higher mean DFI (40.01 ± 11.31) than infertile men with oligozoospermia (35.11 ± 10.05) and normal sperm count (33.99 ± 9.96). Moreover 64% of infertile men have DFI > 30 against 6% of fertile controls (P < .0001). Higher sperm DNA fragmentation may be the underlying cause for poor semen quality in idiopathic infertile men and the threshold value of 30.28% is a clear discriminator to distinguish infertile men from fertile men of Indian population. Thus, DFI is a good prognostic marker as cases with higher sperm DFI may have poor success rate even after assisted conception and may experience recurrent pregnancy loss (RPL) and should be counseled accordingly.     

Impact of sperm DNA fragmentation on ART outcome

We have used the sperm chromatin structure assay (SCSA) test in order to determine if correlations can be found between sperm DNA fragmentation and spermogram parameters. Only necrospermia and DNA fragmentation index are strongly correlated (P<0.0001). Neither fertilization rates for ICSI and IVF, nor blastocyst formation rates are impaired by a high DFI. However when the critical DFI>30% is reached, the chances of having ongoing pregnancies after blastocyst transfer are reduced by three. Treatments with antioxidants are of limited efficacy even though we obtained 2 deliveries after DFI treatments with such treatments. New strategies in order to improve the pregnancy rates for these peculiar cases of reduced fertility are discussed.     

中药生精散对精子DNA完整性的影响及其在IVF-ET治疗中的临床疗效

目的:观察中药生精散对不育患者精子DNA完整性的影响及其在IVF-ET治疗中的临床疗效。方法:因少、弱精子症行IVF/ICSI-ET治疗的患者49例,随机分为中药组(22例,服用生精散治疗)和对照组(27例,未经生精散治疗)。观察比较中药组和对照组精液常规分析(精子密度、活率、活力)、DNA碎片指数(DNA fragmentation index,DFI)、受精率、卵裂率、种植率及临床妊娠率。结果:中药治疗后精子活率及活力显著提高(P<0.05),且中药组DFI值较治疗前及对照组明显下降(P<0.05)。中药组胚胎种植率(31.11%)及临床妊娠率(45.45%)明显高于对照组(13.73%和18.51%)(P<0.05)。结论:中药生精散可通过降低不育患者精子DNA损伤程度,改善精子质量,提高IVF-ET的治疗效果。     

Sperm DNA fragmentation index and pregnancy outcome after IVF or ICSI: a meta-analysis

PurposeThe purpose of this study was to carry out a meta-analysis for a comprehensive understanding and estimation of the association between sperm DNA Fragmentation Index (DFI) and pregnancy outcome after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatment.MethodsStudies concerning the link of DFI with pregnancy outcome were included after literature search of database PUBMED, EMBASE, MEDLINE. Related information was extracted from the eligible studies by two independent authors and a meta-analysis was conducted by using STATA 12.0 software. Pregnancy outcomes consisted of biochemical pregnancy (BP), clinical pregnancy (CP) and pregnancy loss (PL). The studies included for meta-analysis were divided into three groups according to the DFI threshold value (DFI >27 %, 15–27 %, ≤15 % group). The odds ratio (OR ) and their 95 % confidence intervals (95 % CIs) were used to evaluate the association between DFI and pregnancy outcome.ResultsTwenty articles were included in our meta-analysis. The results indicated that infertile couples were more likely to get pregnant if DFI was less than threshold value (For threshold value > 27 % and 15–27 % group, combined overall OR (95 % CI) = 1.437 (1.186–1.742), 1.639 (1.093–2.459) respectively). However, when stratified by DFI detection methods, using sperm chromatin structure assay (SCSA) as the DFI test method, the results indicated a similar CP rate between groups with a high DFI or a lower DFI value (SCSA, For threshold value >27 % and 15–27 % group, combined overall OR (95 % CI) = 1.242(0.978–1.577), 1.480(0.921–2.377) respectively). The meta-analysis based on BP (overall OR (95 % CI) = 0.952 (0.697–1.302)) and PL((For DFI >27 %, 15–27 %, ≤15 % group, OR (95 % CI) = 0.786 (0.491–1.258), 1.509 (0.655–3.476), (0.538 (0.264–1.097) respectively) outcome yielded nonsignificant results.ConclusionsThe predication value of DFI for IVF or ICSI outcome is not confirmed in our meta-analysis. Further better designed studies with larger subjects involved are needed to better address this issue.     

Relationship between the outcomes of assisted reproductive techniques and sperm DNA fragmentation as measured by the sperm chromatin structure assay

OBJECTIVE: To investigate how moderate and/or high levels of DNA fragmentation (DFI), as measured by the sperm chromatin structure assay (SCSA), affect either IVF or IVF with intracytoplasmic sperm injection (ICSI) fertilization, cleavage, blastulation, implantation, and pregnancy. DESIGN: Retrospective clinical study. SETTING: Academic human reproduction laboratory. PATIENT(S): Eighty-nine couples undergoing IVF with conventional fertilization or ICSI. INTERVENTION(S): Sperm chromatin structure assay testing (SCSA) of semen aliquot taken from ejaculate used for assisted reproductive technology (ART). MAIN OUTCOME MEASURE(S): Related DFI to conventional semen parameters and cycle-specific outcomes after ART. RESULT(S): No patients achieved clinical pregnancy if SCSA values exceeded the DFI (27%, P<.01), moderate DFI (15%, P<.01), or high DFI (15%, P<.05) thresholds. Dividing the DFI sperm population into moderate-fragmentation and high-fragmentation categories did not improve the prognostic value of the SCSA. No coefficient of determination (r(2)) between SCSA parameters and conventional parameters exceeded 0.29. CONCLUSION(S): Sperm chromatin structure assay identified thresholds for negative pregnancy outcome after ART not identified using conventional semen parameters. This is the first study analyzing the clinical value of sperm DFI to [1] include a large number of ART patients (n = 89), [2] perform SCSA analysis on a semen aliquot from the ejaculate used for ART, and [3] examine how the extent (moderate and high DFI) of DFI influenced ART outcomes.     

Predictive value of DNA integrity analysis in idiopathic recurrent pregnancy loss following spontaneous conception

Purpose

Standard semen parameters are poor predictors of fertility potential. To date, apart from, paternal karyotyping sperm factors are not evaluated in recurrent pregnancy loss (RPL), only recent studies have emphasized the role of sperm factors in early embryonic development as sperm transcribes genes critical for early embryonic development. Sperm DNA integrity is useful diagnostic and prognostic marker and has clinical implications in idiopathic recurrent pregnancy loss (iRPL) following spontaneous conception. The aim of this study was to assess DNA integrity in cases experiencing iRPL following spontaneous conception.

Methods

Semen samples from 45 patients and 20 controls were analyzed as per WHO 1999 guidelines and sperm chromatin structure assay (SCSA) was used to measure DNA fragmentation index (DFI).

Results

By applying receiver operating curve (ROC) analysis, sperm DFI of approximately 26 % was found in male partner of couples experiencing iRPL.

Conclusions

Our data indicate that sperm from men with a history of iRPL have a higher percentage of DNA damage as compared to control group, and this can explain pregnancy loss in these patients. Men with higher DFI are infertile whereas men with lower DFI (26 %) are able to conceive but experience recurrent pregnancy loss. Thus it is important to evaluate sperm DFI in couples experiencing iRPL to understand exact aetiology of RPL and determine prognosis and management.     

Assessment of density gradient centrifugation (DGC) and sperm chromatin dispersion (SCD) measurements in couples with male factor infertility undergoing ICSI

Purpose

To investigate how effectively density gradient centrifugation (DGC) improves sperm nuclear integrity and to determine whether the sperm chromatin dispersion (SCD) test of sperm nuclear integrity in native or DGC-treated semen can predict the outcome of assisted reproductive technology (ART) in couples undergoing intracytoplasmic sperm injection (ICSI).

Methods

The DNA integrity of spermatozoa from 63 male factor infertility patients undergoing ICSI was analyzed by the SCD test before and after DGC. The predictive value of the sperm DNA fragmentation index (DFI) for ART outcomes was assessed in a cohort of 45 patients who were undergoing fresh embryo transfer. For the analysis, they were divided into pregnant and non-pregnant groups and, independently, into high sperm DFI (DFI > 30 %) and low sperm DFI (DFI ≤ 30 %) groups. Both raw and DGC semen parameters were examined.

Results

In the asthenospermia and oligozoospermia groups, DGC decreased the sperm DFI from 31.5 ± 19.7 and 28.5 ± 10.3 to 19.2 ± 18.3 and 16.0 ± 12.8, respectively (P < 0.01). DGC decreased the sperm DFI in the severe oligozoospermia group from 41.4 ± 19.0 to 36.3 ± 20.6 (P > 0.01). The pregnant and non-pregnant groups did not differ in their fertilization rate and sperm DFI in native or DGC semen (P > 0.05). There was also no significant difference between the high sperm DFI (DFI > 30 %) and low sperm DFI (DFI ≤ 30 %) groups with regard to fertilization rate, implantation rate, and clinical pregnancy rate for both native and DGC semen (P > 0.05). The patients undergoing ICSI with a high sperm DFI had a higher pregnancy loss rate (defined as spontaneous miscarriage or biochemical pregnancy) compared with patients with a low sperm DFI in both the native and DGC semen groups.

Conclusions

DGC highly significantly reduces sperm DNA fragmentation in the semen of ICSI patients, with the exception of those with severe oligozoospermia. The results of the SCD test of sperm DNA fragmentation in native or DGC semen do not correlate with the fertilization rate, implantation rate, or clinical pregnancy rate in patients undergoing ICSI.     

DNA fragmentation of spermatozoa and assisted reproduction technology

Despite the ever-increasing knowledge of the fertilization process, there is still a need for better understanding of the causes of sperm DNA fragmentation and its impact on fertilization and pregnancy. For this reason, human sperm DNA fragmentation was investigated by means of the terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling (TUNEL) assay and the production of reactive oxygen species (ROS) in the ejaculate and in the spermatozoa themselves. These data were correlated with fertilization and pregnancy data from IVF and intracytoplasmic sperm injection (ICSI) patients. Sperm DNA fragmentation did not correlate with fertilization rate, but there was a significantly reduced pregnancy rate in IVF patients inseminated with TUNEL-positive spermatozoa. ICSI patients exhibited the same tendency. This implies that spermatozoa with damaged DNA are able to fertilize an oocyte, but at the time the paternal genome is switched on, further development stops. The determination of ROS in the ejaculate and the percentage of ROS-producing spermatozoa revealed markedly stronger correlations between sperm functions (i.e. motility) and the percentage of ROS-producing spermatozoa. The influence of seminal leukocytes, known to produce large amounts of oxidants, on sperm DNA fragmentation should not be neglected.     

Data analysis of two in vivo fertility studies using Sperm Chromatin Structure Assay-derived DNA fragmentation index vs. pregnancy outcome

The 2006 American Society for Reproductive Medicine Compendium of Practice Report found no significant effects of elevated sperm DNA fragmentation by using a 30% DNA fragmentation index (DFI) threshold for natural fertilization and SCSA data (odds ratio, 1.07; 95% confidence interval, 0.39-2.93). In contrast, it is shown in this article that these two in vivo studies showed significant odds ratios of 6.54 (95% confidence interval, 1.71, 24.91) and 7.58 (95% confidence interval, 2.54, 22.67), which resulted in the conclusion that the pregnancy (PG) rates are statistically significantly higher for the group with DFI below the thresholds of 30% and 40%, respectively. In addition, all the chi(2) statistics used to test the null hypotheses of no association between the pregnancy status and DFI for natural (normal) fertilization confirmed this conclusion that the probability of pregnancy for the group with <30% or <40% DFI was statistically significantly higher than that for the group with >30% or >40% DFI.     

Sperm DNA fragmentation and male fertility: a retrospective study of 5114 men attending a reproductive center

PurposeThe sperm DNA fragmentation index (DFI) was quantitatively measured and its relationship with age, semen quality, and infertility conditions was investigated.MethodsSemen routine test and sperm DFI were performed in 2760 infertile male and 2354 male whose spouse experienced at least one unexplained miscarriage to analyze the correlation between sperm DNA damage, semen routine parameters, and age.ResultsSperm DFI was significantly lower from patients whose wife experienced unexplained miscarriage compared to infertility males (p = 0.000). An inverse correlation between sperm DFI and sperm progressive motility was observed (r_s = − 0.465, p = 0.000) and sperm DFI was positively correlated with age (r_s = 0.255, p = 0.000). However, the correlation between sperm DFI and sperm concentration, semen volume, total sperm count, and motile sperm count were not proved.ConclusionsSperm DFI is an important indicator for evaluating the quality of semen. Sperm DNA integrity testing is preferentially recommended to those who have decreased sperm progressive motility, especially older men. An integrative analysis of sperm DFI, sperm progressive motility, age, and infertility conditions can provide a more comprehensive assessment of male fertility.     

Relationship between sperm DNA damage, induced acrosome reaction and viability in ICSI patients

The DNA damage in human spermatozoa is a relevant predictor of prognosis in male infertility, whereby increased sperm DNA damage impairs the outcomes of artificial reproduction. Theoretically, DNA damage should alter the special cellular functions of human spermatozoa, and lead to diminished acrosome reaction with reduced fertilization rates. Nevertheless, intracytoplasmic sperm injection (ICSI) has been reported to alleviate such negative outcomes due to DNA damage. This study investigated the relationship between DNA fragmentation and acrosome reaction as well as viability in ICSI patients. The study enrolled 42 men undergoing ICSI due to poor sperm parameters. The DNA fragmentation indexes (DFI) were 4-10% in 38% of the cases, and > or = 10% in 19% of the cases. The results of both acrosome reaction and viability assays showed negative correlations with DFI values in all cases and especially in cases with fertilization rates <60% (P < 0.05). However, such correlations were not found in cases with fertilization rates >60%. There were no live deliveries in patients with high DFI levels (>10%). In conclusion, negative correlations were identified between increased DNA damage, and acrosome reaction and/or viability of human spermatozoa, especially in cases with reduced fertilization rates.     

Clinical implications of sperm DNA damage

Traditionally, the diagnosis of male infertility has relied upon microscopic assessment and biochemical assays to determine human semen quality. These tests are essential to provide the fundamental information on which clinicians base their initial diagnosis. However, none of these parameters addresses sperm function and their clinical value in predicting fertility is questionable. The advent of intracytoplasmic sperm injection (ICSI) has further reduced the significance and perceived need for sperm quality tests since ICSI requires only one sperm for the procedure to be successful. Even the conventional measures of sperm quality in terms of normal morphology or motility are not necessary for successful ICSI. Funding of andrological research has been neglected and improvement in assisted reproductive technology (ART) success has suffered as a consequence. Testing of sperm DNA damage shows much promise both as a diagnostic test for male infertility and a prognostic test for ART outcomes. It has been shown to be closely associated with numerous fertility outcomes including negative relationships with fertilization, embryo quality, implantation and positive relationships with miscarriage and childhood diseases. Here we report the relationships between in?vitro fertilisation, ICSI pregnancy rates and sperm DNA damage, using the Comet assay to measure DNA fragmentation and also a novel test to measure modified bases, as a indication of oxidative DNA injury.     

Negative impact of cigarette smoking on male fertility: from spermatozoa to the offspring

Cigarette smoking has negative effects on male fertility. Recent studies showed an active transfer of several components of cigarettes through the blood-testis barrier. The presence of these components in the seminal plasma may induce a degradation of sperm parameters and nuclear quality of spermatozoa, and compromise the chances of pregnancy. Moreover, smoking may have a negative impact on the smokers'offspring: poor quality embryos, development of childhood cancers. Oxidative stress-induced DNA damage seems to be one of the major causes of sperm quality alteration. Several methods are now available to analyze the degree of DNA fragmentation. In order to optimize the success rate of assisted reproduction technologies, the deleterious effects of smoking on male fertility and the necessity of cessation have to be explained in detail to these patients.     

Correlation between the DNA fragmentation index (DFI) and sperm morphology of infertile patients

Purpose

To evaluate the correlation between the DNA Fragmentation Index (DFI) and sperm morphology in patients undergoing ICSI, as a predictive parameter in reproductive outcomes.

Methods

A retrospective study was conducted on 125 infertile patients enrolled in a fertility clinic. Seminal characteristics were measured following the WHO guidelines (2010) for the examination of the seminal fluid. After collecting motile sperm population by pellet swim up, DFI was calculated and simultaneously associated with sperm morphology using in situ TUNEL assay and an image analyzer software in at least 250 spermatozoa for each patient.

Results

All subjects were divided into two groups according to a cutoff established, by choice, of the sperm DFI (15%): group A (< 15%) consisting of 65 patients and group B (≥ 15%) of 60 patients. Data were analyzed using non-parametric statistical methods. The results demonstrate that there is no statistical difference between the two groups in seminal characteristics. The collective data show a high significant correlation, suggesting that spermatozoa with abnormal morphology are the best candidates to contain DNA damage (p < 0.001). Also, when group A is compared with group B, an increased percentage of morphologically normal spermatozoa with fragmented DNA was observed in patients, with DFI values ≥ 15% (p < 0.001).

Conclusion

These results are aimed at providing an exact value of DFI in morphologically normal spermatozoa, which will be helpful to the embryologist in evaluating the risk of transferring, during the ICSI procedure, a spermatozoon whit normal morphology but fragmented DNA.

     

Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles

OBJECTIVE: To determine the relationship between sperm chromatin structure assay (SCSA) parameters (DNA fragmentation index [DFI] and high DNA stainability [HDS]), and conventional IVF and IVF/intracytoplasmic sperm injection (ICSI) outcomes. DESIGN: Retrospective review and prospective study. SETTING: Private IVF clinic. PATIENT(S): Two hundred forty-nine couples undergoing first IVF and/or ICSI cycle. INTERVENTION(S): IVF, ICSI, blastocyst culture. MAIN OUTCOME MEASURE(S): DFI, HDS, conventional semen parameters, IVF, ICSI. RESULT(S): IVF and ICSI fertilization rates were not statistically different between high- and low-DFI groups. More men with > or =15% HDS had lower (<25% and <50%) IVF fertilization rates. High DNA stainability was not related to ICSI fertilization rates. High DNA stainability did not affect blastocyst rates or pregnancy outcomes. Men with > or =30% DFI were at risk for low blastocyst rates (<30%) and no ongoing pregnancies. Men with > or =30% DFI had more male factors. World Health Organization thresholds were not predictive of ongoing pregnancy. CONCLUSION(S): The relationship between HDS and poor IVF fertilization rates provides preliminary evidence that ICSI may be indicated in men with > or =15% HDS. Men with high levels of DNA fragmentation (> or =30% DFI) were at greater risk for low blastocyst rates and failure to initiate an ongoing pregnancy. The SCSA provides valuable prognostic information to physicians counseling couples before IVF and/or ICSI cycles.     

Efficacy of swim-up versus density gradient centrifugation in improving sperm deformity rate and DNA fragmentation index in semen samples from teratozoospermic patients

Purpose

To compare the efficacy of swim-up and DGC in improving sperm deformity and DNA fragmentation and to determine which method is better in teratozoospermic patients requiring artificial reproduction.

Methods

The present study compared the effects of swim-up and density gradient centrifugation (DGC), the two most commonly used semen preparation methods, on sperm deformity rate and DNA fragmentation index (DFI) in semen samples from teratozoospermic patients.

Results

The results demonstrated that both swim-up and DGC yielded a significantly lower sperm deformity rate and DFI in comparison to unprocessed whole semen, with DGC having more favorable results. Sperm deformity rate in unprocessed whole semen samples was significantly lower in the 20–29 age group than in the 40-49 age group, but no significant difference was observed in DFI between different age groups. There was no significant correlation between sperm deformity rate and DFI.

Conclusions

Our findings suggest that enrichment of sperm with normal morphology and intact DNA in teratozoospermic patients could be achieved by both DGC and swim-up procedures, and that DGC is a better method.