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1.
The study was aimed to investigate the association between the degree of oligozoospermia and sperm chromosome aneuploidy frequencies in male infertility and to determine whether chromosomal profiles of sperm nuclei would be used for a supportive test before additive reproduction technics. The meiotic segregation profiles of chromosomes X, Y, 13, 18 and 21 were compared by fluorescent in‐situ hybridisation (FISH) on the spermatozoa of 30 normally karyotyped oligozoospermic (10 mild, 11 moderate, nine severe) cases without Y‐microdeletions, and 10 normozoospermic cases. The results showed significantly higher frequencies of chromosomes 13, 18, 21 disomies (P < 0.001) in the group of patients with moderate and severe oligozoospermia compared with the disomy frequencies of normozoospermic group. The statistically significant differences were also determined in disomy frequencies of sex chromosomes (XY, XX and YY) in between oligozoospermic and normozoospermic groups (P < 0.001, P < 0.001, P < 0.040, respectively). Because oligozoospermic patients are the ones consulted the most for assisted reproductive techniques, identification of sperm aneuploidy rates in men could be considered as an appropriate supportive test before the reproductive implementations. Furthermore, the patients should be counselled with respect to genetic screening results for the potential risk of aneuploid embryo and pre‐implantation genetic diagnosis or prenatal diagnosis.  相似文献   

2.
The aim of this work was to evaluate the use of air‐dried spermatozoa for in vitro production of equine embryos and verify if sperm extract activation and in vivo culture improve in vitro embryo production. Cooled spermatozoa (control) and air‐dried spermatozoa stored for 2, 14 or 28 days were used for ICSI sperm extract, or ionomycin was used for oocyte activation, and embryos were in vitro or in vivo (in mare′s oviduct) cultured for 7 days. With in vitro culture, cleavage rate was higher when activating with sperm extract (P < 0.05). No differences in embryo development were seen between the two activation treatments nor between storage periods (P > 0.05). Blastocysts were obtained with cooled spermatozoa, and morulae were achieved using in vivo culture with 28‐day storage spermatozoa and ionomycin‐activated oocytes. When in vivo culture was performed, sperm DNA fragmentation was assessed using the sperm chromatin dispersion test and did not show statistical correlation with cleavage nor embryo recovery rates. In conclusion, equine embryos can be produced using air‐dried spermatozoa stored for several weeks. Sperm extract activation increased cleavage rates but did not improve embryo development. In vivo culture allowed intrauterine stage embryos to be achieved.  相似文献   

3.
The method of choice for assisted reproductive technology treatment in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) is usually based on the evaluation of male infertility factors. Decisions for couples with moderate oligoasthenozoospermia (OA) are often empirical because uniform treatment criteria are lacking. This study aimed to evaluate the effect of patients with moderate OA treated with conventional IVF and ICSI. A total of 199 couples with moderate OA undergoing their first IVF/ICSI cycle were included in the study. The patients were divided into two groups according to the type of insemination: conventional IVF group (n = 97) and ICSI group (n = 102). All patients were randomised to be inseminated either by conventional IVF or ICSI. The fertilisation rate, embryo quality, implantation rate and clinical pregnancy rate were examined. No differences in the fertilisation, implantation and pregnancy rates were observed between conventional IVF and ICSI groups (> 0.05). However, the number of good‐quality embryos was significantly higher in the ICSI group than in the IVF group (P < 0.05). Couples with moderate OA did not influence on the overall clinical outcomes between IVF and ICSI treatments, and a negative influence by ICSI on blastocyst development was not confirmed.  相似文献   

4.
Excess of reactive oxygen species (ROS) on in vitro embryo production systems negatively affects the quality and developmental potential of embryos, as result of a decreased sperm quality and increased DNA fragmentation. This issue is of major importance in assisted fertilisation procedures such as intracytoplasmic sperm injection (ICSI), because this technique does not allow the natural selection of competent spermatozoa, and therefore, DNA‐damaged spermatozoa might be used to fertilise an egg. The aim of this study was to investigate a new strategy to prevent the potential deleterious effect of ROS on cryopreserved bovine spermatozoa. We evaluated the effect of a sperm pre‐treatment with different concentrations of N‐acetyl‐L‐cysteine (NAC) on ROS production, viability and DNA fragmentation and assessed the effect of this treatment on the in vitro developmental potential and quality of embryos generated by ICSI. The results show a strong scavenging effect of 1 and 10 mm NAC after exposure of spermatozoa to a ROS inducer, without compromising the viability and DNA integrity. Importantly, in vitro developmental potential and quality of embryos generated by ICSI with spermatozoa treated with NAC were not affected, confirming the feasibility of using this treatment before an ICSI cycle.  相似文献   

5.
Patients with severe spermatogenesis impairment can now successfully father a child thanks to the use of intracytoplasmic sperm injection (ICSI). In oligozoospermic patients, many studies have reported significantly higher sperm aneuploidy rates and therefore an increased risk of transmitting a chromosomal abnormality via the injection of abnormal spermatozoa. However, the frequency of aneuploidy is highly variable between patients. The aim of the present work was to identify clinical and biological factors, which, together with non-obstructive oligozoospermia, could be predictive of elevated sperm aneuploidies. The sperm aneuploidy rates for chromosomes X, Y, 13, 18 and 21 were assessed in 31 infertile men with well-characterized spermatogenesis impairment, and in a population of control men with proven fertility. The frequency of sperm aneuploidy was compared between several patient subgroups according to their clinical and biological factors. Nearly half of the oligozoospermic males (15/31) had a significantly increased disomy rate for at least one of the five chromosomes compared with that observed in the control population (mean disomy rates + 1.96 standard deviation). Factors significantly associated with higher numbers of aneuploid sperm were cigarette smoking, an elevated follicle-stimulating hormone level, a sperm concentration less than 1 m/mL, and a severe teratozoospermia. Hence, several factors predictive of an increased risk of sperm aneuploidy rates were identified in ICSI male candidates with a non-obstructive oligozoospermia.  相似文献   

6.
The purpose of this study was to compare the clinical outcome of testicular spermatozoon versus ejaculated spermatozoon in the treatment of infertile males with high sperm DNA damage, referred as sperm DNA fragmentation index (DFI), that attending intracytoplasmic sperm injection (ICSI) programme in terms of clinical pregnancy, births delivered as the primary and pregnancy loss and embryo fertilisation as the secondary outcome. A total of 102 males fulfilling the inclusion criteria were enrolled in the present study. Of the 102 males, 61 infertile males underwent testicular spermatozoon combined with ICSI while the remaining 41 males applied ejaculated spermatozoa in their first ICSI cycles, and the data of them were collected and analysed. In a 18‐month follow‐up, testicular spermatozoon achieved higher pregnancy rate and deliver rate than those in ejaculated sperm group (pregnancy rate, 36% vs. 14.6%, p = 0.017; deliver rate, 38.5% vs. 9.8%, p = 0.001). Nevertheless, there were no significant differences in the number of oocytes aspirated and number of embryos transferred between the two groups. Additionally, the fertilisation rate in the testicular sperm study cohort (70.4%) was also similar to that in the ejaculated sperm group (75.0%). Based on the current data, we conclude that testicular spermatozoon is the prior option in the treatment of infertile males with high sperm DFI in ICSI programme. More high‐quality studies with larger samples size are needed in the future due to the relative small size and the nonrandomized design of the present study.  相似文献   

7.
Various nuclear sperm alterations are reported in patients with syndromic teratozoospermia; however, this has not been clearly identified yet in total polymorphic teratozoospermia. The aim of this study was to analyse sperm aneuploidy, DNA integrity and chromatin packaging in 45 infertile patients with total polymorphic teratozoospermia, and to compare obtained results with those collected from 25 fertile men. For 14 patients, the impact of nuclear sperm abnormalities on intracytoplasmic sperm injection (ICSI) outcomes was analysed. Sperm chromatin condensation was evaluated using aniline blue staining, DNA fragmentation by TUNEL assay and chromosome abnormalities by FISH. The mean DNA fragmentation index was significantly higher in patients compared to controls, weakly and positively correlated to acrosome defects (r = 0.3; p = 0.04) and positively and moderately correlated to microcephalic heads (r = 0.5; p = 0.027). The aniline blue‐reacted spermatozoa rate was also high in comparison with controls, moderately and negatively correlated to progressive motility (r = ?0.6; p = 0.014). Total aneuploidy rate was considerably higher in our patients. A positive and moderate correlation was found between disomy Y rate and acrosome abnormalities (r = 0.5; p = 0.048). These patients had an impaired sperm nuclear quality, which will affect the results in ICSI. Therefore, analysis of sperm chromatin condensation, DNA integrity and aneuploidy in such cases is very useful before ART.  相似文献   

8.
In this study, our purpose was to investigate the possible effect of paternal obesity on intracytoplasmic sperm injection (ICSI) outcomes on the basis of clinical pregnancy outcome. Antropometric measurements of 155 couples, referred to our infertility clinic and who underwent an ICSI cycle, have been evaluated. The study sample were divided into three groups with respect to paternal body mass index (BMI), as normal weight (BMI: 20–24.9), overweight (BMI: 25–29.9) and obese (BMI ≥ 30). Results of conventional semen analysis were also analysed. Clinical pregnancy data, including fertilisation rate, implantation rate, clinical pregnancy rate and live birth rate, were evaluated. Paternal obesity was a significant negative factor for sperm concentration and sperm motility (P = 0.03 and P = 0.01 respectively). A significant decrease of clinical pregnancy rate and live birth rate was associated with increased paternal BMI (P = 0.04 and P = 0.03 respectively). We have not determined a significant difference among groups in terms of fertilisation rate and implantation rate. This study demonstrates that increasing paternal BMI has a negative influence on ICSI success, including clinical pregnancy rate and live birth rate. There is a need for further studies to point the importance of lifestyle changes in order to overcome the negative influence of paternal obesity on couple's fertility.  相似文献   

9.
Sperm aneuploidy is a result of mis-segregation during meiosis and correlates with male infertility. Among the types of aneuploidy, nullisomy has been reported to be more prevalent in human spermatozoa than disomy; however, nullisomy is not always assessed by FISH, and its relation with basic semen parameters is almost unknown. To establish an association between nullisomy and semen parameters and pathologies, we evaluated the potential clinical value of semen analysis and assessed the diagnosis of sperm nullisomy. A prospective study including a total of 130 patients and 25 donors aged 30–50 years with a normal karyotype was carried out. Sperm FISH analyses were performed, and basic semen parameters and ART outcome data were collected. There were no associations between sperm nullisomy of chromosomes 13, 15, 18, 21, 22, X and Y and basic semen parameters. The odds of nullisomy of chromosomes 13, 15, 16, 17, 18, 21, 22, X and Y were not related to semen pathologies. However, sperm nullisomy had a negative impact on ART outcomes, with significant decreases in fertilisation, blastocyst, pregnancy and implantation rates after ICSI. Sperm nullisomy diagnoses are not detected in semen analyses and are a possible cause of idiopathic male infertility and unexplained recurrent pregnancy loss.  相似文献   

10.
Basic semen analysis is insufficient for determining the fertility potential. The aim of this study was to determine if hyperactivated motility (HAM) and acrosome reaction (AR) can be useful tests for evaluating semen quality during male infertility evaluations and to help the clinician decide whether regular insemination or intracytoplasmic sperm injection (ICSI) is preferable during in vitro fertilisation. A prospective study was conducted. Patients with normal sperm according to World Health Organization guidelines who underwent IVF treatment and planned regular insemination were asked to participate. A portion of sperm sample was evaluated for HAM and AR on day of ovum pick up. In HAM assessment, 93.3% of patients with increased HAM had a high fertilisation rate compared with 64% in the group without increased HAM (P = 0.059). For the AR evaluation, 91.7% of samples with a low rate of spontaneous AR had a high fertilisation rate compared with 39.3% in the group with a high rate of spontaneous AR (= 0.004).  相似文献   

11.
The objective of this study was to determine the aneuploidy rate in spermatozoa from a globozoospermic patient who underwent three unsuccessful intracytoplasmic sperm injection (ICSI) treatment cycles together with his wife. The aneuploidy rates of chromosomes 13, 18, 21, X and Y as well as the diploidy rate in the spermatozoa of this man were evaluated by fluorescence in situ hybridisation to clarify if chromosomal aneuploidy could be a cause for the low fertilisation rate observed in this case. In the spermatozoa of our patient no increase in aneuploidy rates was found for the tested chromosomes (0.0% disomies of chromosomes 13, 18 and 21 with a diploidy rate of 0.6%; X/Y missegregation: 0.6%) indicating that the ICSI outcome of the couple was not impaired by sperm aneuploidy of the globozoospermic man.  相似文献   

12.
Low sperm quality has negative effects on fertilisation and embryo development. The males with azoospermia apply for testicular sperm extraction (TESE) or microsurgical epididymal sperm aspiration (MESA) in order to retrieve sperm. To date, there have not been any reports investigating morphokinetic parameters of pre‐implantation embryos using testicular and epididymal spermatozoa. Therefore, we aimed to correlate embryo development and assess morphogenetic parameters in embryos obtained by TESE and MESA using time‐lapse imaging. A total of 60 patients undergoing IVF treatments were included in this study. Twenty men with normal semen parameters were selected as control group. Twenty men undergoing TESE and 20 men undergoing MESA were also included in this study. The morphokinetic parameters of time intervals between the second polar body (PB2) extrusion, pronuclei formation and disappearance and cleavage divisions showed significant variations in TESE, MESA and control groups. Furthermore, the pregnancy rates (positive beta‐hCG) were shown to be similar in both TESE and the control group (55% in each group), whereas for the MESA group, this rate was significantly lower (39%, p = 0.049). Further extrapolation of these results may implicate that the obstructive azoospermia patients should undergo TESE instead of MESA for better blastocyst development and higher pregnancy rates.  相似文献   

13.
Ubiquitin is an 8.5‐kDa protein that tags outlived proteins for degradation by the proteasome. It also marks defective spermatozoa during epididymal passage and has been proposed as a biomarker of sperm quality. This study evaluates the relationship between sperm ubiquitination, protamine deficiency, semen parameters and fertilisation rate in infertile individuals undergoing the intracytoplasmic sperm insemination (ICSI) procedure. Semen samples from 73 ICSI candidates were collected and analysed according to World Health Organization criteria. A portion of each sample was evaluated for sperm ubiquitination using the sperm ubiquitin tag immunoassay (SUTI) with flow cytometry, and protamine deficiency by chromomycin A3 (CMA3) staining. In addition, the relationship between the fertilisation rate and sperm ubiquitination was calculated in ICSI candidates. The intensity of ubiquitination showed a significant negative correlation with sperm concentration (r = ?0.255, P = 0.032) and a positive correlation with fertilisation rate (r = 0.384, P = 0.013) post‐ICSI. No correlation was observed between protamine deficiency and the percentage of ubiquitination or ubiquitination intensity. The results of this study suggest that sperm ubiquitination prior to capacitation may be considered as a marker of defective spermatozoon. Spermatozoa that undergo proper ubiquitination may have a higher chance for fertilisation, because they are made redundant by the ubiquitin–proteasome pathway in the epididymis compared to hypo‐ubiquitinated spermatozoa.  相似文献   

14.
Zona pellucida (ZP)-bound spermatozoa have normal morphology and motility and can enhance the ICSI outcomes. Selection of zona pellucida-bound spermatozoa is recently considered to find functional spermatozoa for ICSI. This study reviewed the efficacy of ZP-bound sperm selection on the ICSI outcomes includes fertilisation rate, embryo quality, embryo transfer rate and clinical pregnancy rate. The databases searched include PubMed, Scopus and Cochrane databases up to January 2019. All research reports with full text and in English language that addressing the relation between ZP-sperm selection and ICSI outcomes were included. Fifty studies were suitable after screening of the 845 identified articles. After exclusions, five of these studies were included. Meta-analytic pooling of data indicated no association between the ICSI outcomes and ZP-bound sperm selection except a marginal effect on implantation rate. Eliminating one study indicated that ZP-bound sperm selection technique improves embryo quality, implantation rate and clinical pregnancy rate. This study revealed that ZP-bound sperm selection produces only a slight improvement in implantation rate. However, further studies with a large number of couples must be done to clarify the potential beneficial effect of ZP-bound spermatozoa on ICSI outcomes.  相似文献   

15.
The aim of this study was to compare the sperm morphology and nuclear sperm quality (sperm aneuploidy and DNA fragmentation) in two groups of globozoospermic patients: DPY19L2‐mutated patients (n = 6) and SPATA16‐mutated patients (n = 2). Results for these two groups were also compared to a group of fertile men (n = 25). Fluorescence in situ hybridisation was performed for chromosomes X, Y and 18. Sperm DNA fragmentation was evaluated by TUNEL assay. Sanger sequencing was performed for mutations screening of DPY19L2 and SPATA16 genes. Sperm analysis revealed a classic phenotype of total globozoospermia in DPY19L2‐mutated group and a particular phenotype characterised by a predominance of double/multiple round‐headed (39.00 ± 4.2%) and multi‐tailed spermatozoa (26.00 ± 16.97%) in SPATA16‐mutated group. FISH analysis showed a significantly higher aneuploidy rate in globozoospermic patients compared to controls (p < 0.05), and a higher rate was observed in SPATA16‐mutated group compared to DPY19L2‐mutated group (p < 0.05). DNA fragmentation index was significantly higher in globozoospermic men compared to controls (p < 0.001), and there is no statistically significant difference between the two globozoospermic groups. We showed that SPATA16 defects could be associated with an abnormal meiosis leading to a particular morphological sperm defect of double/multiple round‐headed and multi‐flagella and a higher sperm aneuploidy rate than in case of DPY19L2‐defects in classic globozoospermia.  相似文献   

16.
C. Zhao  L. Zhou  J. Zhang  X. Ling 《Andrologia》2016,48(10):1138-1144
To explore the need for rescue intracytoplasmic sperm injection (ICSI) in cases of partial fertilisation failure during a conventional in vitro fertilisation cycle, rescue ICSI was performed for cycles with a fertilisation rate of <50%. The data were divided into three groups based on the fertilisation rate: group 1 (0%), group 2 (<25%) and group 3 (>25%). The impact of rescue ICSI on each group was then analysed in terms of ovum fertilisation, embryo development, embryo utilisation and selection of embryos for transfer. Rescue ICSI was performed on 1831 unfertilised oocytes from 313 cycles. The fertilisation rates for group 1, group 2 and group 3 were 74.66, 68.35 and 65.46%, and the rate of polyploidy in the three groups was 8.55, 11.33, and 14.47%. The percentage of embryos that can be transferred from rescue ICSI for group 2 was 38.33%, and this value was higher than those of the other two groups. It is concluded that rescue ICSI is not recommended for patients with an IVF rate of >25% as the procedure is associated with a greater risk and low returns. However, it is feasible to perform rescue ICSI for patients with IVF rates of <25%.  相似文献   

17.
Oxidative stress has negative impacts on the clinical outcomes of assisted reproduction techniques. The brain‐derived neurotrophic factor (BDNF) promotes the viability of nerve cells and is known to decrease oxidative stress and apoptosis in different cells. The aim of this study was to evaluate the effect of BDNF treatment on human sperm functions that are known to be essential for fertilisation. Our findings showed that treatment of human spermatozoa with 0.133 nM BDNF significantly increased the percentages of both total (= 0.001) and progressive (P < 0.01) motile sperm cells compared to those observed in the nontreated (control) group. We also showed that the mean fluorescence intensity of DCFH‐DA, as an indicator of intracellular reactive oxygen species, was significantly lower (P < 0.05) in spermatozoa treated with BDNF compared to the control group. Treatment of spermatozoa with BDNF significantly decreased the percentages of both dead (= 0.001) and apoptotic‐like sperm cells (P < 0.05) compared to the control group. On the other hand, BDNF treatment significantly increased the percentage of viable sperm cells compared to the control (= 0.001). In conclusion, BDNF has protective effects against oxidative stress in spermatozoa and could improve sperm functions that are essential for sperm–egg fusion and subsequent fertilisation.  相似文献   

18.
Multiple morphological abnormalities of the sperm flagella (MMAF) are a rare type of male infertility. Mutations in DNAH1, CFAP43 and CFAP44 are the main aetiology of the disorder. Previously, good intracytoplasmic sperm injection (ICSI) outcomes were reported for MMAF patients with DNAH1 mutations. However, the ICSI prognosis for MMAF patients with CFAP43 or CFAP44 mutations was not known. We designed a retrospective cohort study. Molecular genetic testing identified six MMAF patients with biallelic CFAP44 (CFAP44+ group) or CFAP43 mutations and 12 patients with homozygous or compound heterozygous DNAH1 mutations (DNAH1+ group). A control group consisted of age‐matched, non‐MMAF men. For MMAF patients carrying CFAP44 mutations, the recorded rates of fertilisation, transferable embryos, pregnancy and delivery after ICSI were 76.47%, 88.46%, 50.0% and 50.0% respectively. The fertilisation rate was significantly higher in the CFAP44+ group than in the DNAH1+ group (76.47% vs. 54.5%, p = 0.0196). There were no statistically significant differences in the rates of transferable embryos, implantation, clinical pregnancy and miscarriage between the CFAP44+ group and either the DNAH1+ group or the age‐matched control group. Our results support a good ICSI prognosis for MMAF patients carrying CFAP44 or CFAP43 mutations.  相似文献   

19.
The main purpose of this prospective study is to examine possible influences of abnormalities of sperm nuclear condensation and chromatin decondensation with sodium dodecyl sulphate (SDS)‐EDTA on outcomes of intrauterine insemination (IUI) or intracytoplasmic sperm injection (ICSI) cycles. Semen samples from 122 IUI and 236 ICSI cycles were evaluated. Before semen preparation for IUI or ICSI, basic semen analysis was performed and a small portion from each sample was spared for fixation. The condensation of sperm nuclear chromatin was evaluated with acidic aniline blue, followed by sperm chromatin decondensation by SDS‐EDTA and evaluation under light microscope. Ongoing pregnancy rate was 24% and 26.2% in the IUI and ICSI groups respectively. The chromatin condensation rate was significantly higher in the ongoing pregnancy‐positive group compared to the negative group, both in IUI (P = 0.042) and ICSI groups (P = 0.027), and it was positively correlated with ongoing pregnancy rate in both IUI and ICSI groups (P = 0.015, r = 0.214 and P = 0.014, r = 0.312 respectively). Chromatin decondensation rates were not significantly different in neither of the groups. These results indicate that IUI and ICSI outcome is influenced by the rate of spermatozoa with abnormal chromatin condensation. Sperm chromatin condensation with aniline blue is useful for selecting assisted reproduction techniques (ART) patients.  相似文献   

20.
We investigated whether the presence of Y chromosome azoospermia factor (AZF) microdeletions impacts upon the outcomes of intracytoplasmic sperm injection (ICSI) using fresh ejaculated spermatozoa. Sixteen oligozoospermia patients with Y chromosome AZFb or AZFc microdeletions and undergoing ICSI cycles between March 2013 and November 2014 were studied. Twenty‐six infertile men with normal Y chromosomes and also undergoing IVF/ICSI in the same time period were used as controls. A retrospective case–control study approach was used. Among the 16 cases, 12 (75%, 12/16) had deletions of AZFc markers (sY152, sY254 and sY255), one (6.25%, 1/16) had a deletion of sY152, and two (12.5%, 2/16) had deletions of sY152, sY254, sY255 and sY157. AZFb microdeletions were found in one patient (6.25%, 1/16). There were no significant differences between groups for cleaved embryo rate, high‐grade embryo rate, blastocyst formation rate, embryo implantation rate, clinical pregnancy rate and delivery rate. The clinical outcomes of ICSI for oligozoospermic patients with Y chromosome AZF microdeletion are comparable to those of infertile patients with normal Y chromosomes. Our findings indicate that ICSI should be offered to patients with an AZFc deletion and that oligozoospermia patients with AZFb microdeletions are likely to father children.  相似文献   

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