scholarly journals A Simple, Centrifugation-Free, Sperm-Sorting Device Eliminates the Risks of Centrifugation in the Swim-Up Method While Maintaining Functional Competence and DNA Integrity of Selected Spermatozoa

2020 ◽  
Vol 28 (1) ◽  
pp. 134-143
Author(s):  
Huidrom Yaiphaba Meitei ◽  
Shubhashree Uppangala ◽  
Krishna Sharan ◽  
Srinidhi Gururajarao Chandraguthi ◽  
Arunkumar Radhakrishnan ◽  
...  
Keyword(s):  
Acrosome Reaction ◽  
Semen Analysis ◽  
Dna Lesions ◽  
Dna Integrity ◽  
Sperm Selection ◽  
Mitochondrial Integrity ◽  
Sperm Dna ◽  
Radiation Induced ◽  
Normal Head ◽  
Intra Uterine Insemination

AbstractThis pilot study was conducted to explore the benefits of using a centrifugation-free device based on the migration–sedimentation (MS) technique over centrifugation-based techniques in selecting competent spermatozoa, as compared with using split human semen samples. Ejaculates from 35 men undergoing semen analysis were split into four parts where one part was retained as the neat (NE) and the other three parts were subjected to sperm selection by using migration–sedimentation (MS), density gradient (DG) separation, and swim-up (SU) techniques. Sperm functional characteristics along with mitochondrial integrity, tyrosine phosphorylation, acrosome reaction, and ultrastructure were measured. The ability of selection techniques in reducing spontaneous and radiation-induced sperm DNA lesions was assessed by the TUNEL assay. In results, MS-selected spermatozoa had higher viability (P < 0.001), longevity in terms of total motility at the end of 6 and 18 h post-extraction (P < 0.001), and mitochondrial integrity (P < 0.001) compared with those selected by DG. Furthermore, spontaneous DNA lesions were significantly reduced in MS and SU fractions compared with NE (P < 0.001). Similarly, radiation-induced sperm DNA lesions were significantly lower in MS and SU fractions (P < 0.001) compared with DG. Ultrastructural analysis using scanning electron microscopy suggested a moderate, non-significant increase in the number of spermatozoa with normal head and mid-piece in MS fraction compared with other methods. In conclusion, the MS-based device offers a centrifugation-free, efficient, and reliable sperm selection method, making it suitable for partially equipped intra-uterine insemination (IUI) laboratories or office IUI programmes. Further research should focus on the safety and clinical usefulness of the device in assisted conception programmes in general and IUI in specific.

3 SELECTION OF UNFRAGMENTED-DNA SPERMATOZOA FROM HEAT STRESSED MICE BY FEMALE UTERINE TRACT AND ZONA PELUCIDA BINDING

2009 ◽  
Vol 21 (1) ◽  
pp. 102
Author(s):  
J. D. Hourcade ◽  
M. Perez-Crespo ◽  
B. Pintado ◽  
A. Gutiérrez-Adán
Keyword(s):  
Heat Treatment ◽  
Dna Damage ◽  
Sperm Motility ◽  
Tail Length ◽  
Dna Integrity ◽  
Sperm Selection ◽  
Epididymal Sperm ◽  
Cleavage Rate ◽  
Sperm Dna ◽  
Selection Of

Physiological bases of the sperm selection processes within the female reproductive tract before they meet and fertilize the oocyte are unknown. The aim of this work was to determine if one of the keys of spermatozoa selection could be DNA integrity. It has been reported that sperm DNA damage does not impair in vitro fertilization (IVF). However, it has been suggested that the zona pelucida (ZP) is able to select spermatozoa with unfragmented DNA (Liu and Baker 2007 Hum. Reprod. 22, 1597–1602). In this work, DNA damage of spermatozoa was artificially induced by scrotal heat treatment (HT) (42°C, 30 min). Twenty-one days after the HT, spermatozoa were recovered from the epididymis caudae of CD1 mice and from the uterine horns near the cervix (Uc), from the uterine horns near the oviducts (Uo), and from the oviducts (Ov) of CD1 females 1–2 h after mating with HT and control males. In each region we determined numbers of spermatozoa, individual motility and sperm DNA integrity by COMET assay (% DNA in tail, tail length, and COMET moment was calculated). Also, females naturally mated either with HT or control males were killed at Day 14 of pregnancy, and number of foetuses and resorptions was recorded. Additionally, IVF was performed with epididymal sperm from HT or control males, Two hours after IVF attached and un-attached spermatozoa to the ZP were recovered and samples were evaluated for sperm motility (CASA), sperm zona-binding, and sperm DNA fragmentation (COMET). Also cleavage rate of fertilized oocytes with sperm from HT or control males was analyzed. One-way ANOVA was used to compare the results form each group. Epididymal sperm count (12*106 and 4.4*106 for control and HT respectively), sperm motility (75 and 21% respectively) and testis weight (133.90 and 68.76 mg, respectively) were significantly reduced after heat treatment (P < 0.001). For the heat treatment, COMET values decreased significantly during the transit from Uc to Uo and from Uo to Ov (Tail DNA: 25.7, 23.5, and 14.4% respectively, P < 0.01; Tail length: 38.4, 29.4, and 11.2 pixels, P < 0.001; COMET Moment: 12.5, 8.5, and 2 respectively, P < 0.001). Heat treatment reduced numbers of foetuses (7 ± 0.5 v. 5 ± 0.49, control and HT group, respectively), but number of resorptions was not altered. Spermatozoa bound per ZP in IVF experiments (55 ± 7 and 13 ± 6, control and HT, respectively) and cleavage rate (61 ± 1 v. 15 ± 6, control and HT, respectively) were significantly reduced in the HT group. Two hours after IVF, spermatozoa attached to the ZP in HT group showed a significant decrease in COMET parameters as in tail length (59.46 ± 2.895 v. 34.66 ± 3.531), and in tail moment compared with unattached spermatozoa. Our results indicate that DNA integrity sperm selection mechanisms are present in both the female tract and the ZP. We suggest that genital tract and sperm-ZP binding process plays an important role in selection of sperm with normal chromatin DNA.

Should sperm DNA integrity be studied in the first semen analysis? A comparison between etiological factors

2013 ◽  
Vol 100 (3) ◽  
pp. S437-S438
Author(s):  
A. Mata ◽  
M.B. Falco ◽  
G. Gustavo ◽  
A.M. Juárez Villanueva ◽  
C.A. Sánchez Sarmiento ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Dna Integrity ◽  
Etiological Factors ◽  
Sperm Dna Integrity ◽  
Sperm Dna

Sperm DNA fragmentation: causes and identification

Zygote ◽  
2019 ◽  
Vol 28 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Thais Rose dos Santos Hamilton ◽  
Mayra Elena Ortiz D’Ávila Assumpção
Keyword(s):  
Dna Fragmentation ◽  
Semen Analysis ◽  
Sperm Chromatin ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Functional Capability ◽  
Sperm Dna Integrity ◽  
Sperm Dna ◽  
Fertility Disorders ◽  
Chromatin Integrity

SummarySperm DNA fragmentation is referred to as one of the main causes of male infertility. Failures in the protamination process, apoptosis and action of reactive oxygen species (ROS) are considered the most important causes of DNA fragmentation. Action of ROS or changes in sperm protamination would increase the susceptibility of sperm DNA to fragmentation. Routine semen analysis is unable to estimate sperm chromatin damage. Sperm DNA integrity influences sperm functional capability, therefore tests that measure sperm DNA fragmentation are important to assess fertility disorders. Actually, there is a considerable number of methods for assessing sperm DNA fragmentation and chromatin integrity, sperm chromatin stability assay (SCSA modified), sperm chromatin dispersion (SCD), comet assay, transferase dUTP nick end labelling (TUNEL); and protamine evaluation in sperm chromatin assay, such as toluidine blue, CMA3, protamine expression and evaluation of cysteine radicals. This review aims to describe the main causes of sperm DNA fragmentation and the tests commonly used to evaluate sperm DNA fragmentation.

Acrosome reaction after ionophore challenge: relationship to sperm DNA integrity

2004 ◽  
Vol 1271 ◽  
pp. 197-199 ◽  
Author(s):  
S. Moskovtsev ◽  
J. Willis ◽  
A. Azad ◽  
B. Mullen
Keyword(s):  
Acrosome Reaction ◽  
Dna Integrity ◽  
Sperm Dna Integrity ◽  
Sperm Dna

scholarly journals Relationships between sperm DNA integrity and bulk semen parameters in Bulgarian patients with varicocele

2019 ◽  
Vol 91 (2) ◽  
Author(s):  
Viktor Alargkof ◽  
Larissa Kersten ◽  
Romil Stanislavov ◽  
Zdravko Kamenov ◽  
Panagiotis Nikolinakos
Keyword(s):  
Dna Fragmentation ◽  
Semen Analysis ◽  
Human Reproduction ◽  
Sperm Parameters ◽  
Semen Parameters ◽  
Control Group ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Abnormal Sperm ◽  
Sperm Dna

Objective: This exploratory retrospective study aimed to compare the level of Sperm DNA Fragmentation (SDF) and investigate its association with bulk semen parameters, for the first time in Bulgarian patients with varicocele, using a distinct methodology. Material and methods: Standard semen analysis was performed according to the 2010 criteria of the European Society of Human Reproduction and Embryology - Nordic Association for Andrology (ESHRE-NAFA-2010) and DNA fragmentation was assessed using the Halosperm® kit. The total sample included 28 males: the control group consisted of men with normal genital examination and unknown fertility (n = 10), group one consisted of men with varicocele, normozoospermia and DNA fragmentation > 15% (n = 9) and group two consisted of men with varicocele, abnormal sperm parameters and DNA fragmentation > 15% (n = 9). Results: DNA fragmentation was found to be higher in patients with abnormal sperm parameters (43.78 ± 30.78) compared to the normozoospermic group (21.22 ± 3.93) (p = 0.008). In normozoospermic patients, no statistically significant correlations were observed between SDF and bulk semen parameters. In patients with abnormal sperm parameters, DNA fragmentation exhibited significant very strong negative association with motility (a+b), vitality and typical morphology (p < 0.001). Conclusions: DNA integrity assays could be used for a better evaluation and management of male infertility, particularly in normozoospermic varicocele patients.

scholarly journals Sperm DNA Integrity Test and Assisted Reproductive Technology (Art) Outcome

2016 ◽  
Vol 9 (1) ◽  
pp. 21-29 ◽  
Author(s):  
Violeta S. Rilcheva ◽  
Nina P. Ayvazova ◽  
Lyubomira O. Ilieva ◽  
Svetlana P. Ivanova ◽  
Emiliana I. Konova
Keyword(s):  
Pregnancy Rate ◽  
Assisted Reproductive Technology ◽  
Dna Fragmentation ◽  
Intrauterine Insemination ◽  
Semen Analysis ◽  
Reproductive Technology ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Integrity Test ◽  
Sperm Dna

SummaryThe objective of the study was to investigate the influence of sperm DNA fragmentation index (DFI) by DNA integrity test on pregnancy outcome and pregnancy loss after assisted reproductive technology (ART) procedure: autologous intracytoplasmic sperm injection (ICSI), donation eggs ICSI, and intrauterine insemination (IUI). We investigated men from 531 couples undergoing autologous ICSI procedure (n=416), from couples undergoing donation eggs procedure (n=39) and IUI (n=76). We performed the following interventions: semen analysis, DNA integrity test, embryo scoring by Gardner and Schoolcraft grading system (1999). The study showed no statistically significant differences between the group regarding pregnancy rate (χ2=0.55; p>0.05; OR=1.25, 95% Cl 1.23-1.46; p>0.05). However, with increased levels of DFI, the number of pregnancy losses became higher (including biochemical pregnancies and spontaneous abortions) at OR=5.65 (95% Cl 4.32-7.11; p=0.05). We examined the percentage of grade I blastocysts (by Gardner and Schoolcraft, 1999) before donation eggs embryo transfer and found a statistically significant correlation with both the DFI (χ2=7.80; p<0.05) and sperm morphology (χ2=6.14; p<0.05). Analysis of the relationship between DFI and IUI output (clinical pregnancy, miscarriage) revealed significant correlations in both directions: between DFI and pregnancy rate after IUI (χ2=6.29; p<0.05) and between the DFI and pregnancy development after IUI (χ2=6.87; p<0.05). The three group categories (autologous, heterologous ICSI procedures and IUI) studied showed that sperm samples with DFI>27% were associated with increased riskofearlypregnancyloss. Men with infertility should undergo DNA fragmentation assay in addition to the standard semen analysis. When DFI exceeds 27%, ICSI should be a method of choice, even in cases where the conventional parameters of semen analysis tests are normal.

Clinical Comparison of Traditional Semen Analysis Parameters and Sperm DNA Integrity Assays.

2008 ◽  
Vol 78 (Suppl_1) ◽  
pp. 226-226
Author(s):  
Dennis Marchesi ◽  
Hannah Biederman ◽  
Y. E. Hung ◽  
J. Tai ◽  
A. Hershlag ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Dna Integrity ◽  
Sperm Dna Integrity ◽  
Clinical Comparison ◽  
Sperm Dna

scholarly journals Normal seminal plasma could preserve human spermatozoa against cryopreservation damages in Oligozoospermic patients

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Fatemeh Eini ◽  
Maryam Azizi kutenaei ◽  
Maryam Hosseinzadeh Shirzeyli ◽  
Zeinolabedin Sharifian Dastjerdi ◽  
Mahmoud Omidi ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Semen Analysis ◽  
Human Sperm ◽  
Human Spermatozoa ◽  
Dna Integrity ◽  
Fresh Sample ◽  
Sperm Characteristics ◽  
Sperm Dna ◽  
Plasma Effects ◽  
And Function

Abstract Background Cryopreservation of human spermatozoa has been identified as an efficient procedure to preserve fertility in men before any cancer therapy or surgical infertility treatment. Despite the benefits of the procedure, the deleterious effects of cryopreservation have been proven on sperm structure and function. This study aimed to evaluate seminal plasma effects on human sperm characteristics after cryopreservation, and compare the addition of normozoospermic and oligozoospermic seminal plasma in the prepared oligozoospermic samples. Semen samples were collected from fifty-five oligozoospermic men and the twenty fertile individuals who referred to the infertility center. At first, a semen analysis was carried out on each neat ejaculate, and then some were cryopreserved. The remainder of the semen was divided into two, one for seminal plasma removal and the other for sperm preparation. Then, the prepared spermatozoa were cryopreserved in three groups: one with, and another without the addition of oligozoospermic seminal plasma, and still another with the addition of normal seminal plasma. After thawing, sperm DNA integrity, viability, motility, and morphology were determined. Results The percentages of all parameters were significantly lower after cryopreservation in all groups compared to the fresh sample. However, this reduction was lower in the oligozoospermic samples cryopreserved with normal seminal plasma. Conclusion The results indicated that seminal plasma in oligozoospermic patients could not support sperm against cryo-injuries, an indication likely due to insufficient antioxidants and other protective components in oligozoospermic patients. However, normal seminal plasma could slightly preserve sperm characteristics after cryopreservation in oligozoospermic patients.

scholarly journals Sperm morphology and DNA fragmentation after zona pellucida selection

2021 ◽  
Author(s):  
Rumiana Ganeva ◽  
Dimitar Parvanov ◽  
Denitsa Velikova ◽  
Magdalena Vasileva ◽  
Kristina Nikolova ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Zona Pellucida ◽  
Processing Technique ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Sperm Selection ◽  
Normal Morphology ◽  
Sperm Dna ◽  
Morphological Defects ◽  
The Mean

Sperm DNA fragmentation (SDF) and sperm morphological defects can negatively affect ART outcomes. Consequently, there is a need for additional semen processing technique that accounts for sperm DNA status and morphology prior to ICSI. The object was to evaluate the efficacy of an additional zona pellucida adhesion based sperm selection for obtaining sperm populations with high percentage of normal morphology and DNA integrity as compared to native semen and routine swim-up preparation. Semen samples from 78 normozoospermic men were subjected to swim up and placed in petri dishes coated with 48 acid-solubilised zonae pellucidae. Sperm DNA fragmentation and morphology were assessed in the native semen, the swim-up samples and the zona-adhered spermatozoa from each patient. The mean sperm DNA fragmentation of the zona-selected spermatozoa (3.5±0.7%) were significantly lower than the swim-up samples (15.3±5.2%) (p<0.001) and native semen (24.9±7.1%) (p<0.001). All of the samples had lower levels of DNA damage after additional selection by zona pellucida adhesion. Significantly higher percentage of sperm with normal morphology was observed after zona-adhesion selection (11.4±3.9%) when compared to the swim-up samples (8.9±4.3%) (p<0.001) or the native semen (5.3±3.2%) (p<0.001). In 94% of the samples, the percentage of spermatozoa with normal morphology increased after the additional zona selection. The present study demonstrates that sperm selection by additional zona-adhesion technique yields a significantly higher percentage of spermatozoa with normal morphology as well as a significantly decreased level of DNA fragmentation when compared to the native semen and the swim-up-only prepared samples.

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