P–049 The presence of double heads is associated with increased frequency of the other sperm abnormalities

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Handzhiyska ◽  
D Parvanov ◽  
R Ganeva ◽  
D Aleksandrova ◽  
E Tascudi ◽  
...  
Keyword(s):  
Normal Forms ◽  
Negative Impact ◽  
The Other ◽  
Frequency Of Occurrence ◽  
Vitro Fertilization ◽  
Multiple Defects ◽  
Sperm Abnormalities ◽  
Registration Number ◽  
Morphological Defects

Abstract Study question Is there an association between the presence of spermatozoa with double heads and the other sperm abnormalities in human semen? Summary answer Patients with double-headed spermatozoa had a significantly increased percentage of morphological abnormalities (head, midpiece and tail defects). What is known already The morphological evaluation of spermatozoa has a prognostic value for successful IVF procedure. It has been proven that certain morphological defects have a negative impact on fertilization, embryo quality, and pregnancy outcome in in-vitro fertilization cycles. Sperm abnormalities, such as double head, double tail and thin midpiece are rarely observed. However, their effect on the other sperm defects has not been well studied yet. The aim of this study was to examine the effect of the presence of double-headed spermatozoa on the frequency of occurrence of the other sperm defects. Study design, size, duration This retrospective study includes 2140 men aged between 18 and 73 years, with a mean of 36 years. It was conducted at Nadezhda Women’s Health Hospital, Bulgaria between October 2015 and August 2020. A comparative analysis was performed between semen samples with and without double-headed spermatozoa and the other sperm abnormalities, as well as the percentage of morphologically normal forms. Participants/materials, setting, methods Morphological analysis was performed according to the Kruger’s strict criteria. Totally 23 types of abnormalities were determined: head defects (small, large, amorphous, elongated, round, pear-shaped, double, acephalic, detached head, small and large acrosomal areas and spermatozoa without acrosome), midpiece defects (thick, bent, asymmetric, thin midpiece and cytoplasmic droplets), tail defects (stumped, coiled and double tail), acrosomal vacuoles, nuclear vacuoles and multiple defects. Statistics: Mann-Whitney U-test and T-test; P ≤ 0.05. Main results and the role of chance Presence of double-headed spermatozoa was observed in 12.62% (270/2140) of the studied samples. In these patients the frequency of occurrence of double-headed spermatozoa ranged between 1% and 29% with a mean of 0.41%±1.71%. Men with double-headed spermatozoa had significantly higher percentage of spermatozoa with small heads (24.51%±22.65%, P = 0.04), round heads (11.69%±10.13%, P < 0.01), nuclear vacuoles (10.64%±5.25%, P < 0.01), sperm without acrosome (9.76%±8.61%, P = 0.05), asymmetric midpiece (4.73%±3.96%, P < 0.05), bent midpiece (8.9%±7.22%, P < 0.01), thin midpiece (2.13%±4.44%, P < 0.01), double tail (1.78%±0.8%, P < 0.01), detached head (1.98%±1.42%, P < 0.01), stumped tail (6.03%±5.19%, P = 0.02), and cytoplasmic droplets (8.86%±5.02%, P < 0.01) compared to the patients without double-headed spermatozoa. Moreover, the percentage of sperm with multiple defects in the double-headed group was significantly higher (35.53%±29.91%, P < 0.01), while the percentage of normal forms was significantly lower (2.93%±3.64%, P < 0.01) compared to the patients without double heads. Limitations, reasons for caution In this study unequal sample sized groups were compared. We also need to investigate whether the obtained results will be confirmed in patients with certain pathological states, such as oligozoospermia, teratozoospermia, and asthenozoospermia. Wider implications of the findings: The present study revealed that the presence of double-headed spermatozoa in the ejaculate is related to an increased frequency of the other semen abnormalities. The double-headed spermatozoa could be used as an indicator for the total morphological quality of human spermatozoa Trial registration number Not applicable

scholarly journals Success rate of intrauterine insemination in patients with unknown infertility

2012 ◽  
Vol 69 (4) ◽  
pp. 301-307 ◽  
Author(s):  
Vladimir Jasovic ◽  
Emilija Jasovic-Siveska
Keyword(s):  
Success Rate ◽  
Negative Impact ◽  
Intrauterine Insemination ◽  
Unexplained Infertility ◽  
Mild Form ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B ◽  
Twin Pregnancies

Background/Aim. Unknown cause of infertility exists in 10%-26% of couples with infertility problems. Treatment of these couples depends on the possibility of correcting the unidentified defect over time. Intrauterine insemination (IUI) and ovaluation stimulation are methods of choice in treatment of unexplained fertility, but if a woman is older than 37 years, in vitro fertilization (IVF) could be directly recommended. The aim of this research was to compare the success rate of pregnancies with IUI between the patients with unexplained infertility and the patients with mild form endometriosis. Methods. The study included on 50 patients diagnosed with mild form endometriosis (group A) and 50 patients with unknown cause infertility (group B). Using the same therapeutical protocol, human menopausal gonadothropin (hMG) stimulation and horionic gonadropin (hCG) induction were applied, as well as IUI. Results. The percentage of achieved ovulation was higher in the group B (p < 0.05). During the 3 simulated sequential periods 102 IUI were performed in the group A and 97 IUI in the group B. In the group A there were 6 single and 1 twin pregnancies sucesfully conceived (14%), while in group B there were 9 (18%) single pregnancies. Conclusion. The use of a combination of controled ovarian hyperstimulation and IUI is an effective, cheap and safe method for treating infertility couples, especially couples with unknown cause infertility. Mild form endometriosis, as etiological infertility factor, has a negative impact on IUI success rate.

scholarly journals Using Time Lapse Monitoring for Determination of Morphological Defect Frequency in Feline Embryos after in Vitro Fertilization (IVF)

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 3 ◽  
Author(s):  
Barbara Kij ◽  
Joanna Kochan ◽  
Agnieszka Nowak ◽  
Wojciech Niżański ◽  
Sylwia Prochowska ◽  
...  
Keyword(s):  
Time Lapse ◽  
Blastocyst Stage ◽  
Hatching Rate ◽  
Vitro Fertilization ◽  
First Results ◽  
Morphological Defects ◽  
Defect Frequency ◽  
Selection Of

Some human, bovine, and mouse in vitro fertilized (IVF) embryos with morphokinetic abnormalities such as fragmentation, direct cleavage, and cytoplasmic vacuoles have the potential to reach the blastocyst stage, which is related to a high potential for implantation. The latest techniques of embryo development observation to enable the evaluation and selection of embryos are based on time lapse monitoring (TLM). The aim of this study was to determine the frequency of morphological defects in feline embryos, their competence to reach the blastocyst stage, and their ability to hatch. Oocyte-cumulus complexes were isolated after the scarification of ovaries and matured in vitro. Matured oocytes were fertilized in vitro by capacitated spermatozoa. Randomly selected oocytes were observed by TLM for seven-to-eight days. Out of 76 developed embryos, 41 were morphologically normal, of which 15 reached the blastocyst stage. Of 35 abnormally developed embryos, 17 reached the blastocyst stage, of which six had single aberrations and 11 had multiple aberrations. The hatching rate (%) was 15.6% in normally cleaving embryos, 6.25% in embryos with single aberrations, and 3.33% in those with multiple aberrations. The present study reports the first results, found by using TLM, about the frequency of the morphological defects of feline embryos, their competence to reach the blastocyst stage, and their ability to hatch.

Difficult embryo transfer has a negative impact on the outcome of in vitro fertilization

2003 ◽  
Vol 79 (3) ◽  
pp. 654-655 ◽  
Author(s):  
Steven D Spandorfer ◽  
Jessica Goldstein ◽  
José Navarro ◽  
Lucinda Veeck ◽  
Owen K Davis ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Transfer ◽  
Negative Impact ◽  
Vitro Fertilization

134 EFFECTS OF VITAMIN E AND VITAMIN C ON THE DEVELOPMENTAL COMPETENCE OF BUFFALO (BUBALUS BUBALIS) EMBRYOS DERIVED FROM PARTHENOGENETIC ACTIVATION, IN VITRO FERTILIZATION, AND NUCLEAR TRANSFER

2011 ◽  
Vol 23 (1) ◽  
pp. 171
Author(s):  
F. Lu ◽  
Z. Zhang ◽  
S. Zhang ◽  
N. Li ◽  
J. Jiang ◽  
...  
Keyword(s):  
Vitamin E ◽  
Nuclear Transfer ◽  
Bubalus Bubalis ◽  
Development Rate ◽  
The Other ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Vitro Fertilization ◽  
Parthenogenetic Activation

The purpose of this study was to explore the effects of vitamin E (VE) and vitamin C (VC) on the in vitro development of embryos derived from parthenogenetic activation (PA), in vitro fertilization (IVF), and somatic cell nuclear transfer (NT) in buffalo (Bubalus bubalis). Buffalo oocytes obtained from ovaries at slaughter were matured in vitro for 22 to 24 h. After maturation, oocytes were separated to 3 groups: one group of oocytes was fertilized in vitro with buffalo sperm; one group of oocytes was parthenogenetically activated by exposing them to 5 μM ionomycin for 5 min and then cultured in 2 mM 6-DMAP for 3 h; the other group of oocytes was enucleated, and fibroblasts in DMEM + 10% FBS for 4 to 5 days were transferred into enucleated oocytes by electronic fusion (100 v mm–1, 15 μs, and 3 pulses). After fusion, the activation of reconstructed embryos was induced by exposure to 5 μM ionomycin for 5 min and then cultured in 2 mM 6-DMAP for 3 h. The embryos of PA, IVF, and NT were respectively cultured in the culture medium (CM) containing different concentrations of VE, VC, or VE + VC for 7 to 9 days to evaluate embryonic development. As a result, when the embryos were cultured in the CM with different concentrations of VE (0, 50, 100, 150, and 200 μM), the blastocyst development rate of the embryos derived from PA, IVF, and NT gradually rose with increasing concentrations of VE and reached the highest amount [PA: 32.9% (81/246); IVF: 21.4% (45/210); and NT: 21.1% (47/223)] in the group containing 150 μM of VE; it was significantly higher than that of other groups (P < 0.05). When the different concentrations of VC (0, 50, 100, 150, and 200 μM) were added to the CM, the blastocyst development rate of the embryos derived from PA, IVF, and NT also enhanced according to the increasing concentration of VC, and more embryos developed to blastocysts in the group containing 150 μM of VC [PA: 31.2% (72/231); IVF: 20.2% (43/213); NT: 19.8% (48/243)] than in the other groups (P < 0.05). Compared with the control group (0 μM), the blastocyst rate of PA and IVF, as well as NT embryos, cultured in the CM with 150 μM VE + 150 μM VC groups was significantly higher (P < 0.05), but there were no significant differences in the percentage of blastocysts among groups of the 150 μM VE, 150 μM VC, and 150 μM VE + 150 μM VC (P > 0.05). These results indicated that adding VE (150 μM), VC (150 μM), or VE (150 μM) + VC (150 μM) in the CM could efficiently enhance the developmental competence of buffalo embryos during in vitro culture. This work was funded by China High Technology Development Program (2007AA100505), Guangxi Science Foundation (0718005-3A), Fok Ying Tung Education Foundation (111034).

205 EFFECT OF OSTEOPONTIN ON CLEAVAGE AND BLASTOCYST RATES IN BUFFALO SPECIES (BUBALUS BUBALIS)

2009 ◽  
Vol 21 (1) ◽  
pp. 200
Author(s):  
S. Di Francesco ◽  
E. Mariotti ◽  
M. Rubessa ◽  
G. Campanile ◽  
R. Di Palo ◽  
...  
Keyword(s):  
Embryo Development ◽  
Cumulus Cells ◽  
Bubalus Bubalis ◽  
The Other ◽  
Control Group ◽  
Single Chain ◽  
Cleavage Rate ◽  
Chi Square ◽  
Vitro Fertilization

It was previously reported that osteopontin (OPN), an acidic single-chain phosphorylated glycoprotein found in the oviductal fluid in cattle (Gabler C et al. 2003 Reproduction 126, 721–729), is able to facilitate fertilization in this species (Gasparrini B et al. 2008 Reprod. Fertil. Dev. 20(Suppl. I), 180 abst). The present study aimed to investigate whether the addition of OPN to the fertilization medium would affect both cleavage and postfertilization embryo development in the buffalo. To assess the influence of OPN on cleavage and blastocyst rates, in vitro-matured oocytes were fertilized in modified Tyrode’s albumin lactate pyruvate medium (Lu KH et al. 1987 Vet. Rec. 121, 259–260) supplemented with penicillamine, hypotaurine, and heparin, in the presence of 0.0 (n = 258), 0.1 (n = 263), 1 (n = 261), and 10 μg mL–1 (n = 264) of OPN. In vitro fertilization was carried out with frozen–thawed spermatozoa from a bull already tested for IVF. After 20 to 22 h of co-incubation at 38.5°C and 5% CO2 in air, putative zygotes were gently pipetted to remove cumulus cells, washed, and transferred, 10 per droplet, into 20 μL of SOF medium including essential and nonessential amino acids and BSA (Tervit HR et al. 1972 J. Reprod. Fertil. 30(3), 493–497), in a controlled gas atmosphere consisting of 5% CO2, 7% O2, and 88% N2, in humidified air, at 38.5°C. The culture medium was changed on Day 5 (Day 0 = day of insemination), when cleavage rate was assessed and embryos were moved into fresh medium for an additional 2 days. On Day 7, development rates into blastocysts of superior quality were recorded. Differences in the percentages of both cleavage and blastocyst rates among groups were analyzed by chi-square test. Significantly higher cleavage rates (59.3, 70.3, 71.6, and 42.4%, respectively, in the control group and in the groups with 0.1, 1, and 10 μg mL–1 of OPN; P < 0.01) were observed in the groups with 0.1 and 1 μg mL–1 of OPN compared with the other groups. Likewise, higher blastocyst rate percentages (17.4, 27.4, 29.9, and 9.5%, respectively, in the control group and in the groups with 0.1, 1, and 10 μg mL–1 of OPN; P < 0.01) were observed in the groups with 0.1 and 1 μg mL–1 of OPN compared with the other groups. In conclusion, these results showed that addition of low concentrations of OPN in the fertilization medium improved both cleavage and postfertilization embryo development in the buffalo, whereas the higher concentration resulted in impaired late-stage embryo development.

179 MODULATION OF RHOPHILIN-2 MAY REGULATE THE PROGRESSION OF CELL DIVISION IN FERTILIZED MOUSE EGGS

2008 ◽  
Vol 20 (1) ◽  
pp. 169
Author(s):  
T. Matsuoka ◽  
M. Tokoro ◽  
S. Shin ◽  
T. Amano ◽  
Y. Hosoi ◽  
...  
Keyword(s):  
Cell Division ◽  
Clostridium Botulinum ◽  
Cellular Localization ◽  
Nuclear Division ◽  
Effector Proteins ◽  
The Other ◽  
Vitro Fertilization ◽  
Gaba A ◽  
Mouse Eggs

It has been reported that activity of Rho, one of the GTPases, is essential for division of nuclei and cytoplasm of fertilized mouse eggs. Since it has been reported that alteration of activities of GTPases modifies their ability to attach to each of their effector proteins in somatic cells, effector proteins seem to be able to control not only progression but also repression of cell division by changing their cellular localizations through activities of GTPases. For this reason, Rhophilin-2, one of the effector proteins of Rho, seems to be involved in the decision of progression of division of fertilized mouse eggs. To examine whether this involvement works in fertilized mouse eggs, cellular localization of Rho and Rhophilin-2 in fertilized mouse eggs that were treated with Rho inhibitor were analyzed. Moreover, cellular localization of GABA A receptor association protein (GABARAP), which was identified in our previous study (Matsuoka et al. 2006 Reprod. Fertil. Dev. 18, 176–177) as a protein that interacts with Rhophilin-2, was also analyzed. Fertilized mouse eggs were obtained from in vitro fertilization technique. One group of fertilized eggs was obtained at 24 h after insemination as experimental control. To obtain the mouse eggs in which Rho activities were inhibited, Clostridium botulinum C3 exoenzyme (C3-CB), an inhibitor of Rho activity, was injected into the other group of fertilized mouse eggs at 12 h after insemination, and were collected after 12 h of subsequent culture. Cellular localization of Rho (n = 100), Rhophilin-2 (n = 10). and GABARAP (n = 10) in the collected oocytes was analyzed by using immunofluorescence. Our results showed that Rho and Rhophilin-2 were co-localized at the midbody microtubule, which is an important device for cytoplasmic division in control eggs. However, the inhibition of Rho activity did not modify the co-localization of Rho and Rhophilin-2. On the other hand, localization of GABARAP was modified by the inhibition of Rho activity, and GABARAP was detected around the nuclei of fertilized eggs in which Rho activity was inhibited. In the next experiment, we examined whether interaction of Rhophilin-2 and GABARAP was modified by the inhibition of Rho activity by using a co-immunoprecipitation assay (co-IP) (n = 100). The interaction of Rhophilin-2 and GABARAP was found to disappear after inhibition of Rho activity. These results suggest that activity of Rho seems to regulate cytoplasmic division through Rhophilin-2 modification. Moreover, Rho seem to modulate the nuclear division of fertilized mouse eggs by regulating the interaction between Rhophilin-2 and GABARAP. This study was supported by a Grant-in-Aid for the 21st Century COE Program of the Japan Mext and by a grant for the Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technology Excellence of the JST.

52 USE OF TRANSWELL CELL CULTURE AND 3-DIMENSIONAL PRINTING TECHNOLOGY TO DEVELOP AN IN VITRO BOVINE OVIDUCT

2016 ◽  
Vol 28 (2) ◽  
pp. 156 ◽  
Author(s):  
M. A. M. M. Ferraz ◽  
H. H. W. Henning ◽  
K. M. A. Van Dorenmalen ◽  
P. L. A. M. Vos ◽  
T. A. E. Stout ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Growth ◽  
Liquid Culture ◽  
Primary Cilia ◽  
Negative Impact ◽  
Perfusion Culture ◽  
Vitro Fertilization ◽  
Custom Made

Oviduct epithelial cells (OECs) generate the microenvironment for mammalian fertilization. When cultured in vitro OECs rapidly lose their differentiated cell properties (e.g. secretory activity and cilia), while suspended cells have a limited lifespan. These limitations, likely due to the lack of folded tubular geometry of the oviduct, prompted us to combine transwell cell culture and 3-D printing technologies to mimic the in vivo OEC niche in order to better study the unique role of the oviduct and its microenvironment during the processes of fertilization and early embryonic development. U-shape inserts were 3-D printed using a multi-arm acrylate-based resin (PIC100) on an Envisiontec Perfactory P3 stereolithographer. Post-printing treatments of custom-made tubular transwell inserts were first tested in order to determine any possible negative impact of the plastics on cell growth. Inserts were either untreated, post-cured with 1000 flashes/side (Otoflash, 66 W), post-cured and Soxhlet-extracted overnight in isopropanol, or post-cured and Soxhlet-extracted over the weekend in water at 37°C. The post-cured and Soxhlet-extracted overnight in isopropanol inserts were selected as best pretreatment for culturing OECs. These inserts were mounted with track-etched PET membranes (12 µm thick, 0.4 µm pore diameter) to create a U-shape geometry that allows perfusion. Bovine OECs were obtained by squeezing the whole oviduct collected from slaughterhouse cows (on luteal phase) and cultured as monolayers for 7 days (n = 2 cows). These de-differentiated OECs were seeded on the membranes, grown to confluence (7 days), and cultured (1) at an air-liquid interface for 6 and 14 days (air-liquid culture) or (2) under perfusion (6 mL h–1) for 6 days (perfusion culture). OECs were also cultured on coverslips as monolayers (2-D culture) for 6 and 14 days. After this period, the OECs were fixed and immune labelled to determine their polarized state. Polarization of OECs (laminin and primary cilia detection) was observed on Day 6 for perfusion culture, on Day 14 for air-liquid culture, and was not detected in 2-D culture. The presence of secondary cilia (acetylated α-tubulin) was observed in 6% of the cells cultured under perfusion at Day 6; secondary cilia was not present in air-liquid or 2-D cultures during the period analysed. In conclusion, post-curing and Soxhlet extraction of leachable compounds is crucial to avoid toxic effects on cell growth. The U-shape custom-designed inserts are able to create a tube-like surface in which bovine oviducal cells can be cultured to confluency and thereafter repolarize (presence of primary cilia and detection of laminin); this polarization occurs faster when the U-shape culture is under perfusion. Further studies will examine the ability of the cells to differentiate further (development of secondary cilia and secretory ability) and support in vitro fertilization. To this end, 3-D designs will be tested to determine their use for live cell imaging and for collecting secreted fluids.

P–007 Effect of swim-up on sperm morphology

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
R Ganeva ◽  
D Parvanov ◽  
M Handzhiyska ◽  
G Stamenov
Keyword(s):  
Sperm Morphology ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Lower Percentage ◽  
Preparation Technique ◽  
Sperm Preparation ◽  
Multiple Defects ◽  
Before And After ◽  
Registration Number ◽  
Morphological Defects

Abstract Study question To evaluate the effect of swim-up on the percentage of certain morphological defects in the semen population Summary answer Swim-up preparation led to significantly lower percentage of spermatozoa with cytoplasmic droplets, thick neck and also multiple defects. What is known already Swim-up is routinely used sperm preparation technique in ART practice. It is widely known that swim-up enhances sperm quality in terms of motility and sperm morphology. However, the effect of swim-up on the frequency of occurrence of the specific sperm morphological abnormalities is still missing. Study design, size, duration This observational study involved 30 teratozoospermic patients of Nadezhda Women’s Health hospital between December 2020 and January 2021. Sperm morphology was evaluated before and after swim-up preparation. Participants/materials, setting, methods Native semen was liquefied and was subjected to swim up. Semen analysis performed according to WHO 2010. Native semen and swim up samples from the same men were subjected to Kruger strict morphological evaluation. The analyzed sperm morphological defects included: head defects (large, small, tapered, pyriform, round, amorphous and double heads); midpiece defects (bent, asymmetrical, thin, thick, presence of cytoplasmic droplet); tail defects (short, hairpin, bent, coiled tail and terminal droplet) and multiple defects. Main results and the role of chance Wilcoxon paired test showed that the percentage of morphologically normal spermatozoa was significantly higher in the swim-up samples in comparison to the native semen (8.5±4.2% vs 4.9±3.2%, p &lt; 0.05). In addition, the percentage of spermatozoa bearing multiple defects was found to be significantly lower in the swim-up samples than in the native semen (25.8±11.6% vs 37.0±15.0%, p &lt; 0.05). Two specific sperm morphological defects were found to be significantly lower after swim-up preparation: the presence of cytoplasmic droplets (6.0±1.0% vs 8.6±1.5%, p &lt; 0.05) and the thick neck (9.7±5.5% vs 12.8±5.8%, p &lt; 0.05). No significant different were observed in the other morphological defects between swim up samples and native semen (p &gt; 0.05). Limitations, reasons for caution Results obtained from this study need to be confirmed by larger group of samples. Wider implications of the findings: Our study showed a significant reduction of certain midpiece defects after swim-up. The observed selection of spermatozoa without thick necks and cytoplasmic droplets explains the effectiveness of swim-up on ART. In addition, the obtained results can serve as a guide for future validation of new sperm preparation techniques. Trial registration number Not applicable

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