Effects of density-gradient centrifugation and modified swim-up methods on stallion sperm quality

2016 ◽  
Vol 43 ◽  
pp. S58
Author(s):  
S.R. Teague ◽  
S.R. Lambourne ◽  
K. Hinrichs ◽  
Z. Gibb ◽  
C.C. Love ◽  
...  
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Effects Of Density

scholarly journals Improvement of Sperm Quality in Hyperviscous Semen following DNase I Treatment

2019 ◽  
Vol 2019 ◽  
pp. 1-8
Author(s):  
Effrosyni Nosi ◽  
Angelos D. Gritzapis ◽  
Konstantinos Makarounis ◽  
Georgios Georgoulias ◽  
Vasilios Kapetanios ◽  
...  
Keyword(s):  
Density Gradient ◽  
Sperm Motility ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Sperm Count ◽  
Gradient Centrifugation ◽  
Dnase I ◽  
Control Group ◽  
First Case ◽  
Dnase Treatment

Semen hyperviscosity impairs sperm motility and can lead to male infertility. This prospective study aimed at assessing the ability of exogenous DNase in improving sperm quality, taking into consideration that DNase has been found in the seminal plasma of several species and that neutrophils release chromatin in order to trap bacteria. A total of seventy-seven semen samples with high seminal viscosity (HSV) as the study group and sixty-two semen samples with normal seminal viscosity (NSV) as the control group were compared in this analysis. These semen samples were divided into three groups of receiving treatment (a) with DNase I at 37°C for 15 min, (b) by density gradient centrifugation, and (c) with a combination of the above two methods. Following a fifteen-minute treatment of hyperviscous semen, the motility of spermatozoa in 83% of semen samples increased to a statistically significant degree. On the contrary, DNase treatment of semen with normal viscosity had no such effects. The above treatment was also accompanied by a significant increase in the percentage of normal spermatozoa, resulting in a major decrease of the teratozoospermia index. Comparison between semen samples that underwent density gradient centrifugation following DNase I treatment, to those collected after density gradient treatment alone, showed that in the first case the results were more spectacular. The evaluation of each preparation in terms of yield (% total progressively motile sperm count after treatment in relation to the initial total sperm count) revealed that the combined approach resulted in 29.8% vs. 18.5% with density treatment alone (p=0.0121). DNase I treatment results in an improvement of sperm motility and morphology and could be beneficial to men with hyperviscous semen in assisted reproduction protocols.

scholarly journals Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial

2014 ◽  
Vol 12 (1) ◽  
pp. 121 ◽  
Author(s):  
Helena Malvezzi ◽  
Rakesh Sharma ◽  
Ashok Agarwal ◽  
Adel M Abuzenadah ◽  
Muhammad Abu-Elmagd
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Controlled Trial ◽  
Gradient Centrifugation

7 EFFECTS OF DIFFERENT TRYPSIN SOURCES AND CONCENTRATIONS ON THE VIABILITY OF BOVINE SPERM PRE- AND POST-CRYOPRESERVATION

2007 ◽  
Vol 19 (1) ◽  
pp. 121
Author(s):  
B. A. Blevins ◽  
S. Steenson ◽  
N. M. Loskutoff
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Trypsin Treatment ◽  
Porcine Pancreas ◽  
Bovine Semen ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Percoll Density Gradient Centrifugation

The goal of this research was to investigate the effect of different sources and concentrations of trypsin on the viability of bovine sperm as a potential method for removing pathogens similar to the washing methods developed for embryos. Trypsin derived from porcine pancreas (Sigma-Aldrich, St Louis, MO, USA) at 2.5% and 0.25% was compared to the recombinant human sequence (TrypLE Select; Invitrogen, Carlsbad, CA, USA) at 10× and 1× concentrations. Cryopreserved bovine sperm (n = 3 bulls) were thawed and processed using discontinuous (90/45) Percoll density gradient centrifugation; the sperm pellets were then washed (10 min at 300g) in TL-HEPES Solution (Cambrex Corp., East Rutherford, NJ, USA) and then resuspended in 1 mL of the same medium. Aliquots of 200 µL of the washed sperm were then added to 1 mL of each of the 4 trypsin treatments as well as a negative control (without trypsin) and incubated at room temperature. Aliquots (25 µL) of each treatment were examined for progressive motility after 5 min. As a result, the control sperm (no trypsin) increased progressive motility by 6.7% and the 1× TrypLE treatment by 9.3%. However, the 10× TrypLE Select and the 10× and 1× porcine pancreas extracts decreased progressive motility by 8.3, 29.0, and 4.0%, respectively. The objective of the second experiment was to determine if the treatment of bovine semen with trypsin (1× TrypLE Select and 0.25% porcine pancreas extract) before or after cryopreservation would affect sperm quality as compared to cryopreservation without trypsin treatment. Raw semen (n = 6 bulls) was collected, evaluated, cryopreserved, and then thawed using a standard bovine method (Biladyl®; Minitube, Verona, WI, USA) without further treatment (control) or after treatment with one of two trypsin treatments (density gradient centrifugation with 1× TrypLE Select or 0.25% porcine pancreas extract in the 45% Percoll layer and a soybean trypsin in activator (Sigma) in the 90% layer) either before freezing (Treat–Freeze) or after thawing (Freeze–Treat). The results for the 6 individual bull samples were comparable and are presented as means (± SEM) compared to the cryopreserved control (no trypsin treatment). Using the Mann–Whitney Rank Sum Test, no differences (P > 0.05) were found in any of the parameters comparing the crypreserved controls (no trypsin treatment) and the 4 treatments: Freeze–Treat vs. Treat–Freeze using either the recombinant TrypLE Select (1×) or the porcine pancreas extract (0.25%). These results suggest that trypsin treatment, before or after cryopreservation, can be used safely on bovine sperm without affecting viability in vitro. Table 1.Comparison of cryopreserved bovine semen without and with various treatments

scholarly journals Effect of Osmolarity and Density of Colloid Formulations on the Outcome of SLC-Selection of Stallion Spermatozoa

2011 ◽  
Vol 2011 ◽  
pp. 1-5 ◽  
Author(s):  
J. M. Morrell ◽  
A. Johannisson ◽  
H. Rodriguez-Martinez
Keyword(s):  
Density Gradient ◽  
Assisted Reproduction ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Single Layer ◽  
Normal Density ◽  
High Osmolarity ◽  
Sperm Survival ◽  
Selection Of

The osmolarity and density of colloids used to prepare spermatozoa for assisted reproduction may affect sperm quality in the resultant preparation. In this study, two osmolarities of Androcoll-E for single-layer or density gradient centrifugation of stallion spermatozoa were compared: “normal” (320 mOsm) or “high” (345 mOsm). Mean yields for the two centrifugation techniques did not differ between treatments or osmolarities (single layer centrifugation: 30.19±16.9×106 and 25.8±18.5×106 spermatozoa; density gradient centrifugation: 31.84±19.7×106 and 26.46±20.0×106 spermatozoa respectively for the two osmolarities). However, use of the high osmolarity colloid for single layer centrifugation increased the proportion of morphologically normal spermatozoa (P<.05). Therefore, increasing the osmolarity of the colloid formulation may be beneficial for processing ejaculates containing a high proportion of abnormal spermatozoa by SLC. Reducing the density of the colloid used for the SLC substantially increased the yield of motile spermatozoa compared to the normal density colloid (mean ± SD: 72.6±28.9×106 versus 28.9±24.7×106), while also prolonging sperm survival by 24 hours compared to the uncentrifuged ejaculate. This increased yield may render Single Layer Centrifugation practical for use in the field.

scholarly journals The Effects of In Vitro Incubation of Asthenoteratozoospermic Semen after Density Gradient Centrifugation at Room Temperature and 37C on Sperm Parameters, Chromatin Quality and DNA Fragmentation in a Short Time Period

2020 ◽  
Author(s):  
Motahareh Karimi Zarchi ◽  
Behnam Maleki ◽  
Mahmood Dehghani Ashkezari ◽  
Leila Motamed Zadeh ◽  
Azam Agha-Rahimi
Keyword(s):  
Dna Fragmentation ◽  
Density Gradient ◽  
Room Temperature ◽  
Density Gradient Centrifugation ◽  
Sperm Quality ◽  
Gradient Centrifugation ◽  
Sperm Parameters ◽  
Sperm Dna Fragmentation ◽  
In Vitro Incubation

Background: Sperm quality is an important factor in assisted reproductive technology (ART) that affects the success rate of infertile couples treatment. In vitro incubation of sperm can influence its parameters and DNA integrity. The present study focused on the effect of different incubation temperatures sperm parameters on asthenoteratozoospermia semen prepared with density gradient centrifugation at different times. Methods: Twenty-seven samples were collected and prepared. Then, the suspension was divided into two parts. One part was incubated at room temperature (RT), and another was incubated at 37°C. Immediately and after 2 hr (2H) and 4 hr (4H), sper-matozoa were evaluated regarding motility, viability, morphology, sperm protamine deficiency, chromatin and DNA fragmentation. Statistical analysis was performed using paired t-test and repeated measures. The p<0.05 was considered statistically significant. Results: Our results showed that following 2 and 4 hr of incubation at RT, sperm progressive motility and viability decreased significantly. Sperm DNA fragmentation increased significantly following 2 and 4 hr of incubation at RT and 37°C. The Trend analysis confirmed that there were no significant differences between sperm parameters and DNA fragmentation after different times at RT and 37°C. Conclusion: Incubation of sperm at RT in comparison to 37°C didn’t preserve sperm parameters and DNA efficiently. Therefore, IVF, ICSI and IUI procedure should be performed in the soonest possible time after sperm preparation.

Uranyl Acetate and Rotary Shadowing to Increase the Contrast of Small Aggregated Virus Particles

1969 ◽  
Vol 27 ◽  
pp. 424-425
Author(s):  
Lee F. Ellis ◽  
Richard M. Van Frank ◽  
Walter J. Kleinschmidt
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Uranyl Acetate ◽  
Interferon Inducer ◽  
Virus Particles ◽  
Staining Methods ◽  
Rotary Shadowing

The extract from Penicillum stoliniferum, known as statolon, has been purified by density gradient centrifugation. These centrifuge fractions contained virus particles that are an interferon inducer in mice or in tissue culture. Highly purified preparations of these particles are difficult to enumerate by electron microscopy because of aggregation. Therefore a study of staining methods was undertaken.

The Combining Ability of Glycoproteins IIb, IIIa and Ca2+ in EDTA-Treated Nonaggregable Platelets

1983 ◽  
Vol 50 (04) ◽  
pp. 848-851 ◽  
Author(s):  
Marjorie B Zucker ◽  
David Varon ◽  
Nicholas C Masiello ◽  
Simon Karpatkin
Keyword(s):  
Density Gradient ◽  
Combining Ability ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Electrophoretic Pattern ◽  
Sucrose Density Gradient ◽  
Irreversible Loss ◽  
Sucrose Density Gradient Centrifugation ◽  
Crossed Immunoelectrophoresis ◽  
Triton X

SummaryPlatelets deprived of calcium and incubated at 37° C for 10 min lose their ability to bind fibrinogen or aggregate with ADP when adequate concentrations of calcium are restored. Since the calcium complex of glycoproteins (GP) IIb and IIIa is the presumed receptor for fibrinogen, it seemed appropriate to examine the behavior of these glycoproteins in incubated non-aggregable platelets. No differences were noted in the electrophoretic pattern of nonaggregable EDTA-treated and aggregable control CaEDTA-treated platelets when SDS gels of Triton X- 114 fractions were stained with silver. GP IIb and IIIa were extracted from either nonaggregable EDTA-treated platelets or aggregable control platelets with calcium-Tris-Triton buffer and subjected to sucrose density gradient centrifugation or crossed immunoelectrophoresis. With both types of platelets, these glycoproteins formed a complex in the presence of calcium. If the glycoproteins were extracted with EDTA-Tris-Triton buffer, or if Triton-solubilized platelet membranes were incubated with EGTA at 37° C for 30 min, GP IIb and IIIa were unable to form a complex in the presence of calcium. We conclude that inability of extracted GP IIb and IIIa to combine in the presence of calcium is not responsible for the irreversible loss of aggregability that occurs when whole platelets are incubated with EDTA at 37° C.

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