Separation of viable, motile sperm from red blood cells and dead spermatozoa: A comparison of four density gradient centrifugation media in the dog

2008 ◽  
Vol 70 (3) ◽  
pp. 578-579
Author(s):  
T.C. Phillips ◽  
G. Dhaliwal ◽  
J.P. Verstegen
Keyword(s):  
Red Blood Cells ◽  
Density Gradient ◽  
Blood Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Motile Sperm

scholarly journals Isolation of blood cells of Ascidiacea using density gradient centrifugation.

Dobutsu seiri ◽  
1988 ◽  
Vol 5 (4) ◽  
pp. 177-181
Author(s):  
Hitoshi MICHIBATA ◽  
Taro UYAMA ◽  
Yukiya IWATA ◽  
Junko HIRATA
Keyword(s):  
Density Gradient ◽  
Blood Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation

P–019 Sperm parameter and ICSI / IVF outcomes after sperm selection using microfluidic sperm separator and density gradient centrifugation with swim-up in split semen sample

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
R Higashiyama ◽  
M Kishimoto ◽  
S Komure ◽  
S Mizuta ◽  
K Kitaya ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Sperm Concentration ◽  
Development Rate ◽  
Sperm Dna Fragmentation ◽  
Motile Sperm ◽  
Fragmentation Rate ◽  
Sperm Dna

Abstract Study question To analyze whether microfluidic sperm selection (MSS) by ZyMōt™ improves sperm DNA fragmentation rate and embryonic development compared to density gradient centrifugation with swim-up (DGCS). Summary answer MSS by ZyMōt™ selects sperm for clinical use with less DNA damage significantly compared to DGCS. What is known already Conventional sperm preparation methods, such as density gradient centrifugation and the swim-up method utilize centrifugation during processing, may damage the sperm. MSS may allow for improved selection of normal sperm compared with conventional sperm preparation as it yields sperm with a lower DNA fragmentation rate. However, there are few clinical studies by sibling oocytes study compared to DGCS. Study design, size, duration This prospective study was performed between March 2020 and May 2020 at a reproductive center. All patients involved gave written consent, and institutional review board approval was granted. A total of 575 metaphase II oocytes were collected from 49 cycles. Wife’s age was 34.7 ± 3.9 years old. Raw sperm concentration and motile sperm concentration was 63.1 ± 78.7M/mL, and 41.6 ± 67.7M/mL, respectively. Participants/materials, setting, methods Patients who performed ART for the first or second time were divided into two groups according to MSS and DGCS. Sperm DNA fragmentation rate (SDFR) and motile sperm concentration were compered between MSS and DGCS. SDFR was measured by sperm chromatin structure assay (SCSA) using a flow cytometer. Sibling oocytes were randomized into MSS-IVF, DGCS-IVF, MSS-ICSI, and DGCS-ICSI. Rate of two pronuclear (2PN) oocytes, blastocysts development, and good-quality blastocysts were compared between each group. Main results and the role of chance SDFR was 13.5 ± 11.8% for raw semen. SDFR was significantly lower after MSS (3.6 ± 4.1%) than that for raw semen and after DGCS (17.4 ± 14.8%) (P < 0.01). Motile sperm concentration after MSS (19.0 ± 28.3M/mL) was significantly higher after than after DGCS (15.4 ± 15.3M/mL) (P < 0.01). The number of IVF performed was 145 for MSS and 132 for DGCS. IVF results (MSS vs DGCS) were 2PN rate (73.1% vs 72.0%), blastocysts development rate (65.3% vs 55.4%), and good quality blastocysts rate (43.2% vs 34.9%). The number of ICSI performed was 149 for MSS and 149 for DGCS. ICSI results (MSS vs DGCS) were 2PN rate (77.9% vs 79.2%), blastocysts development rate (68.8% vs 65.8%), and good quality blastocysts rate (35.8% vs 30.6%). No significant difference was observed between MSS and DGCS for each parameter both IVF and ICSI. Limitations, reasons for caution The participants were limited to those who collected semen of 2mL or more and motile sperm concentration of above 1M/mL, because semen sample needed to be divided to MSS and DGCS. Wider implications of the findings: This is the first study to conducted in sibling oosytes study with MSS and DGCS, in both IVF and ICSI. MSS is effective in collecting sperm with less DNA damage compared to DGCS. Motile sperm concentration after using MSS is sufficient to perform IVF as well as DGCS. Trial registration number Not applicable

scholarly journals An automated instrument for intrauterine insemination sperm preparation

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Alex Jafek ◽  
Haidong Feng ◽  
Hayden Brady ◽  
Kevin Petersen ◽  
Marzieh Chaharlang ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Blood Cells ◽  
Density Gradient Centrifugation ◽  
Intrauterine Insemination ◽  
Gradient Centrifugation ◽  
White Blood Cells ◽  
Operating Time ◽  
Motile Sperm ◽  
Sperm Preparation ◽  
Automated Instrument

AbstractSperm preparation is critical to achieving a successful intrauterine insemination and requires the processing of a semen sample to remove white blood cells, wash away seminal plasma, and reduce sample volume. We present an automated instrument capable of performing a sperm preparation starting with a diluted semen sample. We compare our device against a density gradient centrifugation by processing 0.5 mL portions of patient samples through each treatment. In 5 min of operating time, the instrument recovers an average of 86% of all sperm and 82% of progressively motile sperm from the original sample while removing white blood cells, replacing the seminal plasma, and reducing the volume of the sample to the clinically required level. In 25 min of operating time, density gradient centrifugation recovers an average of 33% of all sperm and 41% of progressively motile sperm. The automated instrument could improve access to IUI as a treatment option by allowing satellite doctor’s offices to offer intrauterine insemination as an option for patients without the clinical support required by existing methods.

scholarly journals Separation and Optimisation of a Sucrose Density Gradient Centrifugation Protocol for Isolation of Peripheral Blood Mononuclear Cells (PBMC)

2018 ◽  
pp. 1-4
Author(s):  
Sudeep Nagaraj ◽  
Shubha Nivargi ◽  
Leelavathy Nanjappa ◽  
Jagadish Tavarekere Venkataravanappa
Keyword(s):  
Density Gradient ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Mononuclear Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
One Step

One step centrifugation procedure used commonly for separation of blood cells is the ficoll gradient centrifugation. In this method, after centrifugation, the peripheral blood mononuclear cells (PBMCs) are located on the top of the separation fluid, whereas other blood cells erythrocytes and granulocytes sediment to the bottom. In the present study 75% of lymphocyte suspension could be separated by using a one-step density gradient centrifugation of sodium heparin blood with Sucrose. Sucrose was diluted into different concentrations using miliQ water (10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 100%,). 4 mL of diluted blood was layered on 4 mL of each sucrose solution and centrifuged for 45 minutes at 1000 rpm. Clear separation of PBMCs could be observed in solution with 40% sucrose. The separated PBMCs were analysed in haeme analyser which showed 75% lymphocytes, 23% monocytes and 2% of other cells.

The Nature of Monkey Cell Haemagglutinins of Rosen’s Group II Adenoviruses

1965 ◽  
Vol 20 (6) ◽  
pp. 560-563 ◽  
Author(s):  
R. Wigand ◽  
M. Stöhr
Keyword(s):  
Density Gradient ◽  
Blood Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Buoyant Density ◽  
Haemagglutination Inhibition ◽  
Wild Strain ◽  
Rat Blood ◽  
Monkey Cell ◽  
Group Ii

Some of the adenovirus type of Rosen’s group II exhibit weak agglutinating properties for monkey blood cells. The relation of these haemagglutinins to haemagglutinins for rat blood cells and to the infectious virus was studied by cross-absorption with blood cells and by density gradient centrifugation with caesium chloride. In the virus types studied (type 9, 13, and a wild strain of type 15) one part of the monkey cell haemagglutinin was associated with the virion and, for type 9 and 15, was inseparable from the virus-associated haemagglutinin for rat blood cells. In density gradient centrifugation a second haemagglutinin for monkey blood cells was found, separable from the virus particles and associated with the portion of higher buoyant density of the soluble haemagglutinin for rat blood cells. In type 13 and 23, haemagglutinins for monkey and rat blood cells can be separated by absorption with the heterologous blood cell species.Haemagglutinins for monkey cells are resistant to trypsin or trypsin plus subsequent heating to 55°C, in contrast to haemagglutinins for rat blood cells. Monkey cell haemagglutination is reversibly inhibited by the presence of CsCl.In haemagglutination-inhibition tests it was found that haemagglutination with monkey blood cells is more difficult to be inhibited by immune serum that haemagglutination with rat blood cells.

Separation of viable microfilariae free of blood cells on Percoll gradients

1984 ◽  
Vol 58 (1) ◽  
pp. 69-70 ◽  
Author(s):  
R. Chandrashekar ◽  
U. R. Rao ◽  
G. R. Rajasekariah ◽  
D. Subrahmanyam
Keyword(s):  
Density Gradient ◽  
Blood Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Brugia Pahangi ◽  
Percoll Gradients ◽  
Reproducible Method ◽  
Dipetalonema Viteae

AbstractA consistent and reproducible method is described for isolating pure populations of microfilariae of Litomosoides carinii, Brugia pahangi, B. malayi and Dipetalonema viteae, free of cells, from blood, by density gradient centrifugation on Percoll in 0.25M sucrose. The recovery of the microfilariae was 85 to 97%.

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.

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