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%.

Separation of Isospora (Toxoplasma) gondii cysts and cystozoites from mouse brain tissue by continuous density - gradient centrifugation

Parasitology ◽  
1981 ◽  
Vol 83 (1) ◽  
pp. 103-108 ◽  
Author(s):  
A. W. C. A. Cornelissen ◽  
J. P. Overdulve ◽  
J. M. Hoenderboom
Keyword(s):  
Toxoplasma Gondii ◽  
Brain Tissue ◽  
Density Gradient ◽  
Infected Mouse ◽  
Mouse Brain ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Percoll Gradient ◽  
Isolation And Purification ◽  
Reproducible Method

SUMMARYA simple, quick and reproducible method consisting of density-gradient centrifugation of homogenized infected mouse brain tissue on Percoll is described for the isolation and purification of cysts of Isospora (Toxoplasma) gondii. A 100% recovery of cysts, with 74·2% in a single fraction with a specific gravity of 1·056, was obtained by overlaying homogenates of infected mouse brains on a pre-formed Percoll gradient and centrifugation at low g forces. With this procedure recovery was independent of the age of the cysts. Titration of purified cystozoites showed there to be no loss of infectivity.

Separation and characterization of Leydig cells and macrophages from rat testes

1991 ◽  
Vol 130 (3) ◽  
pp. 357-365 ◽  
Author(s):  
G. Dirami ◽  
L. W. Poulter ◽  
B. A. Cooke
Keyword(s):  
Monoclonal Antibody ◽  
Density Gradient ◽  
Leydig Cells ◽  
Magnetic Beads ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Sedimentation Velocity ◽  
Average Density ◽  
Centrifugal Elutriation ◽  
Percoll Gradients

ABSTRACT A method involving centrifugal elutriation followed by density gradient centrifugation and incubation with a macrophage monoclonal antibody has been investigated to separate and characterize Leydig cells and macrophages from adult rat testes. After dispersion of the testes with collagenase, the isolated interstitial cells were found to contain 18% Leydig cells and 12% macrophages. These cells were then separated by centrifugal elutriation into eight fractions (F1–F8) (9 to 74 ml/min at 386 g). Each of these fractions was then further purified by density gradient centrifugation on 0–90% Percoll gradients. After centrifugal elutriation, the macrophages were mainly eluted in the first three fractions (F1–F3), whereas the Leydig cell percentage increased in each fraction with increasing flow rate. After further purification of each fraction on Percoll gradients, high percentages of macrophages (11–20%) were found in fractions F1–F3 (average density 1·045 g/ml), containing 11–37% Leydig cells. Less than 3% of the cells in fraction F4–F8 (average density 1 ·075 g/ml) were macrophages and more than 95% were Leydig cells. Heterogeneity of Leydig cells with respect to sedimentation velocities and function was found. Leydig cells from elutriated-and Percoll-purified fractions F4–F8 were heterogeneous with respect to testosterone and cyclic AMP (cAMP) production but showed a similar binding capacity for 125I-labelled human chorionic gonadotrophin. Leydig cells with the highest sedimentation velocity (35·7 mm/h-g) from fractions F7 and F8 were approximately twofold more responsive to LH (3·3 nmol/l) with respect to testosterone and cAMP production compared with Leydig cells with the lowest sedimentation velocity (20·7 mm/h-g). The elutriated and Percoll-purified cells (corresponding to fractions F4–F8) were further purified by incubation with magnetic beads coated with a macrophage monoclonal antibody; this yielded very pure Leydig cells containing <0·3% macrophages. The incubation temperature (room temperature or 4 °C) during the purification with magnetic beads did not affect the degree of purity or the responsiveness of the Leydig cells to LH. The removal of the remaining macrophages with magnetic beads did not have any significant effect on the Leydig cell responsiveness to LH. It was concluded that Leydig cells purified by elutriation and density gradient centrifugation are heterogeneous with respect to their sedimentation velocities and responses to LH; the higher the sedimentation velocity, the higher is their capacity to respond to LH. Leydig cells free from macrophages can be prepared by further purification using magnetic beads coated with a macrophage monoclonal antibody. Journal of Endocrinology (1991) 130, 357–365

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

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.

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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