Fractionation of cells and subcellular particles with Percoll

At present, centrifugation is the most common method for separation and isolation of cells and subcellular particles. The technique can be used for a wide range of applications. During latter years it has become obvious what a powerful method density gradient centrifugation is, especially when used...

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Bibliographic Details
Published in:Journal of Biochemical and Biophysical Methods 2000-07, Vol.44 (1), p.1-30
Main Author: Pertoft, Håkan
Format: Article
Language:English
Subjects:
Animals
Cattle
Cell Fractionation - methods
Cell Separation - methods
Cells
Centrifugation
Centrifugation, Density Gradient - instrumentation
Centrifugation, Density Gradient - methods
Density gradients
Humans
Male
Osmolar Concentration
Percoll
Povidone - chemistry
Separation
Silicon Dioxide - chemistry
Spermatozoa - ultrastructure
Subcellular particles
Sucrose
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Summary:At present, centrifugation is the most common method for separation and isolation of cells and subcellular particles. The technique can be used for a wide range of applications. During latter years it has become obvious what a powerful method density gradient centrifugation is, especially when used in conjunction with sensitive assays or clinical treatments. The most active areas for use of density gradient centrifugation include purification for in vitro fertilization of sperm of both human and bovine origin, isolation of cells for cell therapy of patients receiving chemo- and radiation therapy and basic research both on cellular and subcellular levels. These treatments and investigations require homogeneous populations of cells and cell organelles, which are undamaged after the separation procedure. Percoll, once introduced to reduce convection during centrifugation, has proved to be the density gradient medium of choice since it fulfills almost all criteria of an ideal density gradient medium. Recently good results have also been obtained after silanization of colloidal silica particles, e.g. BactXtractor™. The latter medium has proved to be useful in recovery of microorganisms from food samples free of inhibitors to the Polymer Chain Reaction (PCR). The separation procedures described for Percoll in this review seem to be applicable to any cells or organelles in suspension for which differences in size or bouyant density exist. Furthermore, since Percoll media are inert, they are well suited for the separation of fragile elements like enveloped viruses.
ISSN:0165-022X
1872-857X
DOI:10.1016/S0165-022X(00)00066-X