Separation of T and B lymphocytes from human peripheral blood mononuclear cells using density perturbation methods.
The fractionation of sub-populations of peripheral blood mononuclear cells has become an essential routine procedure and some of the main fractionation methods used today are immunomagnetic separations. We describe a less expensive method for the separation of subpopulations of mononuclear cells using density perturbation, which uses the binding of antibody-coated dense polystyrene beads to increase the density of specific sub-populations of cells. By incubating a total mononuclear fraction from human peripheral blood together with antibody-coated beads, in a commercially-available lymphocyte separation medium (Nycoprep 1.077), a depletion of 94.9 +/- 1.68% of the T cells could be obtained by this procedure; a depletion of 69.7 +/- 1.78% of the B cells was also achieved. These results indicate the potential for the separation of different sub-populations of peripheral blood mononuclear cells on the basis of the immunological identity of the surface of cells using density perturbation methods involving antibody-coated dense polystyrene beads.
Department of Biology, University of Essex, Wivenhoe Park, Colchester, UK.
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Patel
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Rubbi
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Rickwood
D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000502:physiology*; D002469:Cell Separation; D003470:Culture Media; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008863:Microspheres; D009000:Monocytes / Q000502:physiology*; D013601:T-Lymphocytes / Q000502:physiology*
D000911:Antibodies, Monoclonal; D003470:Culture Media