Characterization of human mononuclear cells after positive selection with immunomagnetic particles.
We have investigated the possibility to employing magnetic monodisperse polymer particles for positive selection of human peripheral blood mononuclear cell populations. By carefully titrating the ratio between particles and cells we succeeded in isolating a number of cell populations that could be cultivated subsequently in vitro for functional studies. The success of the procedure is partly dependent on the properties of the monoclonal antibodies used to sensitize the cells. Provided these antibodies do not react with membrane structures involved in the transduction of activating signals, highly purified, quiescent cell populations can be recovered in a single fractionation step. In most instances particles will detach from the isolated cells by overnight culture, and the particles can then be removed from the system by a suitable magnet. T lymphocytes, subpopulations of T lymphocytes, and B lymphocytes have been isolated in this way and studied in a variety of functional assay systems. Comparison with cells obtained after negative selection clearly demonstrates the usefulness of this technique, especially if the membrane marker selected for it is not directly engaged in the activation processes.
T
T
Lea
E
E
Smeland
S
S
Funderud
F
F
Vartdal
C
C
Davies
K
K
Beiske
J
J
Ugelstad
D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000145:classification / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D006801:Humans; D007148:Immunoglobulin mu-Chains / Q000276:immunology; D007700:Kinetics; D008213:Lymphocyte Activation; D008280:Magnetics; D012313:RNA / Q000096:biosynthesis; D011947:Receptors, Antigen, B-Cell / Q000276:immunology; D013601:T-Lymphocytes / Q000145:classification / Q000166:cytology* / Q000276:immunology
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D007148:Immunoglobulin mu-Chains; D011947:Receptors, Antigen, B-Cell; D012313:RNA