CD20 positive human B lymphocytes separated with the magnetic cell sorter (MACS) can be induced to proliferation and antibody secretion in vitro.
A method is described for the efficient purification of human B lymphocytes from peripheral blood by magnetic separation. Biotinylated, superparamagnetic particles were coupled to target cells by fluorescein isothiocyanate conjugated avidin and biotinylated monoclonal antibodies directed against cell surface antigens. This combination permitted flow cytometric control of the magnetic separation. Ficoll-Paque-separated peripheral blood mononuclear cells were first eliminated from monocytes by leucine-methyl ester treatment. B cells were enriched to 97% after magnetic depletion of CD3-positive T cells and magnetic enrichment of CD20-positive B cells. The separated B cells could be induced to proliferation and antibody production by various in vitro stimuli.
Abts
H
H
I. Med. Klinik, Universität Köln, Cologne, F.R.G.
Emmerich
M
M
Miltenyi
S
S
Radbruch
A
A
Tesch
H
H
D000921:Antibody-Producing Cells / Q000166:cytology* / Q000276:immunology; D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis*; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002454:Cell Differentiation; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D008280:Magnetics; D009000:Monocytes / Q000166:cytology; D013601:T-Lymphocytes / Q000166:cytology
D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte