Separation of human B lymphocytes by combined rosetting with mouse and sheep red cells.
A technique for obtaining highly enriched B lymphocyte subpopulation from isolated peripheral blood lymphocytes is described. The method consists in combined use of mouse and sheep red cell rosettes. In the first step of the method, B cells-containing fraction of lymphocytes and papainized mouse red cells form rosettes which are separated by Ficoll-Hypaque gradient centrifugation. In the second step, after lysis of mouse erythrocytes in the sediment, the recovered B cell rich fraction is purified by E rosetting procedure, B lymphocyte subpopulation obtained in this way, presents a good viability and is almost completely pure.
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D003601:Cytotoxicity Tests, Immunologic; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology; D006801:Humans; D051379:Mice; D010206:Papain / Q000494:pharmacology; D012397:Rosette Formation; D012756:Sheep
D010206:Papain