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.