Preparation and culture of human lymphocytes

Abstract

This unit presents protocols for preparation of lymphocytes from peripheral (whole) blood or leucopherisis samples, first by differential centrifugation in Ficoll-Hypaque to separate them from erythrocytes and granulocytes. Repeated centrifugation removes the plasma and platelets. Monocyte/macrophage cells and dendritic cells can be purified from the T and B cell population by exploiting their adhesion to serum-coated plates in the presence of recombinant IL-3. Different subpopulations of lymphocytes can be isolated by specific antibodies bound to magnetic beads.