Abstract
Abstract Methods are presented for obtaining highly purified preparations of basophilic leukocytes from the blood (mean purity, 62%) and bone marrow (mean purity, 38%) of guinea pigs. The following steps are performed in sequence: harvest of blood or bone marrow cells from animals primed with antigen to develop a basophilia; phagocytosis of carbonyl iron particles by neutrophils and monocytes; sedimentation of erythrocytes with gelatin (in the case of peripheral blood); removal of phagocytic cells and residual-free iron particles by passage through a magnetic field; centrifugation on a newly developed 4.5 to 9.5% linear Ficoll density gradient; and removal of residual erythrocytes by Tris-NH4Cl (optional step for peripheral blood). The basophils isolated are viable, functional cells, and, depending on the source and degree of purity desired, may be obtained in quantities of 5 × 106 to 2 × 108 and in an overall yield of 25 to 100%.
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