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Purification of Basophilic Leukocytes from Guinea Pig Blood and Bone Marrow¹

From the Departments of Pathology, Massachusetts General Hospital and Harvard Medical School and Tufts University Medical School, Boston, Massachusetts 02114

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 x 10⁶ to 2 x 10⁸ and in an overall yield of 25 to 100%.

Footnotes

¹ This work was supported by United States Public Health Service Grants AI-09529 and CA-15136 and by American Cancer Society Grant IM-44.

² H. F. D. is United States Public Health Service Research Career Development Awardee 1-K-04-AI-46352.