Chemotaxis as a preparative technique for human polymorphonuclear leukocytes

Abstract

Human peripheral blood phagocytic cells were prepared in large quantities utilizing their chemotactic characteristics. Cells were isolated using an enlarged version of the Boyden chemotactic chamber and were collected after migrating through a Millipore membrane into a solution containing chemotactic factor. Chemotactic factors consisted of normal human serum, casein-saturated medium, and a combination of human serum and casein-saturated medium. The most effective attractant was the mixture of casein and fresh human serum. Yields of up to 24.8 times 10(6) cells could be attained from a starting volume of approximately 12 c.c. of peripheral blood. These cells were greater than 95 per cent polymorphonuclear leukocytes with the remaining cells being monocytes. No erythrocytes or lymphocytes contaminated these preparations. The advantages of this technique are as follows: (1) large quantities of human peripheral blood phagocytes consisting of greater than 95 per cent polymorphonuclear cells can be obtained: (2) these cells are known to be biologically active sice chemotaxis is a requirement for their isolation; (3) the separative method is dependent on chemotactic properties rather than sedimentation characteristics; and finally (4) the resulting cell preparations are virtually devoid of lymphocytes and erythrocytes.