Large Scale Preparation of Leukocyte Concentrates and Further Purification of Lymphocytes and Granulocytes with the IBM 2991 Cell Processor

Abstract

A procedure is described for the large-scale production of pure suspensions of human lymphocytes and granulocytes. Platelet-poor buffy-coats, obtained by centrifugation of six units of ABO-compatible ACD blood, were pooled and centrifuged again, resulting in a crude leukocyte concentrate containing approximately 60 per cent of the original amount of leukocytes. Further fractionation of such crude leukocyte concentrates was achieved by scaling up Bøyum's Ficoll-Isopaque density gradient technique with the aid of an IBM 2991 blood cell processor. In this way 2.9 +/- 0.3 X 10(9) mononuclear leukocytes, consisting of 90 +/- 0.7 per cent lymphocytes and 5.3 +/- 0.3 X 10(9) polymorphonuclear leukocytes of 92 +/- 0.7 per cent purity could be isolated in one run. The overall yield was 36 per cent of the original amount of leukocytes.