INVESTIGATION ON THE METHOD OF LEUKOCYTE SEPARATION

Abstract

The establishment of the excellent leukocyte separation method is at first required in biochemical studies of leukocyte enzymes. The leukocyte separation method with highmolecular dextran (average M.W. 228, 000) was examined following the studies of phytohemagglutinin and Gam Acasia methods formerly reported. Several kinds of concentrations of dextran solution, mixing ratios of dextran solution and blood, and influence of temperature were investigated in order to establish optimal conditions i.e. comparative leukocyte yields, convenience, and minimal erythrocyte contaminations. It was found that the condition of mixing blood and six per cent dextran solution in the following volume ratio (B: D) 6: 1 was most suitable. The mean of leukocyte yields was 68.3±0.55 and erythrocyte contaminations were below 94 per cent of leukocytes. The nature of separated leukocytes showed no significant changes comparing with that of fresh capillar blood, by Giemsa staining, supravital staining, phagocytosis-test of indian ink and differentials. These indicate that leukocyte injuries with dextran method are minimal and separated leukocytes are usable for biochemical studies. The dextran method is better than the phytohemagglutinin and Gam Acasia methods in regard to leukocyte yields and convenience.