Abstract

This paper describes a technique for the isolation of lymphocytes from haparinized human peripheral blood. When whole blood was layered on top of a mixture of Ficoll and Isopaque and gradient centrifuged at room temperature, the cellular elements of whole blood were divided into two main parts, a lymphocyte bone in plasma layer and a layer consisting of sedimented granulocytes and erythrocytes at the bottom of the tube. By this method the isolation of viable lymphocyte with 96.6% purity was possible in a short period of time. Impurities found in the lymphocyte zone was granulocytes; 0.2%, monocytes; 2.8%, eosinophils; 0.4%, erythrocytes; less than 5% of total cell counts respectively. Isolating conditions and purities together with detailed directions are presented. It is shown in the present paper that adopting the procedure introduced a lymphocyte suspension of extreme purity with high viability may be obtained for lymphocyte research.