Separation of Viable Leukocytes from Normal Human Blood

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Abstract
A method for the separation of viable leukocytes from normal human blood is described. It is based on sedimentation of the erythro-cytes with polyvinylpyrrolidone and subsequent lysis of the remaining erythrocytes with streptolysin-O. A mean of 57% of the leukocytes initially present in the samples could be recovered. The ratio of erythrocytes to leukocytes was far below 1:1, more likely 1:50 or less after completion of the separation. The rather good percentage of recovery was aided to some extent by diluting the initial blood sample 1:10 before the sedimentation. Viability studies after the separation was completed included measurements of the leukocytes uptake of oxygen, dyeexclusion tests, observation of division in short-term cultures, as well as attachment of the cells and ameboid movements. All parameters thus observed indicated that the separation procedure induced no gross alterations in the cells, and certainly no serious loss of viability.