OPTIMIZED LYMPHOCYTE ISOLATION - ONE-STEP PROCEDURE FOR ISOLATION OF HUMAN-LYMPHOCYTES BY MEANS OF NYCOPREP MIKXER - RECOVERY AND PURITY INCOMPARISON WITH OTHER SEPARATION MEDIA

The separation of human lymphocytes by mixing of the blood sample with a Nycodenz medium (NycoPrep Mixer) prior to centrifugation avoids the time consuming overlayering of gradients. The cell yields of four sep aration media (housemade Ficoll-Visotrast-Mixture, Lymphoprep, NycoPre p 1.077, NycoPrep Mixer) evaluated were not significant different. The cell suspensions obtained by the one-step method were hardly contamin ated with platelets and erythrocytes, showed the lowest content of mon ocytes but the comparably highest content of lymphocytes. No differenc es were seen in the distribution of T-cells and T cell subpopulations (CD4, CD8) measured by flow cytometry, only the B cell part was slight ly reduced using Nycodenz media. HLA-typing showed identical pattern i n all suspensions tested. This method can be recommended particularly for the handling of many as well as large blood samples.