The Isopaque-Ficoll method re-evaluated: selective loss of autologous rosette-forming lymphocytes during isolation of mononuclear cells from human peripheral blood.
A subpopulation of lymphocytes is defined, which--under conditions used for isolating mononuclear cells from peripheral blood by the Isopaque-Ficoll gradient--is lost to the erythrocyte pellet because of rosette formation with the autologous erythrocytes in the gradient. Reapplication of the resuspended erythrocyte pellet on a new gradient with higher density could, however, recover the subpopulation. Lymphocyte surface marker analysis on such cells showed that they were enriched in T cells (judged by rosette formation with sheep erythrocytes) and in autorosette-forming cells. The importance of these findings is discussed in relation to the wide use of the Isopaque-Ficoll method.
Hokland
P
P
University Department of Medicine and Haematology, Aarhus, Denmark.
Heron
I
I
D002469:Cell Separation / Q000379:methods; D002498:Centrifugation; D005362:Ficoll; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D008794:Metrizoic Acid; D012397:Rosette Formation
D005362:Ficoll; D008794:Metrizoic Acid