Leucocyte energy metabolism. VI. Simultaneous and improved isolation of lymphocytes and polymorphonuclear leucocytes from a single of human blood.
A technique, which combines a Ficoll-Rompacon treatment, sedimentation in a Dextran medium and erythrolysis by NH4C1, permitting the simultaneous isolation of polymorphonuclear cells (PMN) and lymphocytes from a single sample of human blood is described. The contamination of the PMN suspensions by other blood cells, including other categories of white cells, is minimal. However, the lymphocyte preparations, free of other white cells, are still contaminated by a non-negligible amount of thrombocytes. It could be shown that the biochemical system studied, the functional behavior and the morphological state of these cells are similar to that of white cells purified with the procedures previously used. Further, a numeration technique, based on DNA estimation, is presented which can be used when the cell count becomes inaccurate by cell agglutination.
D002469:Cell Separation / Q000379:methods; D006019:Glycolysis; D006593:Hexokinase / Q000097:blood; D006801:Humans; D007773:Lactates / Q000097:blood; D007962:Leukocytes / Q000166:cytology / Q000201:enzymology*; D008214:Lymphocytes / Q000166:cytology / Q000201:enzymology*; D009504:Neutrophils / Q000166:cytology / Q000201:enzymology*; D010732:Phosphofructokinase-1 / Q000097:blood; D010735:Phosphoglycerate Kinase / Q000097:blood; D011770:Pyruvate Kinase / Q000097:blood
D007773:Lactates; D006593:Hexokinase; D010732:Phosphofructokinase-1; D011770:Pyruvate Kinase; D010735:Phosphoglycerate Kinase