title: A rapid method for isolation of human mononuclear cells free of significant platelet contamination.
abstract: In order to obtain mononuclear cells from peripheral human blood for the study of cell surface receptors, it was necessary to effectively eliminate contaminating platelets. The usual Hypaque-Ficoll isolation procedures were found to produce mononuclear cells contaminated with 10-1000 platelets per mononuclear cell (by phase microscopy). Multiple slow speed centrifugations reduced the contamination to 5-10 platelets per mononuclear cell. However, centrifugation of EDTA-anticoagulated blood through Hypaque (D20(20) 1.060) at 400 X g for 5 min at 22 degrees C followed by the usual Hypaque-Ficoll gradient reduced platelet contamination to less than 1 platelet per 2 mononuclear cells. Thus, a rapid and simple gradient procedure is capable of significantly reducing platelet contamination of mononuclear cell preparations and should facilitate the analysis of mononuclear cell receptors and functions.
mesh_terms: D001792:Blood Platelets; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods; D003973:Diatrizoate / Q000494:pharmacology; D003974:Diatrizoate Meglumine / Q000494:pharmacology; D004338:Drug Combinations / Q000494:pharmacology; D005362:Ficoll / Q000494:pharmacology; D006801:Humans; D008214:Lymphocytes; D009000:Monocytes; D011941:Receptors, Adrenergic / Q000032:analysis
chemical_list: D004338:Drug Combinations; D011941:Receptors, Adrenergic; D003973:Diatrizoate; D005362:Ficoll; D003974:Diatrizoate Meglumine; C027278:urovision