A simple and rapid method for isolation of eosinophilic granulocytes from human blood

• Published in: Journal of leukocyte biology Vol. 52; no. 3; pp. 331 - 336
• Main Authors: Cramer, Rita, Dri, Pietro, Zabucchi, Giuliano, Patriarca, Pierluigi
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Society for Leukocyte Biology 01.09.1992; Federation of American Societies for Experimental Biology
• Subjects: Biological and medical sciences; cell density; cell separation; Cell Separation - methods; Centrifugation, Density Gradient - methods; eosinophilia; eosinophils; Eosinophils - cytology; Eosinophils - physiology; Erythrocytes - cytology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; Molecular immunology; Neutrophils - cytology; Povidone; Respiratory Burst; Silicon Dioxide; superoxide anion; Techniques; Human; Rapid technique; Purification; Granulocyte; Isolation; Neutrophil; Eosinophil; Comparative study; Cell density
• ISSN: 0741-5400; 1938-3673
• DOI: 10.1002/jlb.52.3.331

A simple and rapid method for the purification of morphologically and functionally intact eosinophils from human blood of both normal and eosinophilic subjects is described. The method is based on a single centrifugation of total blood leukocytes suspended in Percoll with specific gravity 1.0853 g/ml, after erythrocyte removal by dextran sedimentation. The peculiarity of this isolation technique is the maintenance of strictly physiological values of pH and osmolality throughout the entire procedure. Moreover, the cells are not subjected to measures aimed at changing the physical properties of either neutrophils or eosinophils. Because of such characteristics, this isolation method could be usefully exploited for comparative studies of normal and eosinophilic nor‐modense eosinophils and of neutrophils and eosinophils from the same noneosinophilic subject.