TABLE 5

Effects Produced by Anticoagulants or Processing Techniques on White Blood Cell Isolation

Cell TypeAdverse Effects
Leukocytes
 Effect on cell morphology• EDTA: excess EDTA causes leukocytes membrane damage126
 Effect on cell chemistry• EGTA and oxalate: produce lower yield of cells130
• Heparin: at concentrations >20 IU/mL decreases migration and chemotaxis130
PMNs
 Effect on cell morphology and on sample handling or collection• EDTA: vacuolization of PMNs occurred after 3–4 h of storage at RTa and increased after 6 h131; no or minimal changes were found when samples were storage at 4°C132
 Effect on cell chemistry• EDTA: results in an increase in the number of PMNs isolated and a decrease in PMN activation, as compared with sodium citrate or heparin;133 PMN count remains stable for ≥3 d with EDTA when whole blood is stored at RT134
• Heparin and sodium citrate: decreases the efficiency of PMN recovery133
• Heparin: increases PMN activation due to stimulation of adhesion molecules such as integrins135
Monocytes
 Effect on cell morphology and on sample handling or collection• EDTA: vacuolization of monocytes occurred after 1 h of storage at RT and increased after 4 h131; no or minimal changes were found when samples were stored at 4°C132
 Effect on cell chemistry• EDTA and heparin: increases the number of monocytes isolated, but they did not affect monocyte viability, spontaneous migration, chemotaxis, phagocytosis, or in killing Candida albicans130
Lymphocytes
 Effect on cell chemistry• Sodium citrate: lymphocytes appear stable25
• Heparin: inhibits proliferation of lymphocytes27
PBMC (lymphocytes and monocytes)
 Effect on sample handling or collection• BD Vacutainer CPT cell preparation tube with sodium citrate or heparin: allows for separation of plasma and PBMCs from erythrocytes and granulocytes; very efficient for collection and cryopreservation of active PBMCs136
  • PBMC, peripheral blood mononuclear cell; PMN, polymorphonuclear neutrophil.

  • a RT = 20–25°C.