TABLE 6

Effects Produced by Anticoagulants or Processing Techniques on Platelet Isolation

Cell TypeAdverse Effects
Platelets
 Effect on cell morphology• EDTA: induces platelet swelling, changes in size distribution, and accelerates shape changes137
 Effect on cell chemistry• Heparin: increases the number of platelet-monocyte aggregates compared with PPACK, sodium citrate, and EDTA32
 Effect on sample handling or collection• EDTA and citrate: differentiate more accurately the number of single cells that are present in the sample from clusters137 by dispersing and decreasing the number of reversible platelet-neutrophil aggregates,32 or the number of platelet-monocyte aggregates compared with PPACK,32 or compared with hirudin and heparin.138
 Effect on cell chemistry and on sample handling or collection• EDTA and EGTA: also reduce the number of platelet-leukocyte aggregates32
• EDTA: decreases viability of platelets compared with acid citrate dextrose137
• PPACK: the preferred anticoagulant for platelet isolation due to less platelet activation32
• Citrate: the preferred anticoagulant in the event of a predicted delay to immunostaining and fixation,32 but can produces cell aggregates137
• Platelet-monocyte aggregates remain longer in peripheral blood, and they can be quantified by flow cytometric analysis32
• BAPA (dual inhibitor of factor Xa and thrombin): recommended when delays are expected between blood collection by venipuncture and transportation to remote locations; maintains stable platelet function response (IPA and ATP secretion) and cell dense granule secretion up to 48 h at RTa compared with sodium citrate139
• Platelet-monocyte aggregates increase in sample from intravenous cannulae compared with sample obtained by venipuncture32
• Citrate-phosphate-dextrose: concentrations of lysolecithins (lysoPCs, compounds derived from phosphatidylcholines) increase in these platelets stored for 5–7 d compared with the fresh platelet sample106
  • ATP, adenosine-5′-triphosphate; BAPA, benzylsulfonyl-D-Arg-pro-4-amidinobenzylamide; EGTA, ethylene glycol tetraacetic acid; IPA, Impedance platelet aggregation; PPACK, D-phenylalanyl-l-prolyl- l -arginine chloromethyl ketone.

  • a RT = 20–25°C.