Impact of buffy coat storage on the generation of inflammatory cytokines and platelet activation.
BACKGROUND
Whole blood-derived buffy coat (BC) has become an alternative source from which to prepare random-donor platelet concentrates. The influence of prolonged storage of BC prior to platelet concentrate preparation is a matter of controversy. The impact of BC storage on cytokine release was evaluated and the platelet activation quantified.


METHODS
BCs were prepared from whole-blood donations after hard-spin centrifugation. After 1, 3, 6, 12, and 24 hours of storage at 22 degrees C without agitation, samples were withdrawn for cell count and blood gas analysis and measurement of interleukin (IL)-1beta, IL-6, IL-8, tumor necrosis factor alpha and platelet factor 4. Platelet surface markers CD41a, CD42b, CD62P, and CD63 were analyzed by flow cytometry, and the antibody-binding sites were quantified by using microbeads.


RESULTS
Inflammatory cytokines IL-1beta, IL-6, and tumor necrosis factor alpha were hardly detectable in stored BCs but levels of IL-8 increased in 25 percent of BCs after 24 hours. A constant increase in platelet factor 4 was observed, which accelerated after 12 hours of storage. Analysis of platelet surface markers showed an initial decrease of platelet activation, followed by an increase after storage for 12 to 24 hours.


CONCLUSIONS
Storage of BCs for up to 12 hours without agitation showed a good preservation of platelets but storage of BCs for 24 hours resulted in increased platelet activation and significantly higher release of platelet factor 4 and IL-8. Stored BCs might well be suitable for platelet preparation, but their storage time should not greatly exceed 12 hours.
Klüter
H
H
Institute of Immunology and Transfusion Medicine, University of LübeckSchool of Medicine, Germany.
Schlenke
P
P
Müller-Steinhardt
M
M
Paulsen
M
M
Kirchner
H
H
D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology; D001666:Binding Sites, Antibody / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D001784:Blood Gas Analysis; D001792:Blood Platelets / Q000276:immunology; D001793:Blood Preservation; D016207:Cytokines / Q000096:biosynthesis*; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007958:Leukocyte Count; D007962:Leukocytes; D019007:P-Selectin / Q000276:immunology; D015539:Platelet Activation / Q000502:physiology*; D010976:Platelet Count; D010978:Platelet Factor 4 / Q000378:metabolism; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000276:immunology; D010980:Platelet Membrane Glycoproteins / Q000276:immunology; D060149:Tetraspanin 30; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015415:Biomarkers; C554586:CD63 protein, human; D016207:Cytokines; D007375:Interleukin-1; D015850:Interleukin-6; D019007:P-Selectin; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D014409:Tumor Necrosis Factor-alpha; D010978:Platelet Factor 4