Human stored platelet function in different extracellular environments. Human platelets were collected by apheresis and stored as RTP and CSPs for 7 days. Testing was performed after 3 days and at the end of storage (7 days). (A) Outline of the sequence of study events. (B-C) Light transmission aggregometry data (shown as % of maximum aggregation) using either RTP (red dots) or CSP (blue dots) diluted either with FFP or using concomitant stored plasma (ExE). Platelets were stimulated with either 20μM ADP (B) or 5 μg/mL of collagen (C). (D-K) The same groups as outlined above were tested using flow cytometry for αIIbβ3 integrin activation by adding PAC1 antibody (D,F,H,J) or for α-degranulation by adding P-selectin antibody (E,G,I,K) (all data acquired as median MFI). Platelets were tested as baseline (D-E) (ie, no agonist) or after the addition of 20μM ADP (F-G), 1mM Par4-P (H-I), and 100 ng/mL of CVX (J-K). Graphs show individual data points (n = 4-5). ∗P ≤ .05; ∗∗P ≤ .01; ∗∗∗P ≤ .001. Lines between dots have been added to identify individual data points between fresh and dilution with ExE and FFP and do not represent continuity or timeline. Statistical analysis with 1-way analysis of variance (ANOVA), with Tukey correction for multiple comparisons. D3, day 3; D7, day 7; MFI, mean fluorescence intensity; PLT, platelet.

Human stored platelet function in different extracellular environments. Human platelets were collected by apheresis and stored as RTP and CSPs for 7 days. Testing was performed after 3 days and at the end of storage (7 days). (A) Outline of the sequence of study events. (B-C) Light transmission aggregometry data (shown as % of maximum aggregation) using either RTP (red dots) or CSP (blue dots) diluted either with FFP or using concomitant stored plasma (ExE). Platelets were stimulated with either 20μM ADP (B) or 5 μg/mL of collagen (C). (D-K) The same groups as outlined above were tested using flow cytometry for αIIbβ3 integrin activation by adding PAC1 antibody (D,F,H,J) or for α-degranulation by adding P-selectin antibody (E,G,I,K) (all data acquired as median MFI). Platelets were tested as baseline (D-E) (ie, no agonist) or after the addition of 20μM ADP (F-G), 1mM Par4-P (H-I), and 100 ng/mL of CVX (J-K). Graphs show individual data points (n = 4-5). ∗P ≤ .05; ∗∗P ≤ .01; ∗∗∗P ≤ .001. Lines between dots have been added to identify individual data points between fresh and dilution with ExE and FFP and do not represent continuity or timeline. Statistical analysis with 1-way analysis of variance (ANOVA), with Tukey correction for multiple comparisons. D3, day 3; D7, day 7; MFI, mean fluorescence intensity; PLT, platelet.

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