The effect of centrifugation speed and time on pre-analytical platelet activation

Söderström, Anna; Nybo, Mads; Nielsen, Christian; Vinholt, Pernille Just

doi:10.1515/cclm-2016-0079

Cited by 55 publications

(37 citation statements)

References 32 publications

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“…The platelet aggregates detected in the flow‐cytometric assay might be too small to change absorbance readings in LTA, and would therefore remain undetected by LTA. The results from this platelet aggregation assay might not be representative of platelet function in whole blood, as large platelets are removed during centrifugation . Also, the results are based on a fraction of platelets in the mix.…”

Section: Discussionmentioning

confidence: 99%

Measurement of platelet aggregation, independently of patient platelet count: a flow‐cytometric approach

Vinholt

Frederiksen

Hvas

et al. 2017

Journal of Thrombosis and Haemostasis

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Background Methods for testing platelet aggregation in thrombocytopenia are lacking. Objective To establish a flow-cytometric test of in vitro platelet aggregation independently of the patient's platelet count, and examine the association of aggregation with a bleeding history in thrombocytopenic patients. Patients/methods We established a flow-cytometric assay of platelet aggregation, and measured samples from healthy individuals preincubated with antiplatelet drugs, and samples from two patients with inherited platelet disorders. Then, we included 19 healthy individuals and 20 patients with platelet counts of ≤ 50 × 10 L , diagnosed with acute myeloid leukemia or myelodysplastic syndrome. We measured platelet aggregation and platelet activation by platelet surface expression of activated glycoprotein IIb-IIIa, P-selectin and CD63 after addition of agonists: collagen-related peptide, thrombin receptor-activating peptide (TRAP), and ADP. Results The platelet aggregation assay showed a low intraserial coefficient of variation of ≤ 3%. Similar results were obtained for platelet-rich plasma and isolated platelets at platelet counts of > 10 × 10 L ; otherwise, platelet isolation was required. The platelet aggregation percentage decreased with increasing antiplatelet drug concentration. Platelet aggregation in patients was reduced as compared with healthy individuals: 42% (interquartile range [IQR] 27-58) versus 66% (IQR 60-67) for TRAP; 41% (IQR 25-48) versus 70% (IQR 69-72) for collagen-related peptide; and 44% (IQR 30-53) versus 65% (IQR 46-72) for ADP. Platelet activation after stimulation was reduced in patients and correlated with platelet aggregation (e.g. r = 0.78-0.81 when stimulated with collagen-related peptide). Platelet aggregation had a negative predictive value of 100% for a bleeding tendency among patients. Conclusion The established platelet aggregation assay was applicable for thrombocytopenic patients, and improved the identification of bleeding risk.

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Section: Discussionmentioning

confidence: 99%

Measurement of platelet aggregation, independently of patient platelet count: a flow‐cytometric approach

Vinholt

Frederiksen

Hvas

et al. 2017

Journal of Thrombosis and Haemostasis

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show abstract

“…38 In our assay, special care was undertaken to minimize platelet activation prior to coating the plates which included keeping centrifugation at a setting shown to minimize premature activation. 39 However, if some degree of activation occurred, results were reproducible in independent experiments using different SDPs to coat plates which may indicate that binding occurred onto platelets and not just to mediators released as a result of activation. Nevertheless, this is an area that will require future investigation.…”

Section: Discussionmentioning

confidence: 95%

Use of a whole‐cell ELISA to detect additional antibodies in setting of suspected heparin‐induced thrombocytopenia

Bashover

Stefaniuk

Maitta

2019

European J of Haematology

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Objectives: Type II heparin-induced thrombocytopenia (HIT) is mediated by formation of antibodies to platelet factor 4 (PF4)-heparin complexes. We evaluated anti-PF4heparin-negative samples for the presence of additional anti-platelet and anti-red blood cell (RBC) antibodies using whole-cell platelet/ RBC ELISAs we developed. Methods: Seventy-three samples tested for anti-PF4-heparin by ELISA were included: 62 tested negative, 9 tested positive, and 2 had equivocal results. Plasma specimens from healthy donors were used as controls.Results: 100% (9/9) anti-PF4-positive samples had anti-platelet antibodies detected by whole-cell platelet ELISA. 42.2% (27/64) anti-PF4-heparin-negative samples were negative for anti-platelet and anti-RBC antibodies. 32.8% (21/64) negative samples showed reactivity to both platelets and RBC; 12.5% (8/64) negative samples were each reactive with either platelet or RBC ELISA, respectively. Additionally, two samples that tested equivocal by anti-PF4-heparin ELISA had antibodies to both platelets and RBC by whole-cell ELISA. Conclusions:Our study suggests that patients with thrombocytopenia testing negative for anti-PF4-heparin may still harbor antibodies to platelets. However, additional research is needed to determine the significance of these antibodies. Nevertheless, these findings may encourage clinicians to further investigate patients with possible immune-mediated etiologies of thrombocytopenia and anemia. K E Y W O R D Santi, antibodies, heparin-induced thrombocytopenia, PF4-heparin, platelet, red cells, wholecell ELISA

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“…28,29 Apart from these there was no well-proven study in which activation of PRP was mentioned and whether it was activated or not. There were some preclinical 30 and clinical 31,32 studies, but there was no consensus on a protocol. In literature, it was reported that early immature activation and growth factors release during the preparation of PRP pose other problems.…”

Section: Discussionmentioning

confidence: 99%

Is there a treatment protocol in which platelet-rich plasma is effective?

Yılmaz

Akkaya

Isyar³

et al. 2016

Journal of Orthopaedics

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The effect of centrifugation speed and time on pre-analytical platelet activation

Abstract: Proportional to centrifugation speed, platelets in plasma and platelet-rich plasma were activated with centrifugation speed, cell content and composition changed while platelet aggregation was unaltered.

Cited by 55 publications

References 32 publications

Measurement of platelet aggregation, independently of patient platelet count: a flow‐cytometric approach

Measurement of platelet aggregation, independently of patient platelet count: a flow‐cytometric approach

Use of a whole‐cell ELISA to detect additional antibodies in setting of suspected heparin‐induced thrombocytopenia

Is there a treatment protocol in which platelet-rich plasma is effective?

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