Evaluation of the Total Thrombus-Formation System (T-TAS): application to human and mouse blood analysis: evaluation of T-TAS in assessing clot formation

Rashid Hafidh Rashid Al Ghaithi, Jun Mori, Zoltan Nagy, Annabel Maclachlan, Lewis Hardy, Helena Philippou, Emma Hethershaw, Neil Morgan, Yotis Senis, Paul Harrison

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The Total Thrombus-formation Analyser System (T-TAS) is a whole blood flow chamber system for the measurement of in vitro thrombus formation under variable shear stress conditions. Our current study sought to evaluate the potential utility of the T-TAS for the measurement of thrombus formation within human and mouse whole blood. T-TAS microchips (collagen, PL chip; collagen/tissue thromboplastin, AR chip) were used to analyse platelet or fibrin-rich thrombus formation respectively. Blood samples from humans (healthy and patients with mild bleeding disorders) and wild-type (WT), mice were tested. Light transmission lumi-aggregometer (lumi-LTA) was performed in PRP using several concentrations of ADP, adrenaline, arachidonic acid, collagen, PAR-1 peptide and ristocetin. Thrombus growth (N=22) increased with shear within PL (4:40±1.11, 3:25±0.43 and 3:12±0.48 mins [1000, 1500 and 2000s-1]) and AR chips (3:55±0.42 and 1:49±0.19 [240s-1 and 600s-1]). The area under the curve (AUC) on the PL chip was also reduced at 1000s-1 compared to 1500/2000s-1 (260±51.7, 317± 55.4 and 301±66.2 respectively). In contrast, no differences in the AUC between 240s-1 and 600s-1 were observed in the AR chip (1593±122 and 1591±158). The intra-assay coefficient of variation (CV) (n=10) in the PL chip (1000s-1) and AR chip (240s-1) were T1014.1%, T6016.7%, T10-6022.8% and AUC1024.4% or T10 9.03%, T808.64%, T10-8023.8% and AUC305.1%. AR chip thrombus formation was inhibited by rivaroxaban (1µM), but not with ticagrelor (10µM). In contrast, PL chip thrombus formation was totally inhibited by ticagrelor. T-TAS shows an overall agreement with lumi-LTA in 87% of patients (n=30) with normal platelet counts recruited into the genotyping and phenotyping of platelet (GAPP) study and suspected to have a platelet function defect. The onset (T10) of thrombus formation in WT mice (N= 4) was shorter when compared to humans e.g. PL chip (1000s-1) T10 were 02:02±00:23 and 03:30±0:45 respectively). T-TAS measures in vitro thrombus formation and can be used for monitoring antithrombotic therapy, investigating patients with suspected platelet function defects and monitoring platelet function within mice.
Original languageEnglish
Number of pages22
Early online date26 Oct 2018
Publication statusE-pub ahead of print - 26 Oct 2018


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