Cell Adhesion to Immobilized Platelets Parallel-Plate Flow Chamber

Cell Adhesion to Immobilized Platelets Parallel-Plate Flow Chamber 

In this assay, immobilized platelets are pretreated with a GPIIb/IIIa antagonist, and any unbound drug is washed away before the perfusion of monocytic THP-1 cells. McCarty et al. (2004) demonstrated that agents with slow platelet off-rates such as XV454 (ti/2 of dissociation = 110 min Kd = 1 nM) and abciximab (ti/2 of dissociation = 40 min Kd = 9.0 nM) that are distributed predominantly as receptor-bound entities with little unbound in the plasma, can effectively block these heterotypic interactions as shown by Abulencia et al. (2001) and by Mousa et al. (2002). In contrast, agents with relatively fast platelet dissociation rates such as orbofiban (ti/2 of dissociation = 0.2 min Kd 110 nM), whose antiplatelet efficacy depend on the plasma concentration of the active drug, do not exhibit any inhibitory effects as described by Mousa etal. (2002). 

Soak glass coverslips overnight in 70 % nitric acid. 

Dry coverslips by washing once with acetone, followed by immersion in a 4 % solution of APES in acetone for 2 min. 

Repeat Step 3, followed by a final rinse of the glass coverslips with acetone. 

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