platelet adhesion & aggregation

Overview

Platelet adhesion, aggregation and thrombi formation (blood clot) can easily be studied using Cellix's microfluidic pumps and biochips.  The dimensions of the biochips can mimic those of human capillaries thereby enabling researchers to simulate in vivo microenvironments. A key additional benefit of using microfluidics is the reduction in sample volume required for experiments.  This is particularly important when using whole blood.  Typically a sample of 10-40mL is taken from donors in vacutainers which are citronated to prevent clotting.  Despite the treatment, a researcher only has limited time (2-3 hours) before the blood will inevitably clot.  Therefore speed and throughput are vital.  Cellix's 8-channel Mirus Evo Nanopump enables researchers to conduct up to 8 assays simultaneously in microfluidic biochips which may be pre-coated with VWF, collagen, fibrinogen or other proteins of interest.  Cellix's Vena8 Fluoro+ biochip is compatible with confocal microscopy making it particularly attractive for applications in thrombosis.

Assays:  

  • platelet observation under shear flow
  • platelet rolling, adhesion & aggregation assays
  • platelet-endothelial cell interaction studies
  • investigation of anticoagulant / anti-platelet therapies

 

Assay Examples:

 

Platelet adhesion profile to VWF and Collagen:

Coat Vena8 Fluoro+ biochip with VWF or collagen according to standard protocol.
Connect to Mirus Evo Nanopump and use the standard protocol for thrombosis assays.
Results:  Samples treated with EP4 agonist ONO AE1-329 or PGE2 largely prevented thrombus formation; only small aggregates were observed.  Effects of EP4 agonist and PGE2 were reversed by the EP4 antagonist ONO AE3-208.

 


 

Platelet adhesion profile to fibrinogen vs. whole blood sample treated with ReoPro:

Coat Vena8 Fluoro+ biochip with fibrinogen according to standard protocol.
Connect to Mirus Evo Nanopump and use the standard protocol for thrombosis assays.
Results: ReoPro (abciximab), a glycoprotein IIb/IIIa receptor antagonist, clearly inhibits platelet adhesion to fibrinogen.

 


 

The Prostaglandin E2 Receptor EP4 is Expressed by Human Platelets and Potently Inhibits Platelet Aggregation and Thrombus Formation

Coat Vena8 Fluoro+ biochip with protein according to standard protocol.
Connect to Mirus Evo Nanopump and use the standard protocol for thrombosis assays.
Results: ReoPro (abciximab), a glycoprotein IIb/IIIa receptor antagonist, clearly inhibits platelet adhesion to fibrinogen.

 

 


 

A small-molecule inhibitor of integrin α2β1 introduces a new strategy for antithrombotic therapy.

Coat Vena8 Fluoro+ biochip with collagen according to standard protocol.
Connect to Mirus Evo Nanopump and use the standard protocol for thrombosis assays.
Results:

  • • BTT-3016 inhibited the wt adhesion of platelets on collagen by 65% but did not affect the deposition of α2-/- mouse-derived platelets.
  • • Platelets from α2-/- mice showed weaker attachment on collagen-coated capillaries in comparison with the platelets from wt mice.