Using Well-Plate Microfluidic Devices to Conduct Shear-Based Thrombosis Assays

Shear stress plays a critical role in regulating platelet adhesion and thrombus formation at the site of vascular injury. As such, platelets are often examined in vitro under controlled shear flow conditions for their hemostatic and thrombotic functions. Common shear-based platelet analyses include the evaluation of genetic mutants, inhibitory or experimental compounds, matrix substrates, and the effects of different physiological and pathological shear forces. There are several laboratory instruments widely used for studying shear flow, including cone and plate viscometers and parallel plate perfusion chambers. These technologies vary widely in the types of samples, substrates, blood volumes, and throughput that are involved. Here, we describe a microfluidic system for platelet analysis under shear flow. We used the devices to study thrombus formation on collagen I and von Willebrand factor. The system was also used to investigate dose response to the antiplatelet compound, Abciximab, under shear flow conditions with an emphasis on maximizing the number of data points per single patient sample. The presented method confers multiple advantages over conventional approaches. These include the ability to assess up to 24 conditions simultaneously in real time, maintain identical physical conditions across experiments, and use extremely low donor volumes.

[1]  Carolyn G. Conant,et al.  Wound Healing Assays in Well Plate—Coupled Microfluidic Devices with Controlled Parallel Flow , 2010 .

[2]  Takashi Nakamura,et al.  Platelet Adhesion to Native Type I Collagen Fibrils , 1998, The Journal of Biological Chemistry.

[3]  L. McIntire,et al.  Ristocetin-dependent, but not botrocetin-dependent, binding of von Willebrand factor to the platelet glycoprotein Ib-IX-V complex correlates with shear-dependent interactions. , 2001, Blood.

[4]  J. Sixma,et al.  A new perfusion chamber to detect platelet adhesion using a small volume of blood. , 1998, Thrombosis research.

[5]  A. Groisman,et al.  Microfluidic devices for studies of shear-dependent platelet adhesion. , 2008, Lab on a chip.

[6]  Scott L Diamond,et al.  A membrane-based microfluidic device for controlling the flux of platelet agonists into flowing blood. , 2008, Lab on a chip.

[7]  Caterina Minelli,et al.  A micro-fluidic study of whole blood behaviour on PMMA topographical nanostructures , 2008, Journal of nanobiotechnology.

[8]  S. Diamond,et al.  Microfluidic focal thrombosis model for measuring murine platelet deposition and stability: PAR4 signaling enhances shear‐resistance of platelet aggregates , 2008, Journal of thrombosis and haemostasis : JTH.

[9]  Shaun P Jackson,et al.  The growing complexity of platelet aggregation. , 2007, Blood.

[10]  S. Jackson,et al.  Techniques to examine platelet adhesive interactions under flow. , 2004, Methods in molecular biology.

[11]  J. Moake,et al.  Platelets and shear stress. , 1996, Blood.

[12]  Carolyn G. Conant,et al.  New Device for High-Throughput Viability Screening of Flow Biofilms , 2010, Applied and Environmental Microbiology.

[13]  Takashi Nakamura,et al.  Activation of the GP IIb-IIIa Complex Induced by Platelet Adhesion to Collagen Is Mediated by Both α2β1 Integrin and GP VI* , 1999, The Journal of Biological Chemistry.