Flow cytometric assays for quantifying activated ovine platelets.
Ovines are a common animal model for preclinical evaluation of cardiovascular devices including heart valves, endovascular grafts, and ventricular assist devices. Biocompatibility is essential to the success of these devices; however, tools to assess biocompatibility in ovines are limited. To address this need, antibodies that bind to activated human and bovine platelets and annexin V protein were evaluated for potential cross-reactivity to activated ovine platelets. These candidate markers were incubated with stimulated and quiescent ovine whole blood, and binding to platelets was quantified by flow cytometry. Several antihuman CD62P antibodies including one polyclonal antibody, three monoclonal antibodies, and annexin V selectively bound to activated ovine platelets. An assay to quantify platelet microaggregates was also developed. The availability of assays to quantify ovine platelet activation can increase the quality of biocompatibility data obtainable during preclinical development of artificial organs in the ovine model, potentially aiding in the evaluation of design refinements to enhance device biocompatibility.
Johnson
Carl A
CA
Bioengineering Department, University of Pittsburgh, PA, USA.
Snyder
Trevor A
TA
Woolley
Joshua R
JR
Wagner
William R
WR
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001187:Artificial Organs; D001672:Biocompatible Materials; D002417:Cattle; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D023421:Models, Animal; D015539:Platelet Activation; D010976:Platelet Count / Q000295:instrumentation / Q000379:methods*; D034561:Sheep, Domestic
D000911:Antibodies, Monoclonal; D001672:Biocompatible Materials