Evaluation of a Method for Removal and Determination of Thrombogenic Microparticles

• Published in: Blood 2007-11, Vol.110 (11), p.3626-3626
• Main Authors: Vetr, Helga, Graf, Markus, Geiter, Sabine, Oberreither, Manfred, Binder, Bernd R.
• Format: Article
• Language: English
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V110.11.3626.3626

It is now generally accepted that circulating cell-derived microparticles play a major role in thrombotic diseases. Currently available methods to analyse microparticles are not easy to standardize, need specialized technical equipment or detect only subpopulations of microparticles. In this study a new method for quantification of circulating thrombogenic microparticles (MP) is evaluated (Technothrombin® MP Microparticle and Ceveron ® MFU-500). The principle of this method is based on the differences in thrombin generation between platelet poor plasma (PPP) and microparticle free plasma (MPFP) obtained by a standardized filtration meethod. This difference in thrombin generation between PPP and MPFP plasma reflects the amount of microparticles contained in PPP and removed by filtration. To evaluate this method, PPP from normal blood donors was prepared by centrifugation for 15 min at 2,500xg. MPFP was generated by filtration (Ceveron® MFU-500; 0.2 μm). For comparison MPFP was also prepared by high speed centrifugation (15,000xg for 30 min). All samples were analysed for thrombin generation using the Technothrombin®TGA method. For calibration purposes, dilutions of purified MP from red blood cells were prepared in MPFP plasma and thrombin generation was measured before and after filtration of each dilution. Recovery of MP from the filter membrane was performed by rinsing the membrane with an equal volume of standard MP free plasma. In addition, filtered and non-filtered samples were analyzed in standard coagulation assays (PT, aPTT, Fibrinogen, FVIII activity, Lupus). Peak thrombin values from centrifuged (57nM±8) or filtered samples (79nM±11) were not significantly different from each other (p=0.14) but were significantly lower (p