Isolation and characterization of platelet microvesicles

Abstract

In the recent years, Platelet microvesicles (PMVs) are projected as potential diagnostic and prognostic biomarkers of many diseases like cardiovascular diseases, diabetes and infection. Due to the small size (100-1000 nm), diverse structure and functions, PMVs is associated with different challenges during isolation, quantification and characterization. Hence, it is pertinent to optimize methods to isolate, characterize and quantify MVs in blood plasma. Therefore, in this study it is aimed to isolate and characterize the PMVs release upon platelet activation. The released PMVs were isolated by differential centrifugation and characterized through scanning electron microscopy (SEM) and flow cytometer (FACS). SEM images showed resting platelets as smooth spherical cells of 2- 3 ?m in diameter separated from each other having presence of small hyaloplasmic extensions. In contrast, platelet activation elicits significant morphological change exhibited well-developed pseudopods (hyaloplasmic processes), which connected the cells forming large aggregates (clumps). Electron microscopy and flow cytometry data showed platelet microvesicles release of size in the range of 200-500 nm from the activated platelets. Thus, in this study an isolation and characterization technique for platelet microvesicles was optimized that can be further validated with clinical samples and exploit to study microvesicles structure, function and clinical importance.