Extraction and identification of platelet‑derived microparticles

Abstract

Microparticles are carriers of signals for intracellular signal transduction. These carriers include proteins, mrnas, micrornas and other bioactive substances. Platelets are a major source of circulating microparticles, and microparticles are closely associated with the development of certain cardiovascular diseases. in the present study, a method for separating, extracting and identifying platelet-derived microparticles was developed and differences in the expression of surface proteins on microparticles harvested from platelets stimulated by vortexing or treatment with thrombin was investigated. The counts, composition, sizes and inner structures of microparticles were determined using flow cytometry and transmission electron microscopy. additionally, it was demonstrated that platelets could be readily activated, and a large quantity of microparticles with varying complex compositions, structures and sizes were derived from activated platelets. High purity platelet-derived microparticles were obtained by gradient centrifugation. However, the microparticles derived from platelets stimulated by thrombin treatment or vortexing differed significantly in the levels of CD63. The present study aimed to provide improved options for the extraction and identification of microparticles.

Comparison and Investigation of Exosomes Derived from Platelet-Rich Plasma Activated by Different Agonists

“…When being used in clinical applications, methods of activating PRP to release exosomes should be rapid and standardized, produce a high yield of pure exosomes from small PRP volumes, and be compatible with a large number of samples. A large number of comparative studies have been performed, which compare different activators to activate platelets to release exosomes 20 , 27 , 28 , 30 , 31 . However, these activation steps are cumbersome.…”

Platelet Extracellular Vesicles Are Taken up by Canine T Lymphocytes but Do Not Play a Role in Their Proliferation, Differentiation and Cytokine Production In Vitro

“…In our study, the PEVs were incorporated only by T lymphocytes, thus, we evaluated only PEVs influence on the cells. The T cells uptake PEVs within 7 h, but not after 20 h. We consider that PEVs with their expression of phosphatidylserine were effectively taken up an phagocyted by monocytes in cell culture, which was observed by Weiss et al In contrast, in Sadallah’ work, PEVs were detected on CD4 + T cells after 20 h of incubation only CD4+ and CD8 + T cells subsets [ 49 , 52 ].…”

“…Moreover, several studies have been performed, comparing the effect of different PLT activators on differences in PEV release [ 49 ]. Aatonen and her colleagues showed a list of diminishing PLT activators: Ca++ ionophore > thrombin > CRP-XL > TC co-stimulus > collagen > LPS > TRAP-6 > ADP [ 50 ].…”

Characterization and Therapeutic Use of Extracellular Vesicles Derived from Platelets

“…Different processes of platelet activation may lead to the formation of heterogeneous PL-EV populations with different surface marker expression and protein profiles, which may affect their role in intercellular communication [ 99 ]. Several comparative studies have been performed, which compared the effect of different activators on platelet activation and the subsequent release of exosomes [ 49 , 95 , 100 , 101 ]. For example, EVs from platelets activated in vitro with ADP contain different protein cargo in comparison with those activated by collagen or collagen and thrombin [ 99 ], implying that these EVs could also have different functions.…”