Storage conditions determine the characteristics of red blood cell derived extracellular vesicles

Abstract

Extracellular vesicles (EVs) are released during the storage of red blood cell (RBC) concentrates and might play adverse or beneficial roles throughout the utilization of blood products (transfusion). Knowledge of EV release associated factors and mechanism amends blood product management. In the present work the impact of storage time and medium (blood preserving additive vs isotonic phosphate buffer) on the composition, size, and concentration of EVs was studied using attenuated total reflection infrared (ATR-IR) spectroscopy, microfluidic resistive pulse sensing (MRPS) and freeze-fraction combined transmission electron micrography (FF-TEM). The spectroscopic protein-to-lipid ratio based on amide and the C–H stretching band intensity ratio indicated the formation of various vesicle subpopulations depending on storage conditions. After short storage, nanoparticles with high relative protein content were detected. Spectral analysis also suggested differences in lipid and protein composition, too. The fingerprint region (from 1300 to 1000 cm−1) of the IR spectra furnishes additional information about the biomolecular composition of RBC-derived EVs (REVs) such as adenosine triphosphate (ATP), lactose, glucose, and oxidized hemoglobin. The difference between the vesicle subpopulations reveals the complexity of the REV formation mechanism. IR spectroscopy, as a quick, cost-effective, and label-free technique provides valuable novel biochemical insight and might be used complementary to traditional omics approaches on EVs.

Extracellular Vesicles From Stored Red Blood Cells Convey Heme and Induce Spic Expression on Human Monocytes

“…According to the literature, the storage of blood at 4°C leads to RBC injury even in the presence of blood stabilizers, with hemolysis and the release of RBC intracellular contents ( 27 ) and RBC derived EVs ( 28 – 30 ) into the bloodstream. Corroborating these data, the results presented herein demonstrated increased free heme levels starting from day 15, further augmenting daily until day 21 ( Figure 2A ).…”

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

“…Despite a previously documented lower influence of EDTA at lower PEV concentrations, it has the best impact on PEVs stability during storage. It was documented that during the storage of whole blood or platelet concentrates, the number of erythrocyte-derived EVs and PEVs increases [ 43 , 44 ]. In another study, the PEV count in EDTA was stable for 48 h [ 45 ].…”

Blood Cell-Derived Microvesicles in Hematological Diseases and beyond

“…Studied as a separate group of MVs, due to the completely different cellular and plasma environment, low temperature, and absence of clearance mechanisms, microvesicles derived from RBCs stored under blood bank conditions (sRMVs) also show some interesting features [ 185 ]. First of all, they are accumulated exponentially in the supernatant of the RBC units by the storage time and their composition seems to follow the in vitro aging of the parent cells, whereas these characteristics are also affected by the specific storage conditions, for example the storage medium [ 83 , 186 ]. Moreover, they are characterized by increased fibrinolytic activity due to the presence of plasminogen on their surface (that decreases, however, by the storage time), as well as to antithrombin activity, probably related with the presence of a2-macroglobulin [ 187 , 188 ].…”