A transmission electron microscopy study of anticoagulant-induced platelet vesiculation

Almhanawi, Bahaa Hadi; Khalid, Bahariah; Ibrahim, Tengku Azmi Tengku

doi:10.1016/j.pbj.2016.11.002

Cited by 12 publications

(6 citation statements)

References 31 publications

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“…Furthermore, platelets stored at 4 °C display damage and protrusion of cell membrane, resulting in release of granules and cell organelles (Fig. 4 ) [ 21 ]. Both a longer storage duration and refrigeration activated and influenced the shape and structure of the platelet surface.…”

Section: Resultsmentioning

confidence: 99%

Platelet storage induces accelerated desialylation of platelets and increases hepatic thrombopoietin production

et al. 2018

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BackgroundStored platelets undergo deleterious changes, referred to as platelet storage lesions (PSLs), which accelerate the desialylation of platelets and result in their phagocytosis and clearance by hepatic macrophages. Recent studies have reported that Ashwell–Morell receptor binds to desialylated platelets, thereby inducing hepatic thrombopoietin (TPO) production in a mouse model. Therefore, this study aimed to demonstrate these relationships between PSL and hepatic TPO production in human study.MethodsPlatelet concentrates (PCs) were obtained from 5 healthy volunteers and the remaining were discarded samples from the blood bank. PCs were divided into two halves, and stored either at 22 or 4 °C. Experiments were conducted using serial samples. Desialylation was assessed using flow cytometry, and structural changes were visualized using electron microscopy. Following co-culture of HepG2 cells (HB-8065, ATCC) with isolated platelets, hepatic TPO production was determined using real-time quantitative polymerase chain reaction and the supernatant TPO level was measured using a Luminex kit.ResultsFor 5 days of storage duration, platelet counts were not influenced by the storage conditions, but the degree of desialylation was proportional to the storage duration. Significant changes in the platelet surface and structure according to storage conditions were noted in electron microscopy. HepG2 cells incubated with aged platelets expressed more TPO mRNA, and supernatant TPO levels were proportional to the storage duration. Refrigeration also influenced on the results of this study, but they were not statistically significant.ConclusionsThis is the first study to demonstrate that, in vitro, aging and refrigeration affect the integrity of human platelets, resulting in induction of hepatic TPO mRNA and protein expression.

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Section: Resultsmentioning

confidence: 99%

Platelet storage induces accelerated desialylation of platelets and increases hepatic thrombopoietin production

et al. 2018

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“…Full of cytoplasmic vacuoles and phagosomes containing organic (cellular, microbial) as well as inorganic foreign materials and have a leading pseudopodium in one direction [85,86] Dendritic cell Show a highly vacuolated appearance and large number of long dendritic processes to give them a very large surface-to-volume ratio for antigen presentation [87][88][89] Platelet Platelet Dense granule membrane proteins incorporate with the platelet plasma membrane, formation of small cell protrusions, cytoskeletal proteins rearrangement leading to more ameboid shape [73] Please note that macrophages and mast cells are mostly tissue-resident cells and less likely to be found in periphery [90,91].…”

Section: Macrophagementioning

confidence: 99%

“…PlateletPlatelet 5-3 μm in diameter, with no nucleus, but contains multiple vesicles and granules[73] Granulocytes Basophil8-11μm in diameter and contain large secretory granules and lipid bodies [74, 75] Eosinophil 11-14 μm in diameter, often have a bilobed nucleus and contain numerous distinctive ellipsoid granules with a linear electron dense crystalline core [62, 76] Neutrophil 9-12 μm in diameter, contain small granules of various types and a lobulated nucleus [75, 77] Mast cell 8-20 μm in diameter contain large numbers of cytoplasmic granules that are smaller in size than those of basophils. Mast cell surface tends to have narrow elongated protrusions [78, 79] Stimulated PBMCs T cell Activated cells have rougher cell membrane with relatively little rough ER and filled with free ribosomes [69, 80,81]B cell Activated cells have abundant cytoplasm filled with an extensive rough endoplasmic reticulum (ER)[69,82] …”

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confidence: 99%

Phenotypic characteristics of peripheral immune cells of Myalgic encephalomyelitis/chronic fatigue syndrome via transmission electron microscopy: A pilot study

et al. 2022

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Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a complex chronic multi-systemic disease characterized by extreme fatigue that is not improved by rest, and worsens after exertion, whether physical or mental. Previous studies have shown ME/CFS-associated alterations in the immune system and mitochondria. We used transmission electron microscopy (TEM) to investigate the morphology and ultrastructure of unstimulated and stimulated ME/CFS immune cells and their intracellular organelles, including mitochondria. PBMCs from four participants were studied: a pair of identical twins discordant for moderate ME/CFS, as well as two age- and gender- matched unrelated subjects—one with an extremely severe form of ME/CFS and the other healthy. TEM analysis of CD3/CD28-stimulated T cells suggested a significant increase in the levels of apoptotic and necrotic cell death in T cells from ME/CFS patients (over 2-fold). Stimulated Tcells of ME/CFS patients also had higher numbers of swollen mitochondria. We also found a large increase in intracellular giant lipid droplet-like organelles in the stimulated PBMCs from the extremely severe ME/CFS patient potentially indicative of a lipid storage disorder. Lastly, we observed a slight increase in platelet aggregation in stimulated cells, suggestive of a possible role of platelet activity in ME/CFS pathophysiology and disease severity. These results indicate extensive morphological alterations in the cellular and mitochondrial phenotypes of ME/CFS patients’ immune cells and suggest new insights into ME/CFS biology.

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“…3 In studies evaluating the ultrastructures of platelets by electron microscopic analysis, this chemical exposure to EDTA is associated with platelet swelling and increases in diameter. 5 Lance et al reported that the optimal measurement time for MPV should be 60 minutes or 120 minutes after blood collection, respectively, depending on whether citrate or EDTA is used as the anticoagulant. 6 Various studies have reported that when using EDTA as an anticoagulant, MPV values might be deviated by 2-50% depending upon the measurement time.…”

Section: Sirmentioning

confidence: 99%

Mean platelet volume may not be a marker for disease activity in children with systemic lupus erythematosus

Beyan

2021

Lupus

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A transmission electron microscopy study of anticoagulant-induced platelet vesiculation

Cited by 12 publications

References 31 publications

Platelet storage induces accelerated desialylation of platelets and increases hepatic thrombopoietin production

Platelet storage induces accelerated desialylation of platelets and increases hepatic thrombopoietin production

Phenotypic characteristics of peripheral immune cells of Myalgic encephalomyelitis/chronic fatigue syndrome via transmission electron microscopy: A pilot study

Mean platelet volume may not be a marker for disease activity in children with systemic lupus erythematosus

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