Monitoring modulators of platelet aggregation in a microtiter plate assay

Moran, Niamh; Kiernan, Aoife; Dunne, Eimear; Edwards, Richard J.; Shields, Denis C.; Kenny, Dermot

doi:10.1016/j.ab.2006.06.037

Cited by 38 publications

(25 citation statements)

References 28 publications

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“…All reports are agreed on reading between 575 and 650 nm for PRP (9–11,15–17,20,23) which minimized absorption of plasma, and 405 nm for washed platelets (9,21). …”

Section: -Well Plate Aggregationmentioning

confidence: 92%

“…These shakers come in different varieties, e.g. linear (8,16,23), orbital (17,21) and kinetic (10,11,15). In LTA, a stir bar is present in each sample which creates a simple reproducible vortex in the cuvette whereas the internal shakers in plate readers are hugely variable.…”

Section: -Well Plate Aggregationmentioning

confidence: 99%

“…The majority of this work concerns the characterization of platelet reactivity in the presence of antiplatelet therapies such as GPIIb/IIIa inhibitors (16,23) or dual anti-platelet therapy with ADP P2Y 12 antagonists plus aspirin (10–13,17,21,22,25,26). Using these techniques, it is possible to produce concentration-response curves which can be used to evaluate the pharmacological action of these therapies.…”

Section: -Well Plate Aggregationmentioning

confidence: 99%

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96-well plate-based aggregometry

2018

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While there are many bench and bedside tests to assess platelet reactivity, ex vivo light transmission aggregometry (LTA) remains the gold standard. LTA, however, is expensive, time-consuming and requires dedicated equipment and staff, making it impractical in many situations. In addition, there is significant variability between data generated at different testing sites meaning that tests often need to be repeated if a patient is transferred to the care of a different hospital. As such, there is clearly an unmet need for standardization of platelet testing.Using the principles of LTA, aggregometry can be conducted in 96-well plates with readings being made in a standard plate reader. This approach allows for the assessment of multiple concentrations of agonists, since the volume of platelets required for each test is significantly lower than for LTA. Furthermore, the lyophilization of a set panel of agonists to a 96-well plate to produce a stable assay substrate allows the production of portable, standardized plates that can be used to generate reproducible tests at multiple sites.In this review, we will discuss the methods and uses of 96-well plate aggregometry for both research and the clinic.

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“…All reports are agreed on reading between 575 and 650 nm for PRP (9–11,15–17,20,23) which minimized absorption of plasma, and 405 nm for washed platelets (9,21). …”

Section: -Well Plate Aggregationmentioning

confidence: 92%

Section: -Well Plate Aggregationmentioning

confidence: 99%

Section: -Well Plate Aggregationmentioning

confidence: 99%

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96-well plate-based aggregometry

2018

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“…[24][25][26] Washed platelets were resuspended at concentration of 10 8 cells/mL in buffer A, and 1.8 mM CaCl 2 added just prior to assay. 24) Aliquots (100 µL) of platelets dispensed into 96-well microplate were pre-incubated at 37°C for 10 min in the presence of 50 µL of drugs to be tested (first dissolved in buffer A and then filtered through a 0.22 m syringe filter) or buffer A as vehicle, and then were stimulated with 50 µL of agonist.…”

Section: Methodsmentioning

confidence: 99%

Delineation of Platelet Activation Pathway of Scutellarein Revealed Its Intracellular Target as Protein Kinase C

Tian

Chang

et al. 2016

Biological & Pharmaceutical Bulletin

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Erigeron breviscapus has been widely used in traditional Chinese medicine (TCM) and its total flavonoid component is commonly used to treat ischemic stroke, coronary heart disease, diabetes and hypertension. Scutellarin is the major ingredient of E. breviscapus and scutellarein is one of the main bioactive metabolites of scutellarin in vivo, but the latter's pharmacological activities have not been fully characterized. Provided evidence that could inhibit platelet aggregation, the effect of scutellarein on rat washed platelets and its underlying mechanisms were evaluated in our research. Scutellarein inhibited platelet adhesion and aggregation induced by multiple G protein coupled receptor agonists such as thrombin, U 46619 and ADP, in a concentration-dependent manner. Furthermore, the mild effect of scutellarein on intracellular Ca 2 mobilization and cyclic AMP (cAMP) level was observed. On the other hand, the role of scutellarein as potential protein kinase C (PKC) inhibitor was confirmed by PKC activity analysis and molecular docking. The phorbol myristate acetate-induced platelets aggregation assay with or without ADP implied that the scutellarein takes PKC(s) as its primary target(s), and acts on it in a reversible way. Finally, scutellarein as a promising agent exhibited a high inhibition effect on ADP-induced platelet aggregation among its analogues. This study clarifies the PKC-related signaling pathway involved in antiplatelet action of scutellarein, and may be beneficial for the treatment of cardiovascular diseases.

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“…Platelet Aggregation Inhibitors: Platelets play a critical role in haemostasis and the development of cardiac disorder 33 . Platelet hyperactivity is responsible for morphologic changes in the platelets and the release of chemical mediators, such as adenosine diphosphate (ADP), thromboxane A 2 , serotonin, platelet activation factor (PAF) and thrombin and therefore, plays an important role in arterial thrombosis and atherosclerosis.…”

Section: Phytoconstituents and Disorder Typementioning

confidence: 99%

Untitled

2013

IJPSR

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Worldwide, cardiac disorders constitute a leading cause of morbidity and mortality. Cardiac disorders include high blood pressure, atherosclerosis, arrhythmia, congestive heart failure, stroke etc. Increase in incidence of cardiac disorders is a manifestation of lifestyle changes. Although modern drugs are effective in preventing the cardiac disorders, their use is often limited because of their side effects and adverse reactions. Use of herbal drugs is not only cost effective but also has better safety and efficacy. The World health Organisation (WHO) estimates that about 80% of the population living in the developing countries still relies on herbal drugs for their primary healthcare needs. Today, ethno-botanical and ethno-pharmacological studies of medicinal plants continue to attract investigators for research work globally. Aim of this current review to explain herbal phytoconstituents therapy on the basis--Type of disorder, Mode of action and Pharmacological screening model‖, which could be an informatics approach in management of cardiac disorders.

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Monitoring modulators of platelet aggregation in a microtiter plate assay

Cited by 38 publications

References 28 publications

96-well plate-based aggregometry

96-well plate-based aggregometry

Delineation of Platelet Activation Pathway of Scutellarein Revealed Its Intracellular Target as Protein Kinase C

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