Microfluidic assessment of functional culture-derived platelets in human thrombi under flow

Kamat, Viraj; Muthard, Ryan W.; Li, Ruizhi; Diamond, Scott L.

doi:10.1016/j.exphem.2015.06.302

Cited by 4 publications

(3 citation statements)

References 40 publications

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“…Although the mechanistic underpinnings related to changes in contractile force with various diseases remain unclear, there does appear to be an optimal platelet contractile force associated with healthy hemostasis. 33 Noting the recent developments in creating culture-derived platelets [81][82][83] and interest in improved ways to examine their function, 84 measuring bulk clot and single platelet contraction force may be an important avenue for validating function. Similar validations may be useful for platelets that have undergone storage, especially when transfusing into patients that already have platelets with impaired contractile forces and bleeding.…”

Section: Future Implicationsmentioning

confidence: 99%

Feeling the Force: Measurements of Platelet Contraction and Their Diagnostic Implications

Williams

Oshinowo

Ravindran

et al. 2018

Semin Thromb Hemost

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In addition to the classical biological and biochemical framework, blood clots can also be considered as active biomaterials composed of dynamically contracting platelets, nascent polymeric fibrin that functions as a matrix scaffold, and entrapped blood cells. As platelets sense, rearrange, and apply forces to the surrounding microenvironment, they dramatically change the material properties of the nascent clot, increasing its stiffness by an order of magnitude. Hence, the mechanical properties of blood clots are intricately tied to the forces applied by individual platelets. Research has also shown that the pathophysiological changes in clot mechanical properties are associated with bleeding and clotting disorders, cancer, stroke, ischemic heart disease, and more. By approaching the study of hemostasis and thrombosis from a biophysical and mechanical perspective, important insights have been made into how the mechanics of clotting and the forces applied by platelets are linked to various diseases. This review will familiarize the reader with a mechanics framework that is contextualized with relevant biology. The review also includes a discussion of relevant tools used to study platelet forces either directly or indirectly, and finally, concludes with a summary of potential links between clotting forces and disease.

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Section: Future Implicationsmentioning

confidence: 99%

Feeling the Force: Measurements of Platelet Contraction and Their Diagnostic Implications

Williams

Oshinowo

Ravindran

et al. 2018

Semin Thromb Hemost

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show abstract

“…19,20 Microfluidic flow chambers are currently used to study thrombotic processes by monitoring platelet incorporation into the clot, to study cell adhesion in dynamic conditions or to test the function of blood bank platelets in whole or reconstituted blood. 21,22 This process is recognized as a powerful, promising method to investigate platelet function.…”

Section: Introductionmentioning

confidence: 99%

Functional Validation of microRNA-126-3p as a Platelet Reactivity Regulator Using Human Haematopoietic Stem Cells

et al. 2019

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Background Platelets are an abundant source of micro-ribonucleic acids (miRNAs) that may play a role in the regulation of platelet function. Some miRNAs, such as miR-126-3p, have been noted as potential biomarkers of platelet reactivity and the recurrence of cardiovascular events. However, the biological relevance of these associations remains uncertain, and the functional validation of candidate miRNAs on human-derived cells is lacking. Objective This article functionally validates miR-126-3p as a regulator of platelet reactivity in platelet-like structures (PLS) derived from human haematopoietic stem cells. Materials and Methods CD34+-derived megakaryocytes were transfected with miR-126-3p and differentiated in PLS. PLS reactivity was assessed using perfusion in a fibrinogen-coated flow chamber. miR-126-3p's selected gene targets were validated using quantitative polymerase chain reaction, protein quantification and a reporter gene assay. Results CD34+-derived megakaryocytes transfected with miR-126-3p generated PLS exhibiting 156% more reactivity than the control. These functional data were in line with those obtained analysing CD62P expression. Moreover, miR-126-3p transfection was associated with the down-regulation of a disintegrin and metalloproteinase-9 (ADAM9) messenger RNA (mRNA), a validated target of miR-126-3p, and of Plexin B2 (PLXNB2) mRNA and protein, an actin dynamics regulator. Silencing PLXNB2 led to similar functional results to miR-126-3p transfection. Finally, using a reporter gene assay, we validated PLXNB2 as a direct target of miR-126-3p. Conclusion We functionally validated miR-126-3p as a regulator of platelet reactivity in PLS derived from human haematopoietic stem cells. Moreover, PLXNB2 was validated as a new gene target of miR-126-3p in human cells, suggesting that miR-126-3p mediates its effect on platelets, at least in part, through actin dynamics regulation.

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“…76,82 PLS adhesion can be assessed in static or dynamic conditions since these techniques require only low concentrations of platelets. 58,83 It is noteworthy that PLS may differ substantially from their native counterparts and that, depending on the culture conditions, the reported findings might not replicate the in vivo situation. miRNA-related in vitro findings might also be affected by potential contamination of cell-culture reagents with miRNAs.…”

Section: In Vitro Modelsmentioning

confidence: 91%

Methods to Investigate miRNA Function: Focus on Platelet Reactivity

et al. 2020

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MicroRNAs (miRNAs) are small noncoding RNAs modulating protein production. They are key players in regulation of cell function and are considered as biomarkers in several diseases. The identification of the proteins they regulate, and their impact on cell physiology, may delineate their role as diagnostic or prognostic markers and identify new therapeutic strategies. During the last 3 decades, development of a large panel of techniques has given rise to multiple models dedicated to the study of miRNAs. Since plasma samples are easily accessible, circulating miRNAs can be studied in clinical trials. To quantify miRNAs in numerous plasma samples, the choice of extraction and purification techniques, as well as normalization procedures, are important for comparisons of miRNA levels in populations and over time. Recent advances in bioinformatics provide tools to identify putative miRNAs targets that can then be validated with dedicated assays. In vitro and in vivo approaches aim to functionally validate candidate miRNAs from correlations and to understand their impact on cellular processes. This review describes the advantages and pitfalls of the available techniques for translational research to study miRNAs with a focus on their role in regulating platelet reactivity.

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Microfluidic assessment of functional culture-derived platelets in human thrombi under flow

Cited by 4 publications

References 40 publications

Feeling the Force: Measurements of Platelet Contraction and Their Diagnostic Implications

Feeling the Force: Measurements of Platelet Contraction and Their Diagnostic Implications

Functional Validation of microRNA-126-3p as a Platelet Reactivity Regulator Using Human Haematopoietic Stem Cells

Methods to Investigate miRNA Function: Focus on Platelet Reactivity

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