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Procoagulant tumour microvesicles attach to endothelial cells on biochips under microfluidic flow

Algarni, Abdulrahman; Greenman, John; Madden, Leigh

Authors

Abdulrahman Algarni



Abstract

Tumor patients are at a high risk of venous thromboembolism (VTE), and the mechanism by which this occurs may involve tumor-derived microvesicles (MVs). Previously, it has been shown that tumor MVs become attached to endothelial cells in static conditions. To investigate whether this process occurs under physiologically relevant flow rates, tumor MVs were perfused across a microfluidic device coated with growing human umbilical vein endothelial cells (HUVECs). Cell lines were screened for their ability to form tumor spheroids, and two cell lines, ES-2 and U87, were selected; spheroids formed were transferred to a microfluidic chip, and a second endothelial cell biochip was coated with HUVECs and the two chips were linked. Media flowed through the spheroid chip to the endothelial chip, and procoagulant activity (PCA) of the tumor media was determined by a one-stage prothrombin time assay. Tumor MVs were also quantified by flow cytometry before and after interaction with HUVECs. Confocal images showed that HUVECs acquired fluorescence from MV attachment. Labeled MVs were proportionally lost from MV rich media with time when flowed over HUVECs and were not observed on a control chip. The loss of MV was accompanied by a proportional reduction in PCA. Flow cytometry, confocal microscopy, and live flow imagery captured under pulsatile flow confirmed an association between tumor MVs and HUVECs. Tumor MVs attached to endothelial cells under physiological flow rates, which may be relevant to the VTE pathways in cancer patients

Citation

Algarni, A., Greenman, J., & Madden, L. (2019). Procoagulant tumour microvesicles attach to endothelial cells on biochips under microfluidic flow. Biomicrofluidics, 13(6), https://doi.org/10.1063/1.5123462

Journal Article Type Article
Acceptance Date Nov 24, 2019
Online Publication Date Dec 6, 2019
Publication Date Dec 6, 2019
Deposit Date Nov 27, 2019
Publicly Available Date Nov 28, 2019
Journal Biomicrofluidics
Electronic ISSN 1932-1058
Publisher AIP Publishing
Peer Reviewed Peer Reviewed
Volume 13
Issue 6
Article Number 064124
DOI https://doi.org/10.1063/1.5123462
Keywords Thrombosis; Microparticles; Microvesicles; Endothelium
Public URL https://hull-repository.worktribe.com/output/3260110

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Copyright Statement
© Authors. This article may be downloaded for personal use only. Any other use requires prior permission of the author and AIP Publishing. The published article appeared in Biomicrofluidics 13, 064124 (2019) and may be found at https://aip.scitation.org/doi/full/10.1063/1.5123462






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