Single-Step Separation of Platelets from Whole Blood Coupled with Digital Quantification by Interfacial Platelet Cytometry (iPC)

Basabe‐Desmonts, Lourdes; Ramström, Sofia; Meade, Gerardene; O‘Neill, Sarah; Riaz, Asif; Lee, L. P.; Ricco, Antonio J.; Kenny, Dermot

doi:10.1021/la9039682

Cited by 43 publications

(51 citation statements)

References 22 publications

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“…[20][21][22][23] In this work, we apply microcontact printing to create micro/nanoscale patterns of matrix proteins that mediate platelet adhesion and activation, to demonstrate that the microenvironmental geometry directly mediates platelet physiology and function. Specifically, we demonstrate that platelet a-granule secretion is spatially regulated at the micro/nanoscale via rearrangement of the actin cytoskeleton.…”

Section: -11mentioning

confidence: 99%

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Sakurai

Fitch-Tewfik

Qiu

et al. 2015

Blood

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• The geometric orientation of the underlying matrix regulates platelet a-granule secretion.• On geometrically constrained matrices, platelets selfdeposit additional matrix, providing more cell membrane to extend spreading.Although the biology of platelet adhesion on subendothelial matrix after vascular injury is well characterized, how the matrix biophysical properties affect platelet physiology is unknown. Here we demonstrate that geometric orientation of the matrix itself regulates platelet a-granule secretion, a key component of platelet activation. Using protein microcontact printing, we show that platelets spread beyond the geometric constraints of fibrinogen or collagen micropatterns with <5-mm features. Interestingly, a-granule exocytosis and deposition of the a-granule contents such as fibrinogen and fibronectin were primarily observed in those areas of platelet extension beyond the matrix protein micropatterns. This enables platelets to "self-deposit" additional matrix, provide more cellular membrane to extend spreading, and reinforce platelet-platelet connections. Mechanistically, this phenomenon is mediated by actin polymerization, Rac1 activation, and a IIb b 3 integrin redistribution and activation, and is attenuated in gray platelet syndrome platelets, which lack a-granules, and Wiskott-Aldrich syndrome platelets, which have cytoskeletal defects. Overall, these studies demonstrate how platelets transduce geometric cues of the underlying matrix geometry into intracellular signals to extend spreading, which endows platelets spatial flexibility when spreading onto small sites of exposed subendothelium. (Blood. 2015;126(4):531-538) IntroductionHemostasis begins with platelet adhesion and activation at the site of vascular injury.1 Platelets then secrete their a-and dense granules, leading to further recruitment of other platelets and the formation of a hemostatic plug.2 Extensive research has characterized the underlying biological signaling pathways that govern these processes at the bulk level via in vivo and in vitro techniques.3 However, how platelets interact with their microenvironment at the single-cell level and how these interactions subsequently influence platelet physiology remain poorly understood. For example, although single platelets have long been observed to fill submicron-sized "gaps" in the endothelium, 4 how platelets establish adhesion on such small areas of exposed matrix that is strong enough to withstand the shear forces of the circulation is unknown. Studies of single platelet-matrix interactions may help explain the role of platelets in maintaining vascular integrity and supporting the semipermeable barrier function of the endothelium during homeostatic conditions. 5 Furthermore, because platelets within different regions of the same thrombus exhibit significantly different levels of activation, 6,7 this concept of physical and microenvironmental regulation of platelet physiology at the single-cell level is also important for our overall understanding of clot formation. S...

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Section: -11mentioning

confidence: 99%

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Sakurai

Fitch-Tewfik

Qiu

et al. 2015

Blood

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“…A 3D PDMS microfluidic chip with an integrated array of microwells capable of long-term tumor spheroid cultivation and the evaluation of anticancer drug activity has been developed [312]. By culturing spheroids of HT-29 human carcinoma cells onto a microfluidic chip over the course of four weeks, introducing a cytostatic drug (5-fluorouracil), and incubating on the PDMS chip, the cytotoxic effect on HT-29 cells could be evaluated.…”

Section: Cell Analysismentioning

confidence: 99%

Polymeric-Based In Vitro Diagnostic Devices

Cheng

Kuan

Chen

2015

In-Vitro Diagnostic Devices

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“…Platelets are the smallest type of cells found in blood (~2-3 µm diameter), and they are involved in many fundamental physiological processes such as hemostasis and thrombosis [62]. When bleeding from a wound occurs, circulating platelets adhere and aggregate at the injury site to initiate thrombus formation until the bleeding is stopped.…”

Section: Platelet Separationmentioning

confidence: 99%

“…Recently, single-step separation of individual platelets from whole blood by micropatterning of platelet-specific protein surfaces on solid substrates has been reported using an interfacial platelet cytometry (iPC) chip [62]. Platelets were separated from whole blood using iPC coated with fibrinogen, von Willebrand factor (VWF), and anti-CD42b antibody printed "spots" ranging from 2-24 μm diameter.…”

Section: Platelet Separationmentioning

confidence: 99%

Microfluidic Devices for Blood Fractionation

et al. 2011

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Blood, a complex biological fluid, comprises 45% cellular components suspended in protein rich plasma. These different hematologic components perform distinct functions in vivo and thus the ability to efficiently fractionate blood into its individual components has innumerable applications in both clinical diagnosis and biological research. Yet, processing blood is not trivial. In the past decade, a flurry of new microfluidic based technologies has emerged to address this compelling problem. Microfluidics is an attractive solution for this application leveraging its numerous advantages to process clinical blood samples. This paper reviews the various microfluidic approaches realized to successfully fractionate one or more blood components. Techniques to separate plasma from hematologic cellular components as well as isolating blood cells of interest including certain rare cells are discussed. Comparisons based on common separation metrics including efficiency (sensitivity), purity (selectivity), and OPEN ACCESSMicromachines 2011, 2 320 throughput will be presented. Finally, we will provide insights into the challenges associated with blood-based separation systems towards realizing true point-of-care (POC) devices and provide future perspectives.

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Single-Step Separation of Platelets from Whole Blood Coupled with Digital Quantification by Interfacial Platelet Cytometry (iPC)

Cited by 43 publications

References 22 publications

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Platelet geometry sensing spatially regulates α-granule secretion to enable matrix self-deposition

Polymeric-Based In Vitro Diagnostic Devices

Microfluidic Devices for Blood Fractionation

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