In vitro modeling of the microvascular occlusion and thrombosis that occur in hematologic diseases using microfluidic technology
Michelle Tsai, … , Daniel A. Fletcher, Wilbur A. Lam
Michelle Tsai, … , Daniel A. Fletcher, Wilbur A. Lam
Published January 3, 2012; First published December 12, 2011
Citation Information: J Clin Invest. 2012;122(1):408-418. https://doi.org/10.1172/JCI58753.
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Technical Advance Hematology

In vitro modeling of the microvascular occlusion and thrombosis that occur in hematologic diseases using microfluidic technology

Abstract

In hematologic diseases, such as sickle cell disease (SCD) and hemolytic uremic syndrome (HUS), pathological biophysical interactions among blood cells, endothelial cells, and soluble factors lead to microvascular occlusion and thrombosis. Here, we report an in vitro “endothelialized” microfluidic microvasculature model that recapitulates and integrates this ensemble of pathophysiological processes. Under controlled flow conditions, the model enabled quantitative investigation of how biophysical alterations in hematologic disease collectively lead to microvascular occlusion and thrombosis. Using blood samples from patients with SCD, we investigated how the drug hydroxyurea quantitatively affects microvascular obstruction in SCD, an unresolved issue pivotal to understanding its clinical efficacy in such patients. In addition, we demonstrated that our microsystem can function as an in vitro model of HUS and showed that shear stress influences microvascular thrombosis/obstruction and the efficacy of the drug eptifibatide, which decreases platelet aggregation, in the context of HUS. These experiments establish the versatility and clinical relevance of our microvasculature-on-a-chip model as a biophysical assay of hematologic pathophysiology as well as a drug discovery platform.

Authors

Michelle Tsai, Ashley Kita, Joseph Leach, Ross Rounsevell, James N. Huang, Joel Moake, Russell E. Ware, Daniel A. Fletcher, Wilbur A. Lam

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Figure 1

An in vitro microfluidic model of the microvasculature for investigating disease processes involving biophysical cellular interactions.

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An in vitro microfluidic model of the microvasculature for investigating...
(A) Macroscopic view of PDMS microdevice. (B) Software-generated image used to develop the photolithography mask that defines the geometric pattern of the microfluidic channels. Smallest channels in this pattern are 30 μm wide. Scale bar: 600 μm. (C) Brightfield images show that within 48 hours, HUVECs seeded into the microdevice are cultured to confluency. Images are taken from different areas of the same device at the same scale. Scale bars: 30 μm. (D) 3D renderings of multiple confocal microscopy using fluorescent cell membrane (red) and cell nuclear (blue) dyes show that the endothelial cells line the entire inner surface of the microfluidic channels. The cross-sectional view also reveals that the cells round off the square corners of the smallest microchannels. Scale bars: 30 μm.