Electric cell-substrate impedance sensing (ECIS) is a quickly advancing field to measure the barrier function of endothelial cells. Most ECIS systems that are commercially available use gold electrodes, which are opaque and do not allow for real-time imaging of cellular responses. In addition, most ECIS systems have a traditional tissue culture Petri-dish set up. This conventional set-up does not allow the introduction of physiologically relevant shear stress, which is crucial for the endothelial cell barrier function. Here, we created a new ECIS micro-bioreactor (MBR) that incorporates a clear electrode made of indium tin oxide in a microfluidic device. Using this device, we demonstrate the ability to monitor the barrier function along culture of cells under varying flow rates. We show that while two cell types align in the direction of flow in responses to high shear stress, they differ in the barrier function. Additionally, we observe a change in the barrier function in response to chemical perturbation. Following exposure to EDTA that disrupts cell-to-cell junctions, we could not observe distinct morphological changes but measured a loss of impedance that could be recovered with EDTA washout. High magnification imaging further demonstrates the loss and recovery of the barrier structure. Overall, we establish an ECIS MBR capable of real-time monitoring of the barrier function and cell morphology under shear stress and allowing high-resolution analysis of the barrier structure.
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July 2018
Research Article|
May 15 2018
Microfluidic platform for the real time measurement and observation of endothelial barrier function under shear stress
Special Collection:
Bio-transport processes and drug delivery in physiological micro-devices
Daniel M. Lewis;
Daniel M. Lewis
a)
1
Department of Chemical and Biomolecular Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
2
Johns Hopkins Physical Sciences–Oncology Center and Institute for NanoBioTechnology, Johns Hopkins University
, Baltimore, Maryland 21218, USA
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Nicholas Mavrogiannis;
Nicholas Mavrogiannis
a)
1
Department of Chemical and Biomolecular Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
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Zachary Gagnon;
Zachary Gagnon
1
Department of Chemical and Biomolecular Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
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Sharon Gerecht
Sharon Gerecht
b)
1
Department of Chemical and Biomolecular Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
2
Johns Hopkins Physical Sciences–Oncology Center and Institute for NanoBioTechnology, Johns Hopkins University
, Baltimore, Maryland 21218, USA
3
Department of Materials Science and Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
b)Author to whom correspondence should be addressed: [email protected]. Tel.: +1-410-516-2846. Fax: +1-410-516-5510.
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Daniel M. Lewis
1,2,a)
Nicholas Mavrogiannis
1,a)
Zachary Gagnon
1
Sharon Gerecht
1,2,3,b)
1
Department of Chemical and Biomolecular Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
2
Johns Hopkins Physical Sciences–Oncology Center and Institute for NanoBioTechnology, Johns Hopkins University
, Baltimore, Maryland 21218, USA
3
Department of Materials Science and Engineering, Johns Hopkins University
, Baltimore, Maryland 21218, USA
a)
D. M. Lewis and N. Mavrogiannis contributed equally to this work.
b)Author to whom correspondence should be addressed: [email protected]. Tel.: +1-410-516-2846. Fax: +1-410-516-5510.
Biomicrofluidics 12, 042202 (2018)
Article history
Received:
February 25 2018
Accepted:
April 10 2018
Citation
Daniel M. Lewis, Nicholas Mavrogiannis, Zachary Gagnon, Sharon Gerecht; Microfluidic platform for the real time measurement and observation of endothelial barrier function under shear stress. Biomicrofluidics 1 July 2018; 12 (4): 042202. https://doi.org/10.1063/1.5026901
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