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Real-Time Measurement of Melanoma Cell-Mediated Human Brain Endothelial Barrier Disruption Using Electric Cell-Substrate Impedance Sensing Technology

1
Department of Molecular Medicine and Pathology, School of Medical Sciences, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1023, New Zealand
2
Centre for Brain Research, University of Auckland, Auckland 1023, New Zealand
3
Auckland Cancer Society Research Centre, University of Auckland, Auckland 1023, New Zealand
4
School of Biological Sciences, Faculty of Science, University of Auckland, Auckland 1010, New Zealand
5
Department of Anatomy and Medical Imaging, School of Medical Sciences, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1023, New Zealand
6
Maurice Wilkins Centre for Molecular Biodiscovery, University of Auckland, Auckland 1010, New Zealand
*
Author to whom correspondence should be addressed.
Biosensors 2019, 9(2), 56; https://doi.org/10.3390/bios9020056
Received: 25 February 2019 / Revised: 26 March 2019 / Accepted: 9 April 2019 / Published: 15 April 2019
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Abstract

Electric cell-substrate impedance sensing (ECIS) is an impedance-based method for monitoring changes in cell behaviour in real-time. In this paper, we highlight the importance of ECIS in measuring the kinetics of human melanoma cell invasion across human brain endothelium. ECIS data can be mathematically modelled to assess which component of the endothelial paracellular and basolateral barriers is being affected and when. Our results reveal that a range of human melanoma cells can mediate disruption of human brain endothelium, primarily involving the paracellular route, as demonstrated by ECIS. The sensitivity of ECIS also reveals that the paracellular barrier weakens within 30–60 min of the melanoma cells being added to the apical face of the endothelial cells. Imaging reveals pronounced localisation of the melanoma cells at the paracellular junctions consistent with paracellular migration. Time-lapse imaging further reveals junctional opening and disruption of the endothelial monolayer by the invasive melanoma cells all within several hours. We suggest that the ability of ECIS to resolve changes to barrier integrity in real time, and to determine the route of migration, provides a powerful tool for future studies investigating the key molecules involved in the invasive process of cancer cells. View Full-Text
Keywords: ECIS; impedance; barrier function; endothelium; blood–brain barrier ECIS; impedance; barrier function; endothelium; blood–brain barrier
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Anchan, A.; Kalogirou-Baldwin, P.; Johnson, R.; Kho, D.T.; Joseph, W.; Hucklesby, J.; Finlay, G.J.; O’Carroll, S.J.; Angel, C.E.; Graham, E.S. Real-Time Measurement of Melanoma Cell-Mediated Human Brain Endothelial Barrier Disruption Using Electric Cell-Substrate Impedance Sensing Technology. Biosensors 2019, 9, 56.

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