Concurrent shear stress and chemical stimulation of mechano-sensitive cells by discontinuous dielectrophoresis
Concurrent shear stress and chemical stimulation of mechano-sensitive cells by discontinuous dielectrophoresis
Microfluidic platforms enable a variety of physical or chemical stimulation of single or multiple cells to be examined and monitored in real-time. To date, intracellular calcium signalling research is, however, predominantly focused on observing the response of cells to a single mode of stimulation; consequently, the sensitising/desensitising of cell responses under concurrent stimuli is not well studied. In this paper, we provide an extended Discontinuous Dielectrophoresis procedure to investigate the sensitising of chemical stimulation, over an extensive range of shear stress, up to 63 dyn/cm2, which encompasses shear stresses experienced in the arterial and venus systems (10 to 60 dyn/cm2). Furthermore, the TRPV4-selective agonist GSK1016790A, a form of chemical stimulation, did not influence the ability of the cells' to remain immobilised under high levels of shear stress; thus, enabling us to investigate shear stress stimulation on agonism. Our experiments revealed that shear stress sensitises GSK1016790A-evoked intracellular calcium signalling of cells in a shear-stimulus dependent manner, as observed through a reduction in the cellular response time and an increase in the pharmacological efficacy. Consequently, suggesting that the role of TRPV4 may be underestimated in endothelial cells-which experience high levels of shear stress. This study highlights the importance of conducting studies at high levels of shear stress. Additionally, our approach will be valuable for examining the effect of high levels of shear on different cell types under different conditions, as presented here for agonist activation.
Soffe, Rebecca
c1670df9-4ac4-4abc-8c48-3f6048a4e9fb
Baratchi, Sara
aec99b88-f6af-499d-b71d-2cfbc5fab461
Tang, Shi Yang
1d0f15c6-2a3e-4bad-a3d8-fc267db93ed4
Mitchell, Arnan
33a18604-17e3-421e-b895-0f420d332cd4
McIntyre, Peter
c05d25d3-e22e-4561-a0c5-c6bb14c24955
Khoshmanesh, Khashayar
72608a1e-0e41-4360-8065-98d872493aed
4 April 2016
Soffe, Rebecca
c1670df9-4ac4-4abc-8c48-3f6048a4e9fb
Baratchi, Sara
aec99b88-f6af-499d-b71d-2cfbc5fab461
Tang, Shi Yang
1d0f15c6-2a3e-4bad-a3d8-fc267db93ed4
Mitchell, Arnan
33a18604-17e3-421e-b895-0f420d332cd4
McIntyre, Peter
c05d25d3-e22e-4561-a0c5-c6bb14c24955
Khoshmanesh, Khashayar
72608a1e-0e41-4360-8065-98d872493aed
Soffe, Rebecca, Baratchi, Sara, Tang, Shi Yang, Mitchell, Arnan, McIntyre, Peter and Khoshmanesh, Khashayar
(2016)
Concurrent shear stress and chemical stimulation of mechano-sensitive cells by discontinuous dielectrophoresis.
Biomicrofluidics, 10 (2), [024117].
(doi:10.1063/1.4945309).
Abstract
Microfluidic platforms enable a variety of physical or chemical stimulation of single or multiple cells to be examined and monitored in real-time. To date, intracellular calcium signalling research is, however, predominantly focused on observing the response of cells to a single mode of stimulation; consequently, the sensitising/desensitising of cell responses under concurrent stimuli is not well studied. In this paper, we provide an extended Discontinuous Dielectrophoresis procedure to investigate the sensitising of chemical stimulation, over an extensive range of shear stress, up to 63 dyn/cm2, which encompasses shear stresses experienced in the arterial and venus systems (10 to 60 dyn/cm2). Furthermore, the TRPV4-selective agonist GSK1016790A, a form of chemical stimulation, did not influence the ability of the cells' to remain immobilised under high levels of shear stress; thus, enabling us to investigate shear stress stimulation on agonism. Our experiments revealed that shear stress sensitises GSK1016790A-evoked intracellular calcium signalling of cells in a shear-stimulus dependent manner, as observed through a reduction in the cellular response time and an increase in the pharmacological efficacy. Consequently, suggesting that the role of TRPV4 may be underestimated in endothelial cells-which experience high levels of shear stress. This study highlights the importance of conducting studies at high levels of shear stress. Additionally, our approach will be valuable for examining the effect of high levels of shear on different cell types under different conditions, as presented here for agonist activation.
This record has no associated files available for download.
More information
Accepted/In Press date: 17 March 2016
Published date: 4 April 2016
Identifiers
Local EPrints ID: 481730
URI: http://eprints.soton.ac.uk/id/eprint/481730
ISSN: 1932-1058
PURE UUID: acf086a0-3f54-4005-aa9c-42f325970853
Catalogue record
Date deposited: 06 Sep 2023 16:53
Last modified: 18 Mar 2024 04:13
Export record
Altmetrics
Contributors
Author:
Rebecca Soffe
Author:
Sara Baratchi
Author:
Shi Yang Tang
Author:
Arnan Mitchell
Author:
Peter McIntyre
Author:
Khashayar Khoshmanesh
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics