Spatially patterned vascular cell co-cultures for combined electrical-optical monitoring of cell-cell communication
Lead Research Organisation:
University of Strathclyde
Department Name: Biomedical Engineering
Abstract
Overview
Local communication between biological cells, through direct cell-cell interactions and paracrine signalling, is central to normal tissue physiology. Disruption of this communication plays a central role in numerous disease processes, including cardiovascular disease. This project will develop a completely new approach to dynamically monitoring vascular cell communication, by combining microfabrication techniques and surface functionalisation to create an impedance analysis microdevice with spatial cell patterning capability. This will provide novel mechanistic insights into cardiovascular disease that will open up opportunities for the development of improved treatments.
Background
Cardiovascular disease results from a complex remodelling of the blood vessel wall. Two key aspects of this process are initial endothelial cell (EC) dysfunction and subsequent phenotypic change and accumulation of smooth muscle cells (SMCs), with inherent disruption in local cell-cell communication. EC damage and SMC proliferation also underlie restenosis, which remains the Achilles heel of stent treatments. The processes driving these changes remain poorly understood, greatly hampering efforts to develop new treatments. Novel systems that mimic cardiovascular remodelling are therefore required both to tease apart the complex signalling mechanisms involved and for drug screening models.
Dr Sandison and Dr McCormick's combined research experience leaves them well placed to address this need. Dr Sandison's recent publications have focussed on detailed investigations into phenotypic changes in SMCs, including developing methods for isolating, characterising and imaging vascular populations. Dr McCormick has developed a fully automated impedance spectroscopy system for monitoring cell responses in near real-time, showing that EC and SMC co-cultures give rise to distinct impedance spectra that are strongly influenced by cell-cell communication mechanisms. Together with Dr Sandison's broad experience in microsystems technology, this research base will ensure successful delivery of the project.
Proposed Research
A novel device incorporating transparent, thin-film, microfabricated electrodes, enabling simultaneously electrical-optical cell monitoring, will be developed. By correlating time-lapse imaging with impedance measurements, it will enable detailed characterisation of communication between cell populations, providing a series of validated impedance profiles for different co-culture environments. A combination of silane chemistry and electrochemical desorption will be employed to sequentially create adherent regions for patterning different cell types upon specific microelectrode regions. Significantly, modes of cellular communication will be teased apart by systematically modifying microelectrode geometries. To better mimic in vivo conditions, controlled shear stress will be applied by integration into a microfluidic system.
Objectives:
(1) Produce a new impedance analysis microdevice, developing protocols for sequential cellular patterning of multiple cell types.
(2) Acquire control spectra for single vascular populations (e.g. ECs, SMCs) as they proliferate towards confluency.
(3) Perform impedance analysis of spatially patterned co-cultures with both large (biochemical communication between cells only) and small (enabling physical communication via cellular processes) gaps between different populations.
(4) Develop an integrated microfluidic system for analysis under continuous/pulsatile flow and for controlled drug delivery, examining the effect of drugs used in stent coatings.
As well as producing a well-characterised in vitro model of vascular cell-cell communication, the resulting microsystem will be suitable for upscaling into an array format for multiplexed, label-free drug screening.
Local communication between biological cells, through direct cell-cell interactions and paracrine signalling, is central to normal tissue physiology. Disruption of this communication plays a central role in numerous disease processes, including cardiovascular disease. This project will develop a completely new approach to dynamically monitoring vascular cell communication, by combining microfabrication techniques and surface functionalisation to create an impedance analysis microdevice with spatial cell patterning capability. This will provide novel mechanistic insights into cardiovascular disease that will open up opportunities for the development of improved treatments.
Background
Cardiovascular disease results from a complex remodelling of the blood vessel wall. Two key aspects of this process are initial endothelial cell (EC) dysfunction and subsequent phenotypic change and accumulation of smooth muscle cells (SMCs), with inherent disruption in local cell-cell communication. EC damage and SMC proliferation also underlie restenosis, which remains the Achilles heel of stent treatments. The processes driving these changes remain poorly understood, greatly hampering efforts to develop new treatments. Novel systems that mimic cardiovascular remodelling are therefore required both to tease apart the complex signalling mechanisms involved and for drug screening models.
Dr Sandison and Dr McCormick's combined research experience leaves them well placed to address this need. Dr Sandison's recent publications have focussed on detailed investigations into phenotypic changes in SMCs, including developing methods for isolating, characterising and imaging vascular populations. Dr McCormick has developed a fully automated impedance spectroscopy system for monitoring cell responses in near real-time, showing that EC and SMC co-cultures give rise to distinct impedance spectra that are strongly influenced by cell-cell communication mechanisms. Together with Dr Sandison's broad experience in microsystems technology, this research base will ensure successful delivery of the project.
Proposed Research
A novel device incorporating transparent, thin-film, microfabricated electrodes, enabling simultaneously electrical-optical cell monitoring, will be developed. By correlating time-lapse imaging with impedance measurements, it will enable detailed characterisation of communication between cell populations, providing a series of validated impedance profiles for different co-culture environments. A combination of silane chemistry and electrochemical desorption will be employed to sequentially create adherent regions for patterning different cell types upon specific microelectrode regions. Significantly, modes of cellular communication will be teased apart by systematically modifying microelectrode geometries. To better mimic in vivo conditions, controlled shear stress will be applied by integration into a microfluidic system.
Objectives:
(1) Produce a new impedance analysis microdevice, developing protocols for sequential cellular patterning of multiple cell types.
(2) Acquire control spectra for single vascular populations (e.g. ECs, SMCs) as they proliferate towards confluency.
(3) Perform impedance analysis of spatially patterned co-cultures with both large (biochemical communication between cells only) and small (enabling physical communication via cellular processes) gaps between different populations.
(4) Develop an integrated microfluidic system for analysis under continuous/pulsatile flow and for controlled drug delivery, examining the effect of drugs used in stent coatings.
As well as producing a well-characterised in vitro model of vascular cell-cell communication, the resulting microsystem will be suitable for upscaling into an array format for multiplexed, label-free drug screening.
