Enhancing high-throughput neurophysiology research through multiwell, multielectrode array capability
Lead Research Organisation:
Keele University
Department Name: Faculty of Natural Sciences
Abstract
Diseases and injuries in the brain, spinal cord or associated nervous system cells have some of the most severe implications for patients. Coupled with this, there is a significant cost to healthcare systems in treating these disorders. There are currently no cures, meaning this is a critical clinical goal for research.
Research into these diseases is often based on isolated cultures of cells. These cultures are where cells are extracted from the target organ and grown in a lab in a dish. This allows scientists to mimic injury and disease processes in simpler systems than found in the body. Specific injury processes can be easily observed and any positive impact of therapy also measured.
Currently, assessment of the models is largely achieved using microscopy and visualisation of cell behaviour. Whilst this is a powerful technique, some cells (for example those found in the brain and spinal cord) carry out their functions by electrical communication. This electrical communication is often disturbed or lost in disease and injuries so careful monitoring of it can provide important information relating to the disease/injury.
The funding we are requesting will allow us to grow cells in a dish as normal, but by using a new system - a multielectrode array - we will also be able to measure the electrical communication in the cell cultures. This will be key when studying diseases of electrically active cells and researching whether new therapies can restore the electrical communication required for function.
Research into these diseases is often based on isolated cultures of cells. These cultures are where cells are extracted from the target organ and grown in a lab in a dish. This allows scientists to mimic injury and disease processes in simpler systems than found in the body. Specific injury processes can be easily observed and any positive impact of therapy also measured.
Currently, assessment of the models is largely achieved using microscopy and visualisation of cell behaviour. Whilst this is a powerful technique, some cells (for example those found in the brain and spinal cord) carry out their functions by electrical communication. This electrical communication is often disturbed or lost in disease and injuries so careful monitoring of it can provide important information relating to the disease/injury.
The funding we are requesting will allow us to grow cells in a dish as normal, but by using a new system - a multielectrode array - we will also be able to measure the electrical communication in the cell cultures. This will be key when studying diseases of electrically active cells and researching whether new therapies can restore the electrical communication required for function.
Technical Summary
Scientists in the Keele Centre for Regenerative Medicine Research employ a range of cell/tissue culture models for MRC related projects including (i) developing traumatic neurological injury models; (ii) interfacing medical bioelectronics with neural tissue and (iii) establishing models of neurodegenerative and neurodevelopmental disorders. Traditionally, experiments are analysed by immunocytochemistry to provide cell phenotype, morphology and protein expression level data. However, a key functional measure of cell activity is the electrophysiological profile. Current electrophysiological apparatus in Keele relies on single cell patch clamp and one-well multielectrode array (MEA) systems. These approaches require significant technical expertise and are low throughput. Further, the current MEA system is over 10 years old with a difficult user-interface and cannot run multiple experiments in parallel.
To address these issues we request funding to purchase a Maestro Multiwell MEA from Axion Biosystems. The device allows medium to high-throughput recording of each well of a culture plate (6, 12, 24 and 96 well plates) meaning controls and numerous experimental conditions can be analysed simultaneously. The system is equipped with a fully controllable environment chamber for maintaining desired cell culture conditions (normoxia/hypoxia can be achieved) for long term recordings. The software is user friendly and flexible for multiple experimental paradigms including measuring action potential and network electrophysiology but also monitoring cell proliferation, cytotoxicity, cell adhesion, cell migration, wound healing, and barrier function (for example for blood brain barrier investigations). Electrical stimulation can also be applied to any electrode in any well for regenerative stimulation experiments or pacing for cardiomyocyte maturation. We are also requesting a Lumos light stimulator for coupling with optogenetic experiments, to enhance the system functionality.
To address these issues we request funding to purchase a Maestro Multiwell MEA from Axion Biosystems. The device allows medium to high-throughput recording of each well of a culture plate (6, 12, 24 and 96 well plates) meaning controls and numerous experimental conditions can be analysed simultaneously. The system is equipped with a fully controllable environment chamber for maintaining desired cell culture conditions (normoxia/hypoxia can be achieved) for long term recordings. The software is user friendly and flexible for multiple experimental paradigms including measuring action potential and network electrophysiology but also monitoring cell proliferation, cytotoxicity, cell adhesion, cell migration, wound healing, and barrier function (for example for blood brain barrier investigations). Electrical stimulation can also be applied to any electrode in any well for regenerative stimulation experiments or pacing for cardiomyocyte maturation. We are also requesting a Lumos light stimulator for coupling with optogenetic experiments, to enhance the system functionality.
