Developing microelectrode and microfluidic devices for studying neuronal development
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In this thesis technologies are developed to make integrated devices to study neural activity in vitro: microelectrode arrays (MEAs), microfluidic modules and human neural stem cells (hNSCs). My aim was to develop novel in vitro models for neurodegenerative diseases such as multiple sclerosis (MS). As MS is a chronic neuroinflammatory disease that results in loss of myelin it evokes changes in neuronal conduction velocity in the central nervous system (CNS). Therefore, developing a system that could electrically follow the progress of myelination through immunocytochemistry and via conduction velocity would be of great value in MS research. As a step towards this goal, I developed and fabricated functional custom MEAs on which the electrical activity of human dopaminergic neurons (differentiated from hNSCs) and mouse spinal cord cells was recorded. Microfluidic microchannels measuring 5 μm wide were successful in separating the cell bodies of human cerebral cortical neurons (hCCNs) from the axons in two different compartments. Mouse spinal cord cultures were electrically active from 2 days in vitro (DIV) and remained active up until 52 DIV. Nominal conduction velocity (NCV) measurements were recorded from these cultures on commercial MEAs from 6 to 24 DIV. NCV increased from 0.03 m/s at 2 DIV to 15.00 m/s at 24 DIV indicating increasing myelination. Combining this data with molecular methodologies promises new approaches in developing more human-relevant models for MS research and will provide a deeper understanding of the process of myelination and possible new treatments that may one day cure MS.