Development of microfluidic systems for studying functional connectivity between in vitro neuronal co-cultures

• Main Author: Robertson, Graham
• Published: University of Strathclyde 2015
• Subjects: 610.28
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.668879

The brain is a fascinating machine that is fundamental to our existence as conscious individuals. This is highlighted during neurological disorders that can have a devastating impact on the sufferer's aptitude and quality of life. There is much which is not yet understood about what bhappens during neurological disorders including the changes which occur at a cellular level that affect synaptic communication between neurons. One method of studying these synaptic connections and how they change during disorders is through in vitro neuronal cell cultures which are a valuable tool for investigating cellular mechanisms. Recently, microfluidic techniques have enabled new methods of patterning cells in vitro and can provide precise control of the extracellular environment. Compartmentalised devices have been created that allow for certain characteristics of neurological disorders to be modelled in vitro. However, current methods of applying drugs to neuronal network in such devices are often performed manually which can limit their value as it is impractical to switch between multiple solutions. In this thesis, a method is initially developed for quantifying the synaptic comunication that occurs between functionally connected neural networks that are held in isolated environments. This was investigated using primary hippocampal neurons grown in a compartmentalised device. One sub-network of neurons was chemically stimulated while both presynaptic and postsynaptic responses were observedsimultaneously using Ca²⁺ imaging. Additionally, to address the currently limited methods of altering the extracellular environemtns in neuronal microfluidic devices, a microfluidic perfusion system was developed that can switch between multiple solutions. This was applied to compartmentalised neural networks while their cellular activity was monitored using Ca²⁺ imaging. Overall, the methods developed here can be used to study neurological mechanisms in a controlled manner and have the potential to be used in the screening of novel drugs and therapeutics targeted at neurodegenerative disorders.