Development of Microfluidic Systems for Studying Functional Connectivity Between In Vitro Neuronal Co-Cultures

Graham Robertson

Research output: ThesisDoctoral Thesis

Abstract

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 happens 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 communication 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 observed simultaneously using Ca2+ imaging. Additionally, to address the currently limited methods of altering the extracellular environments 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.
LanguageEnglish
QualificationPhD
Supervisors/Advisors
  • Zagnoni, Michele, Supervisor
  • Bushell, Trevor, Supervisor
Place of PublicationGlasgow
Publisher
Publication statusPublished - 11 Sep 2015

Fingerprint

Microfluidics
Coculture Techniques
Neurons
Nervous System Diseases
Switches
Imaging techniques
Cellular neural networks
Equipment and Supplies
Communication
Cell culture
Brain
Screening
Preclinical Drug Evaluations
Neural networks
Neurodegenerative Diseases
Cell Culture Techniques
Perfusion
Quality of Life
In Vitro Techniques
Pharmaceutical Preparations

Keywords

  • neurological disorders
  • synaptic communication
  • in vitro neuronal cell culture
  • microfluidic techniques
  • neurodegenerative disorders

Cite this

@phdthesis{4cee71868fb1401b9159acc902d39151,
title = "Development of Microfluidic Systems for Studying Functional Connectivity Between In Vitro Neuronal Co-Cultures",
abstract = "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 happens 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 communication 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 observed simultaneously using Ca2+ imaging. Additionally, to address the currently limited methods of altering the extracellular environments 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.",
keywords = "neurological disorders, synaptic communication, in vitro neuronal cell culture, microfluidic techniques, neurodegenerative disorders",
author = "Graham Robertson",
year = "2015",
month = "9",
day = "11",
language = "English",
publisher = "University of Strathclyde",

}

Development of Microfluidic Systems for Studying Functional Connectivity Between In Vitro Neuronal Co-Cultures. / Robertson, Graham.

Glasgow : University of Strathclyde, 2015. 200 p.

Research output: ThesisDoctoral Thesis

TY - THES

T1 - Development of Microfluidic Systems for Studying Functional Connectivity Between In Vitro Neuronal Co-Cultures

AU - Robertson, Graham

PY - 2015/9/11

Y1 - 2015/9/11

N2 - 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 happens 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 communication 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 observed simultaneously using Ca2+ imaging. Additionally, to address the currently limited methods of altering the extracellular environments 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.

AB - 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 happens 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 communication 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 observed simultaneously using Ca2+ imaging. Additionally, to address the currently limited methods of altering the extracellular environments 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.

KW - neurological disorders

KW - synaptic communication

KW - in vitro neuronal cell culture

KW - microfluidic techniques

KW - neurodegenerative disorders

M3 - Doctoral Thesis

PB - University of Strathclyde

CY - Glasgow

ER -