Thesis

Isolation and characterisation of small extracellular vesicles from the human pancreatic cancer cell line BxPC-3.

Creator
Rights statement
Awarding institution
  • University of Strathclyde
Date of award
  • 2024
Thesis identifier
  • T17095
Person Identifier (Local)
  • 202262702
Qualification Level
Qualification Name
Department, School or Faculty
Abstract
  • Extracellular vesicles come in various shapes and sizes and are released by most cell types. They have myriad roles in intercellular signalling in both physiological and pathological environments, carrying a range of lipids, proteins and nucleic acids. Their cargo is then unloaded at the target site inducing a change in their target cell. Cancers use these vesicles to their advantage for a wide range of outcomes such as immune evasion and chemoresistance leading to the reduced effect of chemotherapies and unfavourable patient outcomes. Pancreatic cancer has one of the worst outcomes of any cancer with surgery being the only cure. As surgery is only available in a small number of cases, targeted delivery of cargos directly to the tumour site is of high importance to efficiently target and destroy cancer cells with high effectiveness without the toxic off-target effects of chemotherapy drugs. Hijacking the bodies postal system has gained interest in the last decade for the delivery of therapeutic drugs. The low immunogenicity and inherent biocompatibility of extracellular vesicles avoids a hurdle of other nanoparticles such as the toxicity of metals. Various techniques for loading and functionalising extracellular vesicles have evolved to the extent of clinical trials. However, these therapies are yet to make it to market. This highlights the importance of finding a standardised method for isolating extracellular vesicles on a large scale with few contaminants. Ultracentrifugation is the gold standard for the isolation of extracellular vesicles. However, a growing list of techniques are becoming available, potentially giving more reproducible and higher yield and purity outcome. In addition to the growing arsenal of isolation techniques, characterisation is becoming equally more in-depth allowing for the rigorous analysis of both the extracellular vesicles and the techniques themselves. The International Society of Extracellular Vesicles has released 3 mission statements to aid researchers in standardising their techniques allowing for reliable comparison in 2014, 2018 and 2023. In this work 3 widely available techniques were compared using the human Pancreatic cancer cell line BxPC-3. The techniques were chosen for their cost-effectiveness and ability to be scaled up to manufacturing scales.
Advisor / supervisor
  • Hoskins, Clare
Resource Type
DOI
Embargo Note
  • The digital version of this thesis is restricted to Strathclyde users only until 23rd September 2029.

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