Investigating the Importance of Physiological Cell Culture Conditions in Modeling Cancer Metabolism and Metabolic Reprogramming
dc.contributor.author | Gardner, Georgina | |
dc.date.accessioned | 2024-05-23T17:56:00Z | |
dc.date.available | 2024-05-23T17:56:00Z | |
dc.identifier.uri | http://hdl.handle.net/10464/18410 | |
dc.description.abstract | Standard cell culture conditions do not mimic the physiological environment of cancer cells. Traditional culture media contain metabolites at concentrations that far exceed conditions measured in vivo, and oxygen is often unregulated, exposing cells to atmospheric oxygen concentrations (~18%), rather than the 0-3% O2 measured in solid tumours in vivo. Recently, plasma-like media have been developed to address these limitations, aiming to improve culture conditions and maintain biologically relevant cancer phenotypes in vitro. However, these conditions remain unrepresentative of interstitial fluid in solid tumours. The goal of this thesis was to investigate how physiological culture conditions affect cancer cell behaviours, specifically cell metabolism and adaptive metabolic responses. In the first data chapter of this thesis, I explored nutrient exhaustion in physiological plasma-like medium (Plasmax) at 18% and 5% O2, and the adaptive mechanisms by which cancer cells can maintain survival under metabolic stress conditions. Here, I found that glucose and amino acid depletion from Plasmax over 48 hours is associated with several adaptive mechanisms consistent with metabolic reprogramming in vivo. Given these responses, I hypothesized that a media formulation designed using metabolite concentrations from tumour interstitial fluid may modulate metabolic phenotypes in a similar manner, providing a more physiologically relevant culture model for cancerous cells. Data chapter 2 addresses this hypothesis, whereby I formulated a novel cell culture medium using quantitative metabolite data from murine pancreatic ductal adenocarcinoma (PDAC) tumour interstitial fluid, named Tumour Microenvironment Medium (TMEM), and investigated the effects of TMEM and 1.5% O2 on an adapted murine PDAC cell line (KPCY). Importantly, I found that tumour-like conditions elicited a substantial transcriptional and functional response in cultured cells, modulating cell proliferation, migration, glucose utilization, and mitochondrial bioenergetics in ways relevant to in vivo cancer biology. Overall, the results of this thesis highlight the responsiveness of cultured cells to their environment, and the importance of representative culture conditions in the acquisition of biologically accurate experimental data. | en_US |
dc.language.iso | eng | en_US |
dc.publisher | Brock University | en_US |
dc.subject | Cell culture | en_US |
dc.subject | Cancer | en_US |
dc.subject | Physiological | en_US |
dc.subject | Metabolism | en_US |
dc.title | Investigating the Importance of Physiological Cell Culture Conditions in Modeling Cancer Metabolism and Metabolic Reprogramming | en_US |
dc.type | Electronic Thesis or Dissertation | en_US |
dc.degree.name | M.Sc. Biological Sciences | en_US |
dc.degree.level | Masters | en_US |
dc.contributor.department | Department of Biological Sciences | en_US |
dc.degree.discipline | Faculty of Mathematics and Science | en_US |
refterms.dateFOA | 2024-05-23T17:56:01Z |